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AMNUAl'REPORT^ 



MMX P'ROGRAIVI.ACT!VITIES ■^■: 
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S„ DEPARTMEMT OF HEALTH AND HUMAN SERVICES 
tiblic Healtia Service National Institutes of Health 




ANNUAL REPORT, 

OF 

PROGRAM ACTIVITIES 

NATIONAL CANCER INSTITUTE (^OS.j 

Fiscal Year 1981 

Part II-A 

Division of Cancer Biology and Diagnosis 



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Page 



DIVISION OF CANCER BIOLOGY AND DIAGNOSIS 

ANNUAL REPORT 
October 1, 1980 through September 30, 1981 

TABLE OF CONTENTS 

EXTRAMURAL RESEARCH PROGRAM 

ASSOCIATE DIRECTOR 

Summary Report '- 

Tumor Biology Program 

Summary Report 5 

Grants 21 

Contract 51 

Immunology Program 

Summary Report 5 J 

Grants 65 

Contracts "-' 

Breast Cancer Program Coordination Branch 

Summary Report ^'■■'- 

Grants ^25 

Contracts ^^^ 

Cancer Diagnosis Research Program 

Summary Report ^^'- 

Grants 201 

Contracts 209 

Biology Support Contracts 253 



EXTRAMURAL RESEARCH PROGRAM 
DIVISION OF CANCER BIOLOGY AND DIAGNOSIS 

The Extramural Research Program covers four broad scientific areas: Tumor 
Biology, Immunology, Cancer Diagnosis and Breast Cancer Program. The first two 
are primarily grant based efforts, the latter programs employ both contracts and 
grants in supporting the sceintists at large. 

Three years ago, an effort was started to convert then primarily contract based 
program to one oriented on ROl and POl grants. In this year we witness a 
successful achievement of this goal. With exception of the Cancer Diagnosis 
Program, where clinical evaluations and instrument developments will continue to 
be supported by contracts, the remaining programs will concentrate almost ex- 
clusively on grants. 

Another change in the management of the DCBD Extramural Research Program concerns 
the Advisory Committees, which have slowly been deactivated during the past year. 
At the end of this year only the Breast Cancer Task Force will continue as an 
active body advising the staff of this organ site program. On the other hand, 
the Board of Scientific Counselors will enlarge its responsibilities to cover the 
entire division. It will review the program annually, provide concept review on 
new projects and act as site visit team to intramural laboratories and branches. 
Senior staff review for relevance will be carried out as before by the Steering 
Committee consisting of NCI personnel. 

The extramural research budget this year reached $138,000,000 of which 
$126,000,000 were in grant supported projects and $12,000,000 in contracts 
(including this Division's share of the FCRC effort). 

The Tumor Biology Program supports a broad spectrum of basic biological and 
biochemical research in the pursuit of one goal, defining properties of cancer 
cells and tumors that uniquely distinguish them from normal, healthy cells and 
tissues. If we could define these properties and learn how to manipulate the 
biological signals or modify biochemical reactions responsible for the aberrant 
behavior of cancer cells, applied methods could be developed for the successful 
diagnosis and therapy of cancer patients. The three areas of investigation which 
correspond to different theories of how to control the development and pro- 
gression of neoplastic disease include understanding the basic biochemical mech- 
anisms involved in growth control, studies of the changes that occur at the 
molecular level which lead to cancer cell invasion and accession of detailed 
biological and biochemical information about the processes which induce cancer 
cell differentiation. If the genetic program of an actively growing cancer could 
be changed to one of terminal differentiation, then the malignant tumor could be 
rendered harmless. The above emphasis of the Tumor Biology Program in the areas 
of growth, invasion and differentiation provides a convenient and purposeful way 
of viewing the role of basic biological research with the ultimate goal of curing 
cancer . 

The scope of the Tumor Biology Program has changed considerably over the last 
five years. Mostly, this has been due to research advances in our knowledge of 



gene expression and growth requirements of eukaryotic cells, as well as technical 
advancements in somatic cell genetics, somatic cell hybridization and DNA recom- 
binant biology. The Tumor Biology Program has become increasingly focused on the 
properties of cancer cells rather than on the cell biology of eukaryotic cells. 

Immunology Program of the National Cancer Institute supports studies which con- 
tribute to an understanding of the role of the immune system on the development, 
growth and spread of tumors. In line with organizational changes within the 
Institute, the emphasis for the Immunology Program is now centered upon mecha- 
nistic studies with responsibility for the more applied immunological efforts in 
detection and diagnosis residing in the Diagnosis Program and responsibility for 
explicit treatment studies involving immunologic manipulation residing in the 
programs of the Division of Cancer Treatment. 

Some of the more obvious areas supported by the Immunology Program include 
studies in: molecular genetics and biology of immunoglobulins, natural cell 
mediated immunity, monoclonal antibodies and immune response genes. Following 
conferences and meetings were supported by the Program: "Mechanisms in Human 
Cancer Immunology Symposion", "Host Defense in Neoplasia" and "Mechanisms in 
Cell Mediated Cytoxicity". To illustrate the program more specifically, the 
following activities were supported: the synthesis, structure and mechanism of 
action of antibodies capable of reacting with tumor cells and of humoral factors 
other than antibody which participate, activate and regulate the immune response 
to tumors: complement, interferon, lymphokines, lymphoid cell growth factors, 
helper factors and suppressor factors. Other studies covered were the immuno- 
biology of Ijmiphocytes , monocytes and macrophages that participate in anti-tumor 
responses including their development, heterogeneity, interactions and mech- 
anisms of action. 

Considerable effort was devoted to the identification, isolation and characteri- 
zation of cell surface determinants which serve as target antigens for the immune 
response to tumors and of cell surface determinants of Ijrmphocytes and macro- 
phages which are involved in actual responses of these cells to tumors. Immune 
surveillance against the development of tumors of various origins by all immune 
mechanisms included studies on T cell immunity, macrophage reactivity, natural 
killer cell activity, etc. 

The Cancer Diagnosis Research Program emphasizes research in early detection, 
diagnosis, tumor localization, and monitoring of the disease. The major objec- 
tive of the Program is to recognize or detect cancer at the earliest possible 
stage to allow appropriate therapy which should improve the chances for the 
control of cancer. 

The Diagnosis Research Program consists of projects in eight disciplinary cate- 
gories: biochemistry, immunodiagnosis , cytology, pathology, radiological im- 
aging, non-radiological imaging, nuclear medicine and multiple disciplinary 
projects. Biomedical methods of diagnosis and detection involve a variety of 
substances such as hormones, enzymes, proteins and metabolic products, surface 
characteristics of tumor cells and their chemical characterization. Immuno- 
diagnosis projects can be subdivided into those dealing with circulating tumor 
antigens or markers, such as oncofetal antigens, hormones, enzymes, and glyco- 
proteins; projects dealing with tumor associated antigens, studies of lympho- 
cytes and projects dealing with antibodies to tumors. 



Diagnostic cytology research projects include the development and testing in- 
strumentation and cell markers that can be used to differentiate normal and 
atypical cells. Imaging studies in this program use conventional x-ray approach 
with the goal of reducing the exposure doses without compromising the quality of 
imgages. Another arm involves methods other than x-ray and radionuclides; such 
as the use of proton and heavy ion beams, nuclear magnetic resonance (NMR) , 
ultrasound, and thermography. Finally, Diagnosis Program supports the research 
on screening of lung, colon and endometrial cancers. 

Breast Cancer Program promotes and supports mul tidisciplinary research projects 
that will lead to improved methods of diagnosis, prognosis, treatment and pre- 
vention of breast cancer. This is the oldest organ site program and the only one 
managed by NCI personnel exclusively. To help direct the program, advise from 
the Breast Cancer Task Force Committee is obtained, whose members suggest areas 
in need of emphasis, innovative research ideas and workshop topics aimed at 
exploring rapidly developing areas. The following topics exemplify the field 
coverage: "Luteal Phase Defects and Breast Cancer Risk", "Clonogenic Assays and 
Chemotherapy Sensitivity", "Chemical Carcinogen-Hormone Interaction in Trans- 
formation of Mammary Epithelial Cells In Vitro " , "Monoclonal Antibodies in 
Breast Cancer", and "Diet and Breast Cancer Risk . 

The staff of the Branch organizes requests for investigator-initiated appli- 
cations (RFA's) or Program Announcements (PA's), and requests for contract pro- 
posals (RFP's) depending upon the mechanism of funding considered. In addition, 
there exists an information section responsible for monthly production of the 
publication Intercom which provides up-to-date listings of scientific papers on 
breast cancer research in biology, epidemiology, diagnosis, and treatment; a 
list of meetings and conferences related to the disease; and abstracts of pres- 
entations made at workshops. The publication is sent to investigators and 
institutions throughout the world. Another publication of the Intracom provides 
abstracts or summaries of the published literature as well as those on grant and 
contract activites. 



TUMOR BIOLOGY PROGRAM 
Description 

The Tumor Biology Program supports a broad spectrum of basic biological 
and biochemical research (See Appendix I) in the pursuit of one over- 
riding goal, defining properties of cancer cells and tumors that uniquely 
distinguish them from normal, healthy cells and tissues. The supposition 
is that if we can define these properties and learn how to manipulate or 
change the biological signals and/or modify biochemical reactions respon- 
sible for the aberrant behavior of cancer cells, applied methods can be 
developed for the successful treatment, diagnosis and therapy of cancer 
patients. Within this goal are three major areas of investigation which 
correspond to different theories of how to control the development and 
progression of neoplastic disease. The first is understanding the basic 
biochemical mechanisms involved in growth control, whether these involve 
particular external signals that initiate the process of cell division or 
particular internal molecules more directly responsible for DNA replica- 
tion and metabolism. This kind of information can lead to the develop- 
ment of specific hormonal and drug therapies. The second is studying 
changes that occur at the molecular level which lead to cancer cell 
invasion . The invasive behavior of cancer cells is a prerequisite to 
malignancy, or the ability of tumors to invade surrounding tissues, 
escape normal host defense mechanisms and become established at multiple 
secondary metastatic sites. Theoretically, if the invasive properties 
of malignant tumors can be controlled and these tumors confined to parti- 
cular sites, metastasis, the major killer in cancer patients, will not 
occur. The third is to develop detailed biological and biochemical 
information about the processes which induce cancer cell differentiation . 
There is good reason to believe that many kinds of cancers will respond 
to external stimuli and differentiate. If the genetic program of an 
actively growing cancer could be changed to one of terminal differenti- 
ation, then the malignant tumor could be rendered harmless. Although 
the above emphasis of the Tumor Biology Program in the areas of growth, 
invasion and differentiation is stated in simple terms, it provides a 
convenient and purposeful way of viewing the role of basic biological 
research to the ultimate goal of curing cancer. 

The kinds of information developed in the Tumor Biology Program provide a 
foundation for and relate directly or indirectly to nearly every other 
program area within the National Cancer Institute. The importance of 
basic tumor biology research to the National Cancer Plan is reflected by 
the large $50 million commitment of the NCI to this program area in FY 
1981 (See Table I). Appendix II provides a complete listing of all 
grants supported by the Tumor Biology Program. 



Selected Scientific Developments 

A. Tumor Cell Invasiveness: 

The most lethal property of malignant cells is their ability to metastasize 
to distant sites. However, before metastases can be established, the 
malignant cell must invade the surrounding tissues. Because an understanding 
of the invasive process remains a central problem for tumor biology, the 
Tumor Biology Program organized and sponsored a "Tumor Cell Invasion Work- 
shop." Three areas were chosen for discussion: (1) Methods and Models for 
Studying Tumor Cell Invasion; (2) Tumor Cell Penetration of the Extracellular 
Matrix; and (3) The Role of Cell Motility in the Invasive Process. The 
objectives of the workshop were to bring a small group of scientists together 
for a day and a half to explore the state-of-the-art in these three areas 
and, through an emphasis on discussion rather than formal presentation, to 
define the limitations of current experimental approaches and to suggest new 
approaches required to advance our knowledge of the field. The participants 
were carefully chosen from both the U.S. and foreign countries for their 
current research interest in problems related to invasive and metastatic 
processes and for their diversity of training from biochemistry to embryologj 
to pathology. This was a unique group of individuals (See Appendix III). 

Several major issues were brought to the group's attention before any 
substantive discussion of the three major topics was initiated. First, there 
was a recognized problem between the basic scientists and the pathologists in 
the use of the term malignant. It was generally agreed that it is not 
appropriate to use in vitro criteria alone to call a tumor cell malignant; 
thus, viral transformation or any other kind of transformation should not 
be referred to as malignant transformation unless this could be confirmed by 
accepted _in vivo criteria. No one could agree upon whether malignant should 
refer to a tumor that kills or a tumor that metastasizes or a tumor that 
invades the surrounding tissue or all three. Unfortunately, some kinds of 
tumors (e.g. basal cell carcinomas and primary tumors of the brain) kill the 
patient but do not invade or metastasize. There is no doubt that an accept- 
able definition is required for malignancy that satisfies both clinical sit- 
uations and experimental systems if contradictions and confusions in the 
scientific literature are to be avoided in the future . Furthermore, invasion 
is currently defined very crudely by what a pathologist sees in a microscope. 
Everyone agreed that a rigorous definition of invasion is not possible until 
the invasive process can be measured (i.e. there are clear measurable markers ). 
The second issue discussed was more a reminder to the group that invasiveness 
is not a unique property of tumor cells . During embryonic development, many 
different kinds of cells show periods of invasive behavior; in post-natal 
life, melanocytes, capillary endothelial cells and several classes of white 
blood cells retain their invasive properties. This is an important point ( 
because tumors have never been shown to exhibit any behavior or property 
that is completely tumor-specific. This is a basic reason why the cancerous 



process is so difficult to deal with both experimentally and clinically. 
The third issue which captured the entire groups attention and which offered 
the greatest challenges and implications for experimental analysis of the 
invasive phenotype, was phenotypic heterogenity of cell populations within 
a tumor. George Poste (CA 30192) presented startling information that 
cloned B16 mouse melanoma cell lines, selected for their increased metastatic 
potential or low metastatic potential, formed tumors that were very unstable; 
the population of cells within a tumor apparently derived from a single 
clone, generated phenotype heterogeneity with respect to invasive properties 
or drug sensitivity very quickly. Subsequent to Poste 's presentation, this 
observation was surprisingly confirmed by Fidler using a different cloned 
melanoma system, by Heppner (CA 27419) using a variety of cloned mouse mammary 
tumor cell lines and by Varani (CA 29550) using cloned mouse fibrosarcoma 
cells. It was postulated that the stability of any given tumor cell phenotype 
depended upon the interaction or presence of other phenotypes; clearly, 
one cloned tumor phenotype is not likely to persist in an rn vivo situation. 
Thus, the problem of heterogeneity has very serious implications that must 
be dealt with experimentally if we are to learn more about the invasive 
phenotype . 

Ideally, model systems for studying the invasive process should be (1) amen- 
able to a combination of in vitro and in vivo techniques; (2) permit quanti- 
tation of invasion and; (3) allow for recovery of invasive cells for further 
study. There are no model systems that fulfill all these criteria, but 
there are a number of useful approaches for studying invasion that have been 
developed within the last few years. Poste (CA 30192) has developed a system 
whereby tumor cells are measured for their ability to penetrate various natural 
membranes and/or membrane-cell-matrix structures; this technique offers great 
promise because the matrix that is invaded can be varied and cells recovered 
on the basis of their relative invasive properties. Peter Jones presented a 
culture system in which smooth muscle cells are layered on a tissue culture 
dish followed by endothelial cells; a clear basal lamina forms between these 
two kinds of cells, which closely resembles the in vivo situation. When human 
fibrosarcoma cells were layered on this cell matrix, they penetrated readily 
and this penetration was accompanied by proteolytic activity. This model 
allows for quantitation of release of tissue proteins as well as histological 
correlates of the invasive process. Pauli's system (Pauli et al. , 1980) used 
carcinogen induced bladder carcinomas and had the advantage that it correlated 
in vitro and in vivo studies well. X-ray analysis _in vivo could demonstrate 
invasion into baby rat bone; cell lines iii vitro derived from the same bladder 
carcinoma could be studied for their invasion of cartilage. Reich presented 
a very interesting system for measuring invasion; human cancer cells were 
layered on the allantoic membrane of a fertilized chicken egg. The lungs of 
the chicken embryo were measured for human plasminogen activator as the 
measure of invasion. There were other model systems briefly discussed but 
these were not new to the field. All model systems require an appropriate 
cell as well as an appropriate stroma that is to be invaded. Although signi- 
ficant progress has been made in developing new model systems, a major problem 
is that the cells used have not had their biological properties adequately 
defined. This is especially important to consider if we are to assume that 
phenotypic heterogeneity rather than stability is the more likely situa- 
tion encountered in vivo. 



Penetration of the extracellular matrix, by tumor cells is fundamental to our( 
understanding of the invasive proocess. This section of the workshop was 
perhaps the most interesting in terms of new information provided. The most 
striking information was provided by Kuettner (CA 21566) in his description 
of recent information about the complex structure of the extracellular matrix. 
(Jhile only a few years ago, the extracellular matrix was believed to consist 
primarily of collagen, now it has been established that there are many differ- 
ent genotypic collagens, tissue-specific proteoglycans, laminin and fibronec- 
tion. The latter two components have been the subject of intense investiga- 
tions because of their apparent involvement in cell-cell interactions. Fur- 
thermore, the matrix is highly underhydrated , which would lead one to believe 
that molecules such as nutrients and proteins might have considerable difficulty 
passing through the matrix. However, Pietro Gullino presented information 
that the matrix (i.e. interstitial space) does exchange fluids, nutrients and 
proteins very rapidly with the vascular system; thus, the matrix is a very 
dynamic structure. Many of the participants demonstrated that there are 
specific degradative enzymes for each component within the matrix, and there 
was good reason to believe that each degradative enzyme could be inhibited by 
a specific inhibitor. This explains why past research efforts to correlate 
one kind of collagenase activity with tumor cell invasiveness have been 
unsuccessful. There are many enzymatic activities and inhibitory activities 
to consider. Furthermore, the development of tumor heterogeneity can create 
microenvironments which stimulate the production of key enzymes and/or inhibi- 
tors that initiate the invasive process. Although the participants in the 
workshop favored the hypothesis that the degradative activity of enzymes 
produced by the tumor or the host are actively involved in the invasive ( 
process, they could not rule out a simpler physical process. However, all 
of the discussion pointed toward a very active system in which the environment 
could trigger many different enzymatic or inhibitory activities. Everyone 
agreed that further research should carefully define the nature of the matrix 
being studied and the specificity of the enzyme being studied, as well as 
determine whether natural inhibitors of the enzyme were present under differ- 
ent conditions. 

The motility of the tumor cell is essential if it is to move from the primary 
tumor and reach the vascular and lymphatic systems. Because there has been 
a considerable emphasis in the literature which describes the fibriullar net- 
works within tumor cells that may be responsible for cell motility, this 
area was not discussed thoroughly. No compelling evidence was presented to 
support the hypothesis that tumor cells are intrinsically more motile than 
their corresponding normal cells. Ward (Orr _et_ al. , 1979); Wass et al. , 
1980) discussed the possibility that tumor cells respond to chemotactic 
factors , supporting the possibility that tumor cell invasion is an active, 
directional process. He believed that his information supported a model in 
which tumor cells moved similarly to leukocytes in response to factors. 
This is an interesting theory because tumor cell exodus from the primary 
site has often been observed to follow leukocytes through the same channels. 
Furthermore, capillary growth toward tumors, as discussed in the next section, 
may follow the path of infiltrating leukocytes. There could be some interest- 
ing common denominators relating tumor cell invasion, capillary invasion, 
and leukocyte motility. ( 



Varani (CA 29950), using mouse fibrosarcoma cells, showed data which supported 
non-directed motility as another component of tumor cell migration. By altering 
the surface components responsible for cell adhesion, a clear correlation could 
be made between decreased cell motility, increased cell adhesion in vitro 
and tumor takes after sub-cutaneous injection iji vivo . 

To summarize, better model systems are critical to experimental analysis of the 
invasive process. Invasion of tissue matrices by both normal and tumor cells 
appears to involve a combination of tissue destruction (i.e. degradative enzymes 
and inhibitors of degradative enzymes) and directional movement (i.e. chemotactic 
factors). The microenvironment of the host and tumor heterogeneity are likely 
to be critical factors in understanding what controls the invasive process. 

B. Angiogenesis 

The importance of tumor vascularization to tumor growth has been recognized 
for many years. Most solid malignant tumors have the capacity to induce the 
generation of new capillary growth to meet their ever increasing requirement 
of blood supply. The phenomenon, called angiogenesis, is a topic of great 
interest to the Tumor Biology Program. Certain recent developments from the 
projects supported through the program have encouraged the suggestion that 
control of tumor growth through therapy with antiangiogenetic factors may be 
possible. Three processes appear to contribute to the overall process of 
capillary growth: 1) degradation of basement membrane of the blood vessel in 
the area of a developing outgrowth; 2) endothelial cell migration and; 3) 
endothelial cell replication. Sophisticated systems have been designed to 
investigate each of these processes. 

Folkman et al.(1971), first described a Tumor Angiogenesis Factor (TAF) that 
promoted the invasion of an avascular area by capillaries. Production of 
sufficient quantities of the factor (or factors) to allow purification and 
characterization has, however, been an extremely difficult problem. For 
years only complicated _in vivo assays were 'available to test for the factor 
and the results were only semi-quantitative. 

In 1976, Fenselau and Mello described an _in vitro assay for TAF. Homogenates 
of Walker 256 carcinoma stimulated proliferation of endothelial cells prepared 
from fetal bovine aorta. A big breakthrough came in 1979 when Folkman and 
associates reported development of a cloned long-term culture of bovine 
capillary endothelial cells, the growth of which required the presence of 
media pre-conditioned by incubation with cells from sarcoma 180 tumors. The 
tumor-derived angiogenesis factors not only induced proliferation of the 
capillary endothelial cells but migration as well. This migration could be 
quantitated and was proportional to the concentration of tumor-conditioned 
medium present (Zetter ,1980). These in vitro systems, then, seem to support 
what has been observed Jji vivo , that neovascularization involves the migration 
of endothelial cells from pre-existing vessels towards the source of the 
angiogenesis factor and that there is a directional elongation of growth of 
new capillary sprouts. These same cloned endothelial cells under the proper 
culture conditions, with the tumor-derived factors, will organize into capil- 
lary tubes, form branches and assemble an entire capillary network. This 
appears to be the first demonstration of _in vitro angiogenesis (Folkman and 
Haudenschild, 1980). 

9 



Although attempts to characterize TAF have still not provided definitive \ 
information about molecular structure, Fenselau et al. (1981) now report 
the preparation of purified material from the Walker 256 rat tumor with a 
molecular weight of less than 800 daltons. The factor is mitogenic for 
endothelial cells and induces neovascularization in an ±n_ vivo assay. 

Another recent report (Azizkhan et al. , 1980), Indicates that a product of 
mast cells (a type of white blood cell), probably heparin, also stimulates 
the migration of the cultured bovine capillary endothelial cells without 
inducing proliferation. Since the accumlation of mast cells at a tumor site 
precedes the arrival of new blood vessels, the mast cell product may somehow 
direct the construction of the new vasculature. 

The third process of neovascularization, the fragmentation of the existing 
basement membrane of the blood vessel at the point of new capillary growth, 
and the ability of these new sprouts to invade surrounding tissues, suggests 
the activation of proteolytic activity. Rifkin and associates (Moscatelli et 
al. , 1980) have been able to demonstrate the production of collagenase from 
cultured human endothelial cells by stimulation with the tumor promoter 12-0- 
tetradecanoyl phorbol-13-acetate. Thus, one more response to angiogenic stim- 
uli may be the increased secretion of collagenase. 

Although little is known about the factors that contribute to angiogenesls , it 
has been possible to inhibit their activity. Unfortunately, the inhibitory 
factors have been as elusive as the stimulatory factors. , One approach 
has been the preparation of specific protease and collagenase inhibitors ( 
that prevent vascular invasion. These are found in a variety of cell types. 
Another has focused on extraction products of cartilage, a unique tissue in 
that it normally is devoid of any vasculature. The natural resistance of 
cartilage to tumor invasion has been well documented (Kuettner and Pauli, 1981) 
and also appears to be due in part to endogenous protease inhibitors. In in 
vivo assay systems, invading capillaries moving towards a tumor stimulus grow 
around but cannot invade implanted pieces of cartilage. Extracts of cartilage 
inhibited vascular growth when infused into live animals bearing tumor implants 
in their eyes (Langer et al. , 1980). The extract had no effect on cultured 
cells of the same tumor type. In the culture system of capillary endothelial 
cells, migration induced by tumor-derived factors is also inhibited by the 
presence of leukocyte interferon (Brouty-Boye and Zetter, 1980). 

Prevention of angiogenesls could have an extraordinary effect on cancer treat- 
ment. The inhibitors so far identified are all natural products so that side 
effects are minimal. Furthermore, the effects seem to be specific for endothe- 
lial cells — they prevent new vascularization only. The next few years should 
produce some exciting new developments. 

C. Teratocarcinoraa 

Some intriguing and potentially useful new developments in research with the 
teratocarcinoma cell system also warrant special attention in this report. 
Tumors, derived from germ cells, which develop in the ovary or testis of mice 
and humans, may be of either of two types. Teratocarcinomas are malignant ( 



10 



timiors which grow from embryonal carcinoma cells themselves derived from germ 
cells. These embryonal carcinoma cells are "stem" cells which may continue to 
proliferate or may differentiate into chaotic arrangements of somatic cells 
typical to an embryo. If all the cells differentiate the tumors become benign 
and are known as teratomas. Embryonal carcinoma cells from the teratocarcinomas 
have been established rn vitro and provide the special model system now used 
extensively in research. Certain of the cell lines (PSA-1, for example) retain 
their totipotent character and will produce a variety of differentiated cell 
types in vitro or in vivo , while some variants (F9, for example) have become 
nullipotent and will not differentiate spontaneously, but simply keep replicat- 
ing. 

The development of transplantable teratocarcinomas in mice can be attributed 
to the work of Leroy Stevens (1980). He has defined both genetic and 
environmental factors involved in the tumorigenesis and supplied the transplant- 
able teratocarcinomas to the rest of the research community. However, it was 
Pierce and co-workers (1967) who first formulated a theory, prompted by this 
cell system, that differentiation of stem cells within a tumor might deplete 
the malignant element and result in a benign neoplasm. The implication of this 
was a new approach to cancer therapy. 

The demonstration (Mintz et al. , 1975), that microinjection of mouse teratocar- 
cinoma cells into blastocysts from normal mice and subsequent implantation 
into the uteri of pseudopregnant females resulted in the formation of normal 
chimeric mice, was noteworthy for its suggestion that malignancy was not the 
result of a special gene content but rather of the regulation of genes. The 
mechanism whereby the 80-cell-blastocyst can shut down the "malignancy" of 
embryonal carcinoma cells is unknown, however, there is a limit to the number 
of carcinoma cells they can tolerate. Injection of 20 malignant cells into 
a normal blastocyst results in the birth of chimeric offspring bearing tumors 
(Papaioannou, 1975). In fact, it was recently shown (Pierce et al. , 1979) 
that control of malignancy is lost when the blastocyst contains more then 
two embryonal carcinoma cells. When three or more cells are injected into 
the blastocyst the hybrid cell mass forms tumors typical of embryonal carinoma 
in the peritoneum of mice. 

One of the best studied teratocarcinoma cell lines, the F9, does not undergo 
spontaneous differentiation although it can be induced to differentiate by 
treatment with retinoic acid and other compounds. Further the F9 cells, when 
infected with SV40 virus do not support replication of the virus nor express 
the typical virus-specific antigens. After retinoic acid induction of the cells, 
the antigens are expressed. Analagous results have been attained with a more 
sophisticated F9 clone transformed with a plasmid containing SV40 genes linked 
to a herpes simplex virus thymidine kinase. The cell line expresses the thymidine 
kinase but does not express the adjacent viral antigens unless treated with 
retinoic acid (Linnenback et al. , 1980) 

The majority of teratocarcinoma cell culture systems maintain their pluripotency 
and become benign differentiated cell types in vitro , with limited lifespans. 
However when cultures are initiated with a 1:1 mixture of pluripotent (PSA-1) and 
nullipotent (the F9) cells, the environment restricts differentiation of the 

11 



( 

PSA-1 cells and they begin to die. No detectable inhibitor from the F9 cells 
has yet been found (Rosenstraus and Levine, 1979). Hybrid cells prepared by 
fusion of pluripotent with nullipotent teratocarcinoma cells have been studied 
independently by two different groups to determine which characteristic is 
dominant. Rosenstraus _et al (1980) reported that such hybrids, when injected 
into mice, developed tumors with the characteristics of the pluripotent cells, 
i.e. , they had a spectrum of differentiated tissue. Oshima et al (1981), in 
contrast, prepared similar hybrids and observed the growth of tumors that 
were 95 - 100% embryonal carcinoma stem cells. 

Another related research project which exploits the teratocarcinoma system is 
that of Martin. She is attempting to explain the inactivation of one of the 
X chromosomes in the female mouse zygote that occurs at about day four during 
normal embryological development. Certain of the embryonal carcinoma cell 
lines from ovarian tumors have two active X chromosomes and during differ- 
entiation one of these is inactivated, so the dissection of the related events 
may be possible (Martin, et al. , 1978). 

The Tumor Biology Program has begun to formulate an outline for a workshop on 
teratocarcinoma to be held during the next fiscal year. A survey of selected 
investigators throughout the country indicates that this would be an ideal time 
to organize such a conference. Investigators currently in this arena are 
embryologists, biologists, pathologists and immunologists but molecular 
biologists and geneticists and biochemists, all with their special techniques, 
must also be encouraged to get involved, perhaps through collaborations. It ( 
is also an important time to delineate both the limitations and the potentials 
of the teratocarcinoma system. retinolc acid (Linnenback et al. , 1980). 

D. A Unifying Hypothesis for Neoplastic Transformation 

A major frustration of tumor biology has been that each kind of cancer 
appears to be a different disease. Comparative studies of normal and 
cancer cells always show rather striking differences, but these differences 
are never exactly the same for each kind of cancer. An intriguing 
set of hypotheses has been postulated that may unify many apparently disparate 
observations of the past and present. During the past decade, protein 
phosphorylation reactions (protein kinase activity) have been found to 
function in the regulation of a large number of cellular processes. In 
fact, a unique class of protein kinases appears to participate in malignant 
transformation (Langan, 1980). The best studied system is the transformation 
of cells by Rous sarcoma virus. The transforming protein appears to be a 
protein kinase (src protein kinase); in addition, cells uninfected by the 
virus appear to produce a very similar if not identical protein but at a 
much lower level. Although this observation is interesting in itself, Racker 
and his colleagues have recently confirmed that the enzyme Na^/K"^ ATPase 
which controls the sodium-potassium balance of the cell is not only inefficient 
in cancer cells but that its activity is controlled by a cascade of different 
protein kinases. Surprisingly, the src protein kinase and the first kinase in 

( 



12 



the cascade of events that activates the Na+Zr*" ATP'ase crossreact immunologi- 
cally and are similar by other structural criteria. Thus, this is the first 
time that a transforming gene product has been linked to the disruption of a 
specific process in the cell. Racker postulates that these protein kinases 
may trigger a whole assortment of biochemical changes leading to various 
neoplastic phenotypes. Even more speculative is that oncogenic viruses 
originate from movable elements within the genome (i.e. transposons)and have 
carried away proto-oncogenes (e.g. protein kinases) which are normally present 
in the cellular DNA (Fox, 1981). Coincidentally , it has been postulated that 
human cancers arise more frequently from genetic transpositions rather than 
from conventional mutational events (Cairns, 1981). Proto-oncogenes in the 
normal genome are not just speculation because several laboratories have now 
shown that host genes involved in neoplastic transformation can be detected 
by DNA-raediated gene transfer experiments (Rigby, 1981). Thus, a more unifying 
hypothesis for neoplastic transformation might include the following: 

(1) Many normal cells contain proto-oncogenes. 

(2) Transforming viruses not only contain genes derived from host 
proto-oncogenes but also originate from the movable genetic elements 
within the cell (i.e. transposons). 

(3) Transpositions of proto-oncogenes which allow for increased 
genetic expression result in the neoplastic phenotype. 

(4) Proto-oncogenes are often protein kinases. 

Obviously, the above model is highly speculative and many questions remain 
unanswered and untested, but the very real possibility that there may be some 
general simplicity attributed to many transformation processes is intriguing 
and exciting. 

E. Tumor Markers 

One of the common threads interwoven throughout the literature of cancer 
biology is a wide-ranging abnormality of gene expression (Weinhouse, 
1980). One reason why the early diagnosis of cancer has improved is because 
different kinds of cancers typically produce enzymes or hormones that 
are not normally associated with the tissue of origin. Unfortunately, 
these so-called "tumor markers" have never been identified as specific 
qualitative differences between a normal and cancer cell, but always 
seem to represent quantitative differences due to abnormalities of gene 
regulation that result in a different programming of protein synthesis. 
Tumor markers may, however, provide important clues about the initiation 
and progression of malignant disease. Several investigators have docu- 
mented the presence of altered glycoproteins on the surface of trans- 
formed and malignant cells. These have prompted speculation that mem- 
brane-bound glycoproteins may mediate the abnormal behavior of cancer 
cells. These studies also have stimulated a search for soluble glyco- 
protein tumor markers in biological fluids which are a product of some- 
kind of "shedding" process. Several years ago a cancer-associated galactosyl 
transferase isoenzyme was reported. This study has been carried further, 



13 



r 

and it has now been established that in the sera of patients with local- 
ized neoplasia, there is also a distinctive glycopeptide that functions 
as an acceptor for the galactosyl transferase isoenzyme. Paradoxically, 
this glycopeptide inhibits tumor growth in tissue culture and in animal 
models. However, one speculation is that it represents a host defense 
product that is produced too late and in insufficient concentrations to 
effect a clinical remission (Podolsky and Isselbacher, 1980). It is 
hoped that a further understanding of this glycopeptide' s role in affect- 
ing the natural growth of malignant cells will not only establish its 
value as a diagnostic indicator but also as a biological response modi- 
fier. Another laboratory has reported the presence of a specific glyco- 
protein with a molecular weight of 50,000-55,000 which is significantly 
elevated in the sera from cancer patients with localized or metastatic 
malignancies (Bolmer and Davidson, 1981). The ectopic production of 
glycoprotein hormones and their subunits is regularly reported in cancer 
patients (e.g. Blackman et_ al, 1980). With the development of better 
methods of purifying and detecting glycoproteins (e.g. monoclonal anti- 
bodies) there is every reason to believe that we will learn a great deal 
more about the significance of glycoprotein cancer markers in tumorigen- 
esis. 

F. Malignancy as a Dominant or Recessive Trait 

For the last five years there has been considerable controversy over whether/""^ 
the transformed phenotype _in vitro and tumorigenicity _in vivo are expressed ffb 
recessive or dominant traits in animals cells. To help answer this question 
somatic cell hybridization techniques have been used in which normal and 
tumorigenic cells are fused and the resulting hybrids are tested for normal 
and transformed properties, as well as tumorigenicity. Unfortunately, 
despite intensive efforts to determine the recessiveness or dominance of 
malignant traits, the question remains confused. In nearly every case, the 
expression of cell transformation and tumorigenicity is dominant in viral- 
transformed cells. But in the case of non-virally transformed cells of either 
rodent or human origin, the answer is not clear; some results with somatic cell 
hybrids demonstrate dominance while others suggest recessiveness. The emergence 
of chromosome-mediated and DNA-mediated gene transfer techniques should be 
valuable tools to test whether the results obtained by somatic hybridization 

are correct. We are now at the stage where it should be possible to clone 
some of the cancer genes using the DNA mediated transfer approach and recom- 
binant DNA technologies. The availability of cloned mammalian cancer genes 
should allow a detailed study of their structure and organization and of the 
regulation of their expression. Furthermore, the possibility of introducing 
entire cancer chromosomes or cloned cancer genes into pluripotent teratocar- 
cinoma stem cells (see above) should provide valuable insights about their 
regulation during cell differentiation and development (Croce, 1980). 



14 





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15 



APPENDIX I ^ 

TUMOR BIOLOGY REFERRAL GUIDELINES 

The Tumor Biology Program supports a broad spectrum of research with animal 
and human cell systems which includes fundamental and comparative studies in 
histology, pathology, cell biology, developmental biology, molecular biology, 
embryology, biochemistry, and genetics. The goal of this program is to 
develop a better understanding of the biological processes in tumors and tumor 
cells that are responsible for uncontrolled growth and invasion of surrounding 
tissues. The majority of studies are aimed at determining characteristic 
differences in structure, function, biochemical composition and molecular 
mechanisms between normal and neoplastic cells or between progressive states 
of tumor development and differentiation. Areas supported include: 

- Tumor Progression (e.g., angiogenesis , neovascularization, invasiveness, 

metastasis) 

- Differentiation and Neoplasia (e.g., teratomas, teratocarcinomas , 

hepatomas, neuroblastomas, melanomas. Friend leukemias) 

- Genetics of Neoplasia (e.g., karyotypes, somatic cell genetics, somatic 

cell hybrids, gene mapping, inherited genetic syndromes) 

- Normal, Abnormal and Neoplastic States of Growth (e.g., cell growth 

cycle, fast growing vs slow growing) 

- Normal and Neoplastic Cell Behavior (e.g., cell-cell adhesions, cell 

movement and migration-microtubules-microf ilaments , cell-cell 
communication) ^ 

- Membrane Synthesis, Structure, Composition and Function (e.g., trans- V 

port, receptors, sugar transferases, glycolipids, glycoproteins, 
basement membrane, extracellular matrix, gap junctions, tight 
junctions) 

- Cell Metabolism - Regulation of (e.g., energy metabolism, metabolic 

pathways, enzymes, cGMP , cAMP) 

- Nucleic Acid and Protein Synthesis and Processing, Interactions and 

Modifications - Regulation of (e.g., DNA, rRNA, sRNA, mRNA, 
regulation of gene expression, histones, nonhistones, chromatin) 

- Nutritional, Hormonal and Protein Factor Requirements for Tumor 

Growth, Maintenance and Differentiation (e.g., cellular factors, 
serum factors, peptide hormones, steroid hormones) 

- Model Systems for Studying Tumor Growth, Invasiveness and Metastasis 

EXCEPTIONS AND QUALIFICATIONS: 

Excluded from this program are all research studies focused on breast cancer 
detection and diagnosis and immunology. Also excluded are those studies 
oriented primarily to cancer therapy. 

Experimental studies with prokaryotic and plant systems are supported only 
when these studies are clearly defended in terms of potential knowledge that 
will enhance cancer research. Except for comparative studies of normal and 
virally transformed cells in culture, this program is not concerned with the 
occurrence, nature or mechanism of action of virally induced cancers. 



16 



PARTICIPANTS 



FOR THE WORKSHOP ON "TUMOR CELL INVASION" 



Dr. Peter Armstrong 
University of California 
Davis, California 

Dr. Chitra Biswas 
Massachusetts General Hospital 
Boston, Mass. 

Dr. Guenter W. Albrecht-Buehler 
Cold Spring Harbor Laboratory 
Cold Spring, New York 

Dr. Bill R. Brinkley 
Baylor College of Medicine 
Houston, Texas 

Dr. Carlo M. Croce 
Wistar Institute 
Philadelphia, PA 

Dr. Carol Erickson 
University of California 
Davis, California 



Dr. Isaiah J. Fidler 

Frederick Cancer Research Center 

Frederick, Maryland 

Dr. Judah Folkman 

Children's Hospital Medical Center 

Boston, Mass. 

Dr. Howard Gershman 

Case Western Reserve University 

Cleveland, Ohio 

Dr. Pietro Gullino 
National Cancer Institute 
Bethesda, Maryland 

Dr. Ian Hart 

Frederick Cancer Research Center 

Frederick, Maryland 



17 



r 

REFERENCES 



Azizkhan, R. , Azizkhan, J. , Zetter, B. and Folkman, J. , "Mast Cell Heparin 
Stimulates Migration of Capillary Endothelial Cells in vitro " , J. Exp. Med . 
152: 931-944, 1980. (CA 14019) 

Blackman, M.R. , Weintraub, B.D. , Rosen, S.W. , Kourides, I. A., Steinwascher, 
K. and Gail, M.H. , "Hijman Placental and Pituitary Glycoprotein Hormones 
and Their Subunits as Tumor Markers: A Quantitative Assessement , " JNCI, 
65:81-93, 1980. (CA 23185) 

Bolmer, S.D., and Davidson, E.A. , "Preparation and Properties of a Glycoprotein 
Associated with Malignancy", Biochemistry , in press, 1981. (CA 15483) 

Brouty-Boye, D. and Zetter, B. , "Inhibition of- Cell Motility by Interferon", 
Science 208:516-518, 1980. (CA 28540) 

Cairns, J., "The Origin of Human Cancers", Nature , 289: 353-357, 1981. 

Croce, CM., "Cancer Genes in Cell Hybrids", Biochim . et Biophys. 
Acta , 605:411-430, 1980. (CA 20741, CA 23568) 

Fenselau, A. and Mello, R.J. , "Growth Stimulation of Cultured Endothelial 
Cells by Tumor Cell Homogenates" , Cancer Res . 36:3269-3273, 1976. (CA 15381^ 

Fenselau, A., Watt, S. and Mello, R.J. , "Tumor Angiogenic Factor: Purification 
from the Walker 256 Rat Tumor", J. Biol. Chem . , in press, 1981. (CA 15381) 

Folkman, J., Haudenschild, C.C. and Zetter, B. , "Long-term Culture of 
Capillary Endothelial Cells", Proc . Natl . Acad . Sci . , U.S.A., 76:5217-5221, 
1979. (CA 14019) 

Folkman, J. and Haudenschild, C. , "Angiogenesis in vitro ". Nature 288:551- 
556, 1980. (CA 14019) 

Folkman, J., Merler, E. , Abernathy, C. and Williams, G. , "Isolation of a Tumor 
Factor Responsible for Angiogenesis", J. Exp. Med . 133:275-288, 1971. (CA 14019) 

Fox, J.L. , "Cancer Researchers Identifying Common Clues", Chemical and 
Engineering News , pp. 16-20, March 16, 1981. (CA 08964, CA 14454 & CA 15823) 

Kuettner, K.E. and Pauli, B.U. , "Resistance of Cartilage to Invasion" in Bone 
Metastasis , eds. Gilbert, H.A. , Weiss, L. and Monsen, D.C.G. (G.K. Hall & Co., 
Boston), pp. 131-165, 1981. (CA 21566) 

Langan, T. , "Malignant Transformation and Protein Phosphorylation", Nature 
286: 329-330, 1981. (CA 12877) 

Langer, R. , Conn, H. , Vacanti, J., Haudenschild, C. and Folkman, J., "Contro( f 
Tumor Growth in Animals by Infusion of an Angiogenesis Factor", Proc. Natl . Acad . 
Sci . , U.S.A., 77:4331-4335, 1980. (CA 14019) 

18 



Linnenback, A., Huebner, K. and Croce, CM., "DNA-trans formed Murine 
Teratocarcinoma Cells: Regulation of Expression of Simian Virus 40 Tumor 
Antigen in Stem Versus Differentiated Cells", Proc . Natl . Acad . Sci . , U.S.A., 
77:4875-4879, 1980. (CA 20741, CA 21069) 

Martin, G.R. , Epstein, C.J. , Travis, B. , Tucker, G. , Yatziv, S., Martin, D.W. , 
Jr., Clift, S. and Cohen, S. , "X-Chromosome Inactivation During Differentia- 
tion of Female Teratocarcinoma Stem Cells in vitro " , Nature 271:329-333, 

1978. (CA 25966) 

Mintz, B., Illmensee, K. and Gearhart, J.D. , "Developmental and Experimental 
Potentialities of Mouse Teratocarcinoma Cells from Embryoid Body Cores" in 
Teratomas and Differentiation , ,eds. Sherman, M.I. and Solter, D. , (Academic, 
N.Y.), 1975. 

Moscatelli, D. , Jaffe, E. and Rifkin, D.B., "Tetradecanoyl Phorbal Acetate 
Stimulates Latent Collagenase Production by Cultured Endothelial Cells", 
Cell 20:343-351, 1980. (CA 23753) 

Orr, W. , S.H. Phan, J. Varani, P. A. Ward, D.L. Kreutzer, R.O. Webster, P.M. 
Henson, "Chemotactic Factor for Tumor Cells Derived from the C5a Fragment of 
Complement Component C5," Proc . Natl . Acad. Sci . , U.S.A., 76:1986-1989, 

1979. (CA 29551, CA 29550) 

Oshima, R. , McKerros, J. and Cox, D. , "Murine Embryonal Carcinoma Hybrids: 
Decreased Ability to Spontaneously Differentiate as a Dominant Trait", J. 
Cell Physiol . , in press, 1981. (CA 27580) ~ 

Papaioannou, V.E., McBurney, M.W. and Gardner, R.O., "Fate of Teratocarcinoma 
Cells Injected into Early Mouse Embryos, Nature (London) 258:70-73, 1975. 

Pauli, B.U. , S.N. Anderson, V.A. Memoli, and K.E. Kuettner, "Development of 
an in Vitro and in Vivo Epithelial Tumor Model for the Study of Invasion," 
Cancer Research , 40:4571-4580, 1980. (CA 21566) 

Pierce, G.B., "Model for a Developmental Concept of Cancer", Current Topics 
Develop . Biol . 2:223-246, 1967. (CA 15823) 

Pierce, G.B., Lewis, S.H. , Miller, G. J. , Moritz, E. and Miller, P., "Tumori- 
genicity of Embryonal Carcinoma as an Assay to Study Control of Malignancy 
by the Murine Blastocyst", Proc . Natl . Acad. Sci . , U.S.A., 76:6649-6651, 1979. 
(CA 15823) 

Podolsky, D.K. and Isselbacher, K.J., "Cancer-Associated Galactosyltransferase 
Acceptor", Cancer , 45: 1212-1217, 1980. (CA 14294) 

Rigby, P.W.J. , "The Detection of Cellular Transforming Genes", Nature 
290: 186-187, 1981. (CA 28946) 

Rosenstraus, M.J. , Balint, R.F. and Levine, A.J. , "Pluripotency of Somatic Cell 
Hybrids Between Nullipotent and Pluripotent Embryonal Carcinoma Cells", Somatic 
Cell Genetics 6:555-565, 1980. (CA 26014, CA 28106) 

19 



K 

Rosenstraus, M.J. and Levine, A.J. , "Alterations in the Developmental Potential! 
of Embryonal Carcinoma Cells in Mixed Aggregates of Nullipotent and Pluripotent 
Cells", Cell 17:337-346, 1979. (CA 26014, CA 28106) 

Stevens, L.D. , "Genetic Influences on Teratocarcinogenesis in Mice" in A Century , 
of Mammalian Genetics and Cancer , View in Midpassage , ed. Russell, E.S.7 
(Alan R. Liss, Inc., N.Y.), pp. 1929-2029, in press, 1981 . (CA 02662)' 

Wass, J. A., J. Varani, P. A. Ward, "Size Increase Induced in Walker Ascites Cells i 
by Chemotactic Factors," Cancer LeHers , 9:313-318, 1980. (CA 29551, CA 29550) 

Weinhouse, S. , "New Dimensions in the Biology of Cancer", Cancer, 45:2975- 
2980, 1980. (CA 10916) . 

Zetter, B.R. , "Migration of Capillary Endothelial Cells is Stimulated by Tumour- 
derived Factors", Nature 285:41-43, 1980. (CA 28540) 



20 



CELL SURFACE 



ROl-CA-02758 Steroid Metabolism in Tumors and Normal Tissues 
Kandutsch Jackson Laboratory 

ROl-CA-08759 Structure, Biosynthesis and Function of Glycoproteins 
Kornfeld Washington University 

ROl-CA-10917 Phosphate Transport and Metabolism in Carcinoma Cells 

Levinson University of Texas Hlth. Sci. Ctr. San Antonio 



ROl-CA-12150 
Baumann 



Unusual Lipids in Cancer Tissues 

University of Minnesota at Austin 



ROl-CA-12306 

Cunningham 



Preparation of Cancer Chemotherapy Abstracts, Volume XI 
University of California Irvine 



ROl-CA-12753 
Grimes 



Cell Surfaces and Cells Transformed by Oncogenic Viruses 
University of Arizona 



ROl-CA-12790 Substrate and Cation Transport in Human Leukocytes 
Lichtman University of Rochester 



ROl-CA-12920 

Oppenheimer 



Mechanisms of Intercellular Adhesion 

California State University Northridge 



ROl-CA-13402 Surface Membranes in Normal and Cancer Cells 
Atkinson Yeshiva University 

ROl-CA- 13605 Chemistry and Measurement of Intracellular Adhesion 
Steinberg Princeton University 



ROl-CA-14370 
Maslow 



Specificity of Normal and Cancer Cell Interactions 
Roswell Park Memorial Institute 



ROl-CA-14431 Cell Adhesion of Normal and Malignant Liver Cells 

McGuire National Jewish Hospital & Research Ctr. 



ROl-CA-14464 

Loewenstein 



A Program on the Etiology and Diagnosis of Cancer 
University of Miami 



ROl-CA-14496 
Magnuson 



Metal Ion Activation of Concanavalin A 

Washington State University 



ROl-CA-14551 Penetration of Macromolecules into Mammalian Cells 
Ryser Boston University 

ROl-CA- 14609 Molecular Basis of Cellular Adhesiveness 

Grinnell University of Texas Hlth. Sci. Ctr. Dallas 

ROl-CA-14764 Glycolipid Metabolism in Tumor and Transformed Cells 
Basu University of Notre Dame 



21 



ROl-CA-15047 

Friedberg 

ROl-CA-15483 
Davidson 

ROl-CA-16335 
Sheridan 

ROl-CA-16777 

Schutzbach 

ROl-CA-16856 

Hellerquist 

ROl-CA-17007 
Hynes 

ROl-CA-17149 
Doyle 

ROl-CA-18801 
Morre 

ROl-CA-19017 
Klebe 

ROl-CA-19130 
Warren 

ROl-CA-19144 
Buck 

ROl-CA-19158 
Biswas 

RO.l-CA-20026 
Hakomori 

ROl-CA-20421 
Krag 

ROl-CA-20424 

Goldstein 

ROl-CA-20575 
Barrett 

ROl-CA-21463 
Furcht 

ROl-CA-21923 

Baenziger 



0-Alkyl Lipids in Surface Membranes of Tumor Cells 

University of Texas Hlth, Sci. Ctr. San Antonio 

Biochemistry of Mucopolysaccharides 

Pennsylvania State Univ. Hershey Med. Ctr. 

Cell Junction Formation and Role in Cell Proliferation 

University of Minnesota of Minneapolis-St . Paul 

The Biosynthesis of Cell Envelope Glycoproteins 
University of Alabama in Birmingham 

Analysis of Eucaryotic Cell Surfaces 
Vanderbilt University 

Cell Surface Structure and Transformation 

Massachusetts Institute of Technology 

Proteins of the Hepatoma Cell Plasma Membrane 

New York State Department of Health 

Glycosphingolipid Metabolism and Tumorigenesis 
Purdue University West Lafayette 

Cell Adhesion Proteins and Malignancy 

University of Texas Med. Br. Galveston 

The Surface Membranes of Normal and Cancer Cells 

Wistar Institute of Anatomy and Biology 

Membrane Changes Caused by Tumor Virus Transformation 
Wistar Institute of Anatomy and Biology 

Mammalian Collagenase in Tumor Invasion 

Massachusetts General Hospital 

Tmmuno specif ic Glycolipids of Normal and Cancer Cells 
Fred Hutchinson Cancer Research Center 

Mutants Altered in Glycosylation of Membrane Proteins 
Johns Hopkins University 

Murine Ascites Tumor Cell Glycoproteins 
University of Michigan 

Myeloid Cell Surface Receptors: Normal and Leukemic 
University of California San Francisco 

Pathobiology of the Cell Membrane in Cancer 

University of Minnesota of Minneapolis-St. Paul 

Oligosaccharide Structure and Receptor Specificity 
Washington University 



22 



ROl-CA-22451 Contact Behavior of Developing and Transformed Cells 
Trinkaus Yale University 



ROl-CA-22659 
Chen 



Studies of Proteins Involved in Cell Interaction 
Sidney Farber Cancer Institute 



ROl-CA-22729 Hormonal Regulation of Membrane Phenotype 
Gelehrter University of Michigan 

ROl-CA-22907 Collagenase and Invasion of Head and Neck Tumors 
Huang Columbia University 

ROl-CA-23016 Animal Cell Surface Heparin Sulfate and Transformation 
Keller University of Hlth. Sci. /Chicago Med. Sch. 

ROl-CA-23095 Carcinogenesis/Membrane Enzyme and Permeability Mutants 
Hochstadt New York Medical College 



ROl-CA-23540 
Smith 

ROl-CA-23753 

Rifkin 

ROl-CA-23907 
Hakomori 



Collagen and Its Relationship to Tumors 
Boston University 

Membrane Proteins of Normal and Malignant Cells 
New York University 

Galactoprotein A and B in Normal and Transformed Cells 
Fred Hutchinson Cancer Research Center 



ROl-CA-24051 
Matta 



Synthetic Glycosides for Cancer Research 

Roswell Park Memorial Institute 



ROl-CA-24339 Membrane Lipid Asymmetry in a Tumorigenic Cell Line 
Schroeder University of Missouri Columbia 

ROl-CA-24419 Role of Glycoconjugates in Formation of Metastasis 

Skipski Sloan Kettering Institute for Cancer Res. 

ROl-CA-24488 The Controlled Initiation of Neoplasms in Drosophila 
Hanratty University of California Irvine 

ROl-CA-24553 Targeting of Liposomes to Tumor Cells 

Huang University of Tennessee Knoxville 

ROl-CA-24598 Mechanism of ^^Ga Uptake by EMT-6 Sarcoma BALB/c Mice 
Larson University of Washington 

ROl-CA-24605 Fibronectin and Its Loss in Malignant Transformation 
Vaheri University of Helsinki 

ROl-CA-25074 Cell Surface Alterations in Malignancy 

Weiser State University of New York at Buffalo 



ROl-CA-25532 

Schwa rting 



Glycolipids of Normal and Transformed Mouse Lymphocytes 

Eunice Kennedy Shriver Ctr. Mtl. Retardation 



23 



ROl-CA-25730 Membrane Proteins and Lipids in Mammary Carcinoma 
Steiner University of Kentucky 

ROl-CA-25898 Analysis of Gl in Mammalian Cells 
Baserga Temple University 

ROl-CA-26055 Structure-Function Relationships of Antitumor Lectins 
Fowler University of North Carolina Chapel Hill 

ROl-CA-26103 Plasma Membrane Synthesis in Friend Leukemic Cells 

Hunt University of Mississippi Medical Center 

ROl-CA-26122 Effect of Hormones on Cell Surface Architecture 
Baumann New York State Department of Health 

ROl-CA-26294 Glycoproteins of Normal/Malignant Human Blood Cells 
Gahmberg University of Helsinki 

ROl-CA-26814 Purine Transport by Cancer Cell Line Membrane Vesicles 
Hochstadt New York Medical College 

ROl-CA-27062 Immunobiology of Membrane: Serum Protein Interactions 
Galbraith Medical University of South Carolina 

ROl-CA-27285 Plasma Membranes and Control of Cell Growth 
Peterson Boston University 

ROl-CA-27389 Cell Surface Fibrous Proteins and Control of Metastasis 
Singer Institute for Medical Res. of Bennington 

ROl-CA-27397 Plasma Membrane Alterations in Transformed Cells 

Fink University of Colorado Hlth. Sciences Ctr. 

ROl-CA-27441 Cell Surface Changes in Epidermal Differentiation 

Brysk University of Texas Med. Br. Galveston 

ROl-CA-27455 Fibroblast Surface Antigen 

Ruoslahti La Jolla Cancer Research Foundation 



ROl-CA-27460 

Ruoslahti 



Alpha- Fetoprotein: Structure and Function 

La Jolla Cancer Research Foundation 



ROl-CA-27648 Tumorigenesis and a Cell Surface Growth Inhibitor 
Johnson Kansas State University 

ROl-CA-27655 Gene Amplification in Carcinogenesis 
Miller Columbia University 

ROl-CA-27755 Fibronectin: Proteoglycan Binding in Adhesion Sites 
Gulp Case Western Reserve University 

ROl-CA-28101 Glycosaminoglycans in Normal and Malignant Cells 
Ruoslahti City of Hope National Medical Center 



24 



ROl-CA-28287 Driving Forces for Nutrient Transport in Tumor Cells 

Smith University of Texas Hlth. Sci. Ctr. San Antonio 

ROl-CA-28548 Developing Inmunological Probes for Gap Junctions 

Johnson University of Minnesota of Minneapolis-St . Paul 

ROl-CA-28685 Phosphorylation Events and Transformed Cell Membranes 

Gordon University of Colorado Hlth. Sciences Ctr. 

ROl-CA-28867 Cell Surface Studies of Metastatic Melanoma 

Nicolson University of Texas System Cancer Center 



R23-CA-29172 
Carter 



Structure and Function of the 170K Glycoprotein 

Fred Hutchinson Cancer Research Center 



ROl-CA-29271 Pathobiology of Pancreatic Acinar Cell Carcinoma 
Rao University of Pittsburgh 

R23-CA-29814 Noncytolytic Extraction of Tumor Antigens with Butanol 

Le Grue University of Texas Hlth. Sci. Ctr. Houston 

ROl-CA-29995 Pathobiology of Malignant Cell Basement Membrane 

Furcht University of Minnesota of Minneapolis-St. Paul 

R23-CA-30030 Sodium Ion Fluxes and Mitogenic Signaling 

Smith University of Alabama in Birmingham 

ROl-CA-30117 Epithelial-Mesenchymal Interaction in Endocrine Tissues 
Reid Yeshiva University 

ROl-CA-30231 Murine T Lymphocyte Cell Surface Antigens 
Springer Harvard University 

ROl-CA-30289 Human Epithelial Cells-Extracellular Matrix Interactions 
Vlodavsky Hebrew University of Jerusalem 

ROl-CA-30381 Study of the Plasma Membrane Matrix of Lymphoid Cells 
Mescher Harvard University 



ROl-CA-30538 
Jamieson 



Interactions of Platelets and Tumors Cells 
American National Red Cross 



ROl-CA-30645 Glycosylation Mutants of Animals Cells 
Stanley Yeshiva University 



R23-CA-31004 
Barnes 



Cell-Substratum Interactions 

University of Pittsburgh 



ROl-CA-31103 Molecular Determinants of Multicellular Organization 
Hixson University of Texas System Cancer Center 

ROl-CA-31182 Ecto 5 '-Nucleotidase as a Cell Surface Reporter 
Carraway University of Miami 



25 



ROl-CA-31761 
Taub 



Cell Surface Sugars in Pathogenesis and Therapy of CML 
Columbia University 



ENZYMES 



ROl-CA-04679 
Lerner 



Biology of Normal and Malignant Melanocytes 
Yale University 



ROl-CA-10916 Metabolism of Normal and Neoplastic Tissue 
Weinhouse Temple University 



ROl-CA-11655 
Silber 



Studies of Leukocyte Metabolism 
New York University 



ROl-CA-11949 Ether Lipids in Cancer-Enzyme Mechanisms and Inhibitors 
Snyder Oak Ridge Associated Universities 



ROl-CA-12563 

Stellwagen 



Mechanism of Enzyme Induction by Cyclic Nucleotides 
University of Southern California 



ROl-CA-14881 
Shiman 



Regulation of Tyrosine Synthesis in Hepatoma Cells 

Pennsylvania State Univ. Hershey Med. Ctr. 



ROl-CA-15196 Regulation of Fatty Acid Biosynthesis in Mammary Tumors 
Ahmad Papanicolaou Cancer Research Institute 



ROl-CA-15979 

Siperstein 

ROl-CA-16184 
Felder 



Cholesterol Metabolism in Normal and Malignant Liver 
University of California San Francisco 

Regulation of Enzyme Levels in Mouse Hepatoma 

University of South Carolina at Columbia 



ROl-CA-16231 Role of Collagenases in Tumor Invasion and Metastasis 
Liener University of Minnesota at Saint Paul 



ROl-CA-21967 
Fishman 



Fundamental Enzymologic Studies Applied to Oncology 
La Jolla Cancer Research Foundation 



ROl-CA-22409 Enzyme Regulation in Normal and Neoplastic Cells 
Deuel Jewish Hospital of St. Louis 

RO1-CA-22690 Cyclic Nucleotides in Cancer and Regenerating Tissue 
Rosenberg Albany Medical College 

ROl-CA-22717 Regulation of Folate Poly-Y Glutamate Synthesis 
Shane Johns Hopkins University 

ROl-CA-23391 Cyclic GMP System in Cell Proliferation 
Shoji Emory University 

ROl-CA-23533 Tumor Immunology: Phosphatase Isoenzymes 
Doellgast Wake Forest University 



26 



ROl-CA-23743 Ornithine Decarboxylase Induction in Neoplastic Cells 
Byos University of California Riverside 

ROl-CA-24966 Catalase Isozymes in Normal and Neoplastic Tissues 
Nishimura U.S. PHS Hospital 

ROl-CA-25005 The Regulation of Mammalian Enzyme Synthesis 
Greengard Mount Sinai School of Medicine 

ROl-CA-25088 Human Pancreatic Ribonuclease and Cancer 

Glitz University of California Los Angeles 

ROl-CA-25337 Drugs and Flavoprotein Mediated Superoxide Formation 
Powis Mayo Foundation 

ROl-CA-25473 Cyclic AMP Metabolism in Cultured Epithelial Cells 
Niles Boston University 

ROl-CA-25617 The Collagenolytic System of Invasive Tumors 

Dabbous University of Tenn. Center Health Scien. 

ROl-CA-26033 White Blood Cell Function in Hematologic Disorders 
Nathan Children's Hospital Medical Center 

ROl-CA-26102 Protein Kinase System in Rapidly Growing Hepatomas 
Criss Howard University 

ROl-CA-26470 Regulation of Dihydrof olate Reductase Gene Expression 
Johnson Ohio State University 



ROl-CA-26546 

Canellakis 



Regulation of Enzyme Activity in Normal and Tumor Cells 
Yale University 



ROl-CA-27073 Cellular Enzyme Changes in Childhood Leukemia 
Dunn Virginia Commonwealth University 

ROl-CA-27491 Form and Function of Normal and Neoplastic Ferritins 
Massover College of Medicine and Dentistry of NJ 

R23-CA-27500 The Control of Cyclic GMP in Human Lymphocytes 
Takemoto Kansas State University 

ROl-CA-27572 Ribonucleotide Reductase of Tumor Cells 
Cory University of South Florida 

ROl-CA-27674 Control of Pyrimidine Biosynthesis in Mammalian Cells 
Evans Wayne State University 

ROl-CA-28111 Hormonal Regulation of Kidney Epithelial Cell Growth 
Taub University of California San Diego 



ROl-CA-28376 
Silber 



Nucleotide Biosynthesis and Degradation 
New York University 



27 



ROl-CA-28725 
Schuster 



Asparagine Biosynthesis in Normal and Tumor Cells 
University of Nebraska Lincoln 



ROl-CA-28781 Dolichyl Phosphate Biosynthesis in Tumor Cells 
Adair University of South Florida 

R23-CA-28973 A Cell Culture Model for Regulation of Tumor Cell Growth 
Epstein Children's Hospital Medical Center 

ROl-CA-29187 Characterization of a Novel Rat Lactate Dehydrogenase 
Anderson University of Pittsburgh 

ROl-CA-29307 Control of Protease Action on Human Cells 
Baker University of Kansas Lawrence 



PEPTIDE HORMONES 



ROl-CA-02146 
Bucher 



Cytoplasmic Factors in Cellular Growth 

Massachusetts General Hospital 



ROl-CA-07535 
Macleod 

ROl-CA-11685 
Orth 

ROl-CA-16417 

Ramachandran 



Control of Pituitary Gland and Pituitary Tumor Hormones 
University of Virginia 

Tumor Cell Synthesis and Secretion of Peptide Hormones 
Vanderbilt University 

Pituitary Hormones in Normal and Neoplastic Growth- 
University of California San Francisco 



ROl-CA-17309 
Levine 

ROl-CA-18406 
Baylin 

ROl-CA-19234 
Laris 

ROl-CA-22394 
Thompson 

ROl-CA-23185 
Kourides 

ROl-CA-24050 

Richardson 

ROl-CA-24604 
Surks 

ROl-CA-25614 
Williams 



Prostaglandins and Enzymes in Normal and Malignant Tissues 
Brandeis University 

Histamlnase and Amines in Neural Crest Tumors 
Johns Hopkins University 

Extrinsic Fluorescence and Membrane Potentials 

University of California Santa Barbara 

Hormonal Control of Cell Proliferation 

University of South Carolina at Columbia 

Regulation of Alpha and Beta Subunits of TSH 

Sloan Kettering Institute for Cancer Res. 

ACTH Secretion by Pituitary Tumor Cells in Culture 
Harvard University 

Triiodothyronine Receptors and Nonthyroidal Diseases 
Montefiore Hospital and Medical Center 

Antiestrogen Action and Cancer 

Worcester Fdn. for Exper. Biology 



28 



ROl-CA-25820 Receptors and Growth Factors for Neoplastic Cells 
Schlessinger Weizmann Institute of Science 



ROl-CA-28218 
Biswas 

ROl-CA-28826 
Ivarie 



ROl-CA-29808 
Iyengar 



ROl-CA-30388 
Golde 

ROl-CA-30393 
Fuller 



Hormone Production by Pituitary Tumor Cells 
Harvard University 

Mutations Affecting Gene Expression in Tumor Cells 
University of Georgia 



ROl-CA-29467 The Biochemistry of Cellular Transformation 
Vanaman Duke University 



Molecular Mechanism of Desensitization 
Baylor College of Medicine 



ROl-CA-30253 A Study of Tropic Hormone Action in Carcinoma Cells 

Mason University of Texas Hlth. Sci. Ctr. Dallas 



Humoral Regulation of Normal and Malignant Hemopoiesis 
University of California Los Angeles 

Endocrine Regulation of Melanoma Cell Differentiation 
Texas Tech University 



STEROIDS 



ROl-CA-08315 Steroid Induced Changes in Cultured Malignant Cells 
Melnykovych University of Kansas 

ROl-CA-13103 Metabolism and Inhibition of Prostatic Neoplasia 
Mawhinney West Virginia University 

ROl-CA-13410 Mechanism of Hormone Action on Target Cells in Culture 
Sonnenschein Tufts University 

ROl-CA-16091 Biochemical Control in Adrenocortical Carcinoma Cells 
Sharma University of Tenn. Center Health Scien. 

ROl-CA-17229 Keloids: An In Vitro Model of Tumor Growth Regulation 
Russell Meharry Medical College 

ROl-CA-17323 Glucocorticoid-Resistant Leukemic Lymphocytes 
Munck Dartmouth College 

ROl-CA-19907 Physiology of Pituitary Cell Glucocorticoid Binding 
Harrison Vanderbilt University 



ROl-CA-23248 
Hymer 

ROl-CA-23603 
Ascoli 



Prolactin Cell Function in Breast Cancer 

Pennsylvania State University Univ. Park 

Gonadotropin Actions in Leydig Tumor Cells 
Vanderbilt University 



29 



ROl-CA-23921 Biochemical and Clinical Aspects of Steroid Receptors 
Colas University of Wisconsin Madison 



ROl-CA-24228 
Zepp 



Endocrine Factors in Cancer- Induced Lipolysis 

Kirksville College of Osteopathic Med. 



ROl-CA-24347 Hormone Effects on Proliferation of Malignant Thymocytes 
Thompson • University of South Carolina 



ROl-CA-25365 

Gerschenson 



Hormonal Regulation of Cultured Endometrial Cells 

University of Colorado Hlth. Sciences Ctr. 



R23-CA-25655 Protein Mediators of Lethal Glucocorticoid Effects 
Nicholson University of Rochester 



RGl-CA-26020 

Braunschweig 



Corticosteroids — Cytokinetic and Biochemical Studies 
;er Allegheny-Singer Research Corporation 



ROl-CA-26112 Effect of Estrogen on Normal and Abnormal Cell Growth 
Clark Baylor College of Medicine 

ROl-CA-26617 Estrogen Mediated Pituitary Tumor Cell Growth 

Sirbasku University of Texas Hlth. Sci. Ctr. Houston 

ROl-CA-26638 Cyclic AMP:, Role in Adrenal Tumor Steroidogenesis 
Moyle Rutgers Medical School 

ROl-CA-27702 Sex Hormones, Cancer and the Immune System 

Siiteri University of California San Francisco 



ROl-CA-27828 

Eisenfeld 



Pituitary Steroid Receptors, Estrogens, and Adenomas 
Yale University 



R23-CA-28580 Therapeutic Implications of Hormone in Nevi and Melanoma 
Chaudhuri University of Illinois Medical Center 

ROl-CA-29324 Interactions of Estrogen Receptor with Chromatin 
Hilf University of Rochester 

ROl-CA-29485 Actions of Estrogens and Antiestrogens in the Nucleus 
Kallos Hospital for Joint Diseases Ortho Inst. 

ROl-CA-29497 An Adrenal Tumor: Cytochrome P-450 and Steroidogenesis 
Hall Worcester Fdn. for Exper. Biology 



ROl-CA-30380 
Young 



Glucocorticoid Suppression of Transformed Cell Growth 
Boston University 



MEMBRANOUS ORGANELLES 



ROl-CA-06576 Biological Effects of a Placental Protease Inhibitor 
Novikoff Yeshiva University 



t 



30 



ROl-CA-08964 
Racker 

ROl-CA-10951 
Pedersen 

ROl-CA-11766 
Cockrell 

ROl-CA-12312 
Clayton 

ROl-CA-12858 
Stahl 

ROl-CA-13814 
Nass 

ROl-CA-16527 

Robberson 

ROl-CA-20454 
Chan 

ROl-CA-22031 
Singer 

ROl-CA-25360 

Lehninger 

ROl-CA-27117 
Knowles 



Energy Metabolism in Normal and Tumor Cells 
Cornell University 

Control of Enzymatic Phosphate Transfer in Mitochondria 
Johns Hopkins University 

Metabolic Regulation in Neoplasms by Cation Transport 
St. Louis University 

Mitochondrial Gene Expression in Malignant Cells 
Stanford University 

Lysosome Biogenesis: Normal and Tumor Cells 
Washington University 

Mitochondrial Genetic System in Normal and Cancer Cells 
University of Pennsylvania 

Regulation of Mitochondrial DNA 

University of Texas Cancer Center 

Adenine Nucleotide Translocation in Tumor Mitochondria 
Syracuse University Syracuse 

Trans-membrane Control in Transformed and Normal Cells 
University of California San Diego 

Respiration-Coupled Transport Processes in Tumor Cells 
Johns Hopkins University 

A Study of Membrane Bound ATPases of Human Tumors 
University of California San Diego 



ROl-CA-28677 Transport in Cholesterol-Rich Tumor Mitochondria 
Coleman New York University 



RIBOSOMES 



ROl-CA-04186 Molecular Structure of Nucleic Acids and Proteins 
Rich Massachusetts Institute of Technology 



ROl-CA-08416 
Penman 



Transcription and Translation in Mammalian Cells 

Massachusetts Institute of Technology 



ROl-CA-16608 Translation Control in Reticulocytes and Leukemic Cells 
Hardesty University of Texas Austin 

ROl-CA-21663 Intermediary Metabolism in Animals and in Man 
Henshaw University of Rochester 

ROl-CA-23632 RNA Methylation: A Paradox in Normal and Cancer Cells 
Sitz Old Dominion University 



31 



ROl-CA-25618 rDNA-Binding Proteins and Control of rDNA in Tumors 
Bearden University of Hawaii at Manoa 

ROl-CA-28513 Structure of Hypomethylated Tumor 5.8S Ribosomal RNA 
Lightfoot Eastern Washington University 



M-RNA 



ROl-CA-12550 
Martin 

ROl-CA-13175 
Bottman 

ROl-CA-17287 
Stark 

ROl-CA-18065 
Edmonds 

ROl-CA-19535 
Mo Hoy 

ROl-CA-22302 
Lucas 

ROl-CA-23226 
Fausto 

ROl-CA-24165 
Zucker 

ROl-CA-24206 
Linney 

ROl-CA-24273 
Rovera 

ROl-CA-24635 
Melera 

ROl-CA-25078 
Jacob 

ROl-CA-26073 

Augenlicht 

ROl-CA-26790 
Peterson 

ROl-CA-27932 
Solter 



RNA Synthesis and Transport in Mammalian Cells 
University of Chicago 

Control of RNA Processing in Normal and Tumor Cells 
Michigan State University 

Aspects of Control in Mammalian Gene Expression 
Stanford University 

Biogenesis of Messenger RNA in Animal Cells 
University of Pittsburgh 

Structure of Nuclear RNA and Messenger RNA Formation 
University of Delaware 

Analysis of Gene Regulation by Nuclear Transplantation 
State University New York Stony Brook 

Gene Expression in Regenerating and Neoplastic Livers 
Brown University 

Hemoglobin Studies in Friend Leukemia 

Papanicolaou Cancer Research Institute 

Embryonal Carcinoma Growth and Differentiation 

La Jolla Cancer Research Foundation 

Expression of Globin Genes — Erythroleukemia Cells 

Wistar Institute of Anatomy and Biology 

Chromosomal Manifestations of Gene Expression 

Sloan Kettering Institute for Cancer Res. 

Poly(A) Polymerases from Liver and Hepatomas 

Pennsylvania State Univ. Hershey Med. Ctr. 

Mapping Globin Sequences in Nuclear Ribonucleoprotein 
Sloan Kettering Institute for Cancer Res. 

Phenotypic Variation and Neoplastic Progression 

Children's Hospital Med. Ctr. Northern Ca. 

Developmentally Regulated Genes in Teratocarcinoma 

Wistar Institute of Anatomy and Biology 



t 



32 



ROl-CA-28555 Neoplastic Activation of Fetal Gene Expression 

Hoch Scripps Clinic and Research Foundation 

ROl-CA-30124 Role of 2-5A in Growth Arrest and Hormone Responses 
Stark Stanford University 

ROl-CA-30151 Regulation of Albumin Synthesis by Amino Acids 

Ledford Medical University of South Carolina 



T-RNA 



ROl-CA-10922 Metabolism of Animal Cell Ribosomes 

Vaughan University of Pittsburgh 

ROl-CA-20683 Control Mechanisms in Human Tumor Cells — Small RNAs 
Eliceiri St. Louis University 

ROl-CA-20919 tRNA Q-Base: Its Relation to Differentiation 

Katze University of Tenn. Center Health Scien. 

ROl-CA-21245 Structure and Modification of Mammalian Transfer RNA 
Penhoet University of California Berkeley 

ROl-CA-23363 tRNA Methylation in Mammalian Cells 

Leboy University of Pennsylvania 

ROl-CA-23536 Molecular Biology of Ethionine Carcinogenesis 

Borek ■ AMC Cancer Research Center and Hospital 

ROl-CA-24727 Changes in RNA Metabolism and the Induction of Tumors 
Apirion Washington University 

ROl-CA-26423 Modification of tRNA 1/4YS Controls Cell Proliferation 
Ortwerth University of Missouri Columbia 

ROl-CA-27235 tRNA Modification and Gene Expression in Mammalian Cells 
Hatfield University of California Irvine 

ROl-CA-27288 Regulation of Gene Expression in Hximan Tumor Cells 
Eliceiri St. Louis University 

ROl-CA-28053 Thiolated tRNAs in Rat Liver and Morris Hepatomas 
Wong University of Chicago 

ROl-CA-28395 tRNA Methylation in Normal and Neoplastic Rat Tissues 
Leboy University of Pennsylvania 



33 



ROl-CA-14835 
Korn 

ROl-CA-15044 

Manuelidis 

ROl-CA-15187 
Baril 



DNA 



DNA Polymerases in Normal and Neoplastic Human Cells 
Stanford University 

Pathogenetic Determinants of Human CNS Tumors 
Yale University 

DNA Synthesis: Regulation in Normal and Cancer Cells 
Worcester Fdn. for Exper. Biology 



ROl-CA-16790 DNA Transcription Control in Normal and Cancer Cells 
Maio Yeshiva University 

ROl-CA-17723 Regulation of DNA Synthesis in the Novikoff Hepatoma 
Meyer University of Cincinnati 

ROl-CA-18138 Mammalian Polyamine Metabolism 

Pegg Pennsylvania State Univ. Hershey Med. Ctr. 

ROl-CA-18564 Chediak-Higashi and Cancer Cells with Similar Defects 
Oliver University of Connecticut Health Center 



ROl-CA-22610 
Brinkley 

ROl-CA-23365 
Chang 

ROl-CA-24151 

Vandenbark 

ROl-CA-24158 
Collins 

ROl-CA-24323 
Mathews 

ROl-CA-24845 
Loeb 

ROl-CA-25023 
Gottlieb 



Cytoskeleton and Cell Transformation to Malignancy 
Baylor College of Medicine 

Function of DNA Poljnnerases in Normal and Cancer Cells 

U.S. Uniformed Services Univ. of Health Sciences 

Regulation of Cell Growth by 5 '-Methylthioadenosine 

University of Oregon Hlth. Sciences Ctr. 

DNA Synthesis in Transformed Cells 

Virginia Commonwealth University 

DNA Precursor Dynamics in Animals Cells 
Oregon State University 

The Fidelity of DNA Replication in Human Lymphocytes 
University of Washington 

Inhibitor of DNA Polymerase Produced by Tumor Cells 
Tulane University of Louisiana 



ROl-CA-26391 Molecular Pathology of Leukemia and Lymphoma 
Coleman University of Kentucky 



GROWTH FACTORS 



ROl-CA-11176 Factors Required for Mammalian Cell Division 

Holley Salk Institute for Biological Studies 



i 



34 



ROl-CA-13808 
Wood 



ROl-CA-17620 
Smith 



Transformation and Recovery of Crown Gall Tumor Cells 
Rockefeller University 



ROl-CA-14019 Tumor Angiogenesis : A Control Point in Tumor Growth 
Folkman Children's Hospital Medical Center 

ROl-CA-17203 Role of Serum and Cations in Malignant Cell Growth 
Tupper Syracuse University 



Growth Control in Normal and Neoplastic Cells 
University of Nebraska Lincoln 



ROl-CA-19275 
Dodge 



Normal and Leukemic Granulopoiesis 
Wake Forest University 



ROl-CA-19731 Hormonal Requirement of Cells In Vitro and In Vivo 
Sato University of California San Diego 

ROl-CA-21763 Growth Control by a Mitogen Purified from Cartilage 
Klagsbrun Children's Hospital Medical Center 



ROl-CA-22410 
Linder 



Copper Absorption and Ceruloplasmin in Cancer 

California State University Fullerton 



ROl-CA-23528 

Broxmeyer 



Inhibitory Interactions Regulating Hematopoiesis 

Sloan Kettering Institute for Cancer Res. 



ROl-CA-24395 Fibroblast Growth Factors and Phosphorylation 

Nilson-Hamilton Salk Institute for Biological Studies 



ROl-CA-27031 
Lim 



Maturation Factors and Cancer Cells 
University of Chicago 



ROl-CA-27113 Molecular Analysis of Platelet-Derived Growth Factor 
Scher Sidney Farber Cancer Institute 

ROl-CA-27217 Growth Factors and Receptors in Chemical Transformation 
Moses Mayo Foundation 

R23-CA-27802 Pharmacologic Modulation of Erythroid Colony Formation 
Beckman Tulane University of Louisiana 

ROl-CA-28110 Nerve Growth Factor: Secretion by Cancer Cells 
Young University of Florida 



ROl-CA-28456 
Rifkin 



Biochemical Mechanisms of Cellular Invasion 
New York University 



ROl-CA-28540 Growth and Migration of Capillary Endothelial Cells 
Zetter Children's Hospital Medical Center 

ROl-CA-28638 Altered Nutritional Requirements for Growth 
Topp Cold Spring Harbor Laboratory 



35 



ROl-CA-28858 
Pickart 

ROl-CA-29101 

LaBrecque 

ROl-CA-30101 

Antoniades 



Biological and Synthetic Modulators of Cell Growth 
Virginia Mason Research Center 

Characterization of a Liver Specific Growth Promoter 
University of Iowa 

Structure and Function of Platelet-Derived Growth Factor 
Center for Blood Research 



ROl-CA-30479 Mononuclear Phagocyte-Derived Growth Regulating Factors 
Gillespie University of North Carolina Chapel Hill 

ROl-CA-30536 New Myeloid Hemapoietins : Normal and Leukemic Marrow 
Wells University of California Los Angeles 



NUCLEUS 



ROl-CA-10872 
Isenberg 

ROl-CA-12226 
Paik 

ROl-CA- 12877 
Langan 

ROl-CA-13195 
Smulson 

ROl-CA-15135 
Zweidler 



Interactions of Chromosomal Proteins 
Oregon State University 

Protein Methylation in Neoplastic Tissue 
Temple University 

Function of Lysine Rich (HI) Histone Phosphorylation 

University of Colorado Hlth. Sciences Ctr. 

Histone ADP-Ribosylation and HeLa Cell Replication 
Georgetown University 

Histones in Cell Differentiation and Carcinogenesis 
Fox Chase Cancer Center 



ROl-CA-15923 
Shelton 

ROl-CA-16910 
Rowley 

ROl-CA-17782 
Reeck 

ROl-CA-18389 
Hnilica 

ROl-CA-18455 

Wray 

ROl-CA-21927 
Maizel 

ROl-CA-24546 
Kornberg 



Nonhistone Proteins of Reactivated Erythrocyte Nuclei 
Virginia Commonwealth University 

Chromosome Aberrations in Myeloproliferative Diseases 
University of Chicago 

Tumor- Enriched Nonhistone Chromatin Proteins 
Kansas State University 

Proteins of the Cell Nucleus 

Vanderbilt University 

Isolated Chromosomes in Genetics and Cancer Research 
Baylor College of Medicine 

Chromatin Structure of Normal and Malignant T-Cells 
University of Texas Cancer Center 

Relation of Histones to DNA in Normal and Cancer Cells 
Stanford University 



36 



ROl-CA-25055 
Hecht 



Cytogenetics of Clonal Neoplasias 

Southwest Biomedical Research Institute 



ROl-CA-25735 Regulatory Processes Biochemistry in Synchronized Cells 
Hodge Medical College of Georgia 

ROl-CA-27446 Nuclear Proteins in Hepatoma and Normal Liver 

Ruoslahti La Jolla Cancer Research Foundation 



ROl-CA-27661 
Palmer 



Sister Chromatid Exchange in ALL 

Indiana Univ. -Purdue Univ. at Indianapolis 



ROl-CA-28168 Cytogenetics of Hepatocellular Carcinoma in Nigeria 

Cervenka University of Minnesota of Minneapolis-St . Paul 

ROl-CA-28679 Chromosomal Organization of Dihydrof olate Reductase Gene 
Biedler Sloan Kettering Institute for Cancer Res. 

ROl-CA-29340 rDNA Distribution in Chromosomes of Neoplastic Cells 
Henderson Columbia University 

ROl-CA-29476 Clonal Karyotypic Evolution in Human Solid Tumors 
Trent University of Arizona 

ROl-CA-29617 Role of Double Minutes and HSR Markers in Tumor Cells 
George Johns Hopkins University 

ROl-CA-29779 DNA Replication: Chromosomes and Neoplasms 

Cervenka University of Minnesota of Minneapolis-St. Paul 

ROl-CA-31024 Fine Structural Chromosomal Defects in Acute Leukemia 

Yunis University of Minnesota of Minneapolis-St. Paul 



CONTRACTILE ELEMENTS 



ROl-CA-05493 Leukopoietic Mechanisms 

De Bruyn University of Chicago 

ROl-CA-15229 Actin and Tubulin Cancer Cells 

Weihing Worcester Fdn. for Exper. Biology 

ROl-CA-15544 Effect of Microtubular Proteins on Cell Surfaces 

Berlin University of Connecticut Health Center 



RO1-CA-20836 
Godman 

ROl-CA-26473 
Mattson 



Cytochalasins and Cell Membrane Contractile Apparatus 
Columbia University 

ACTH Effects of the Cytoskeleton of Adrenal Tumor Cells 
Case Western Reserve University 



ROl-CA-26867 
Zeldman 



Invasion and Metastasis 

University of Pennsylvania 



37 



ROl-CA-27458 
. Tuszynski 

ROl-CA-28362 
Rapp 

ROl-CA-29985 

Weisenberg 



Biochemistry of Cytoskeletal Proteins 
Temple University 

Levels of Metastasis Inhibition in Primary Cancers 
Roswell Park Memorial Institute 

Microtubules and Nonmicrotubular Aggregates 
Temple University 



ROl-CA-30085 Immune Mechanism in Cells Initiating ERV Production 
Albrecht-Buehle Cold Spring Harbor Laboratory 



DEVELOPMENT AND DIFFERENTIATION 



ROl-CA-02662 Investigations of Teratocarcinogenesis 
Stevens Jackson Laboratory 

ROl-CA-08482 Differentiation and Function of Hematopoietic Cells 
Scott Virginia Commonwealth University 

ROl-CA-10095 Gene Action and Cellular Differentiation in Culture 
Silagi Cornell University Medical Center 

ROl-CA-12187 Growth and Differentiated Function in Mammalian Cells 
Green Massachusetts Institute of Technology 

R01-CA-13047 Control Mechanisms of Differentiation and Malignancy 
Friend Mount Sinai School of Medicine 



ROl-CA-13 533 
Sussman 

ROl-CA-14319 

Schengrund 

ROl-CA-15222 

Smith 

ROl-CA-16368 

Skoultchi 

ROl-CA-16746 
Taylor 

ROl-CA-17389 
Wolfe 

ROl-CA-17563 
Hoch 

ROl-CA-17575 
Housman 



Ectopic Placental Proteins in Human Cancer 
Stanford University 

Biological Functions of Plasma Membrane Sialidase 

Pennsylvania State Univ. Hershey Med. Ctr. 

Hepatoma Alpha-Fetoprotein: Chemistry and Metabolism 

University of Vermont and St. Agric. College 

Control of Differentiation of Erythroleukemic Cells 
Yeshiva University 

Control of Albumin Synthesis in Liver and Hepatomas 

Pennsylvania State Univ. Hershey Med. Ctr. 

C-Cell Hyperplasia and Thyroid Medullary Carcinoma 
Tufts University 

DNA Binding Proteins in Human Serum 

Scripps Clinic and Research Fdn. 

Erythroid Differentiation in Friend Leukemia Cells 
Massachusetts Institute of Technology 



i 



38 



ROl-CA-18375 

Goldwasser 

ROl-CA-19492 . 
Coleman 



ROl-CA-20053 
Me ins 

ROl-CA-21566 
Kuettner 

ROl-CA-22099 
Kelleher 

ROl-CA-22227 
- Sell 

ROl-CA-22294 
Kinkade 

ROl-CA-22556 
Burgess 

ROl-CA-22558 
Oegema 

ROl-CA-22670 
Trentin 



Hemopoietic Stem Cells and Induced Differentiation 
University of Chicago 

Terminal Transferase in Mammalian Hemopoietic Tissue 
University of Kentucky 

Tumor Inception and Progression 

University of Illinois Urbana-Champaign 

Antittimor Invasion Factors Derived from Cartilage 

Rush-Presbyterian-St . Luke's Medical Ctr. 

Alpha-Fetoprotein Synthesis by Hepatoma Cells In Vitro 

University of Vermont and St. Agric. College 

Onco-Developmental Gene Control: Alpha-Fetoprotein 
University of California San Diego 

Quantitative Studies on Granulocyte Differentiation 
Emory University 

Differentiation of Granulocytes and Monocytes 

Walter and Eliza Hall Inst. Medical Res. 

Comparative Biochemistry of Cartilage Tumors 

University of Minnesota of Minneapolis-St . Paul 

Control of Hemopoietic Stem Cell Differentiation 
Baylor College of Medicine 



ROl-CA-22735 Thymic Modification of Leukemogenesis 
Sensenbrenner Johns Hopkins University 

ROl-CA-23097 Embryo Derived Teratocarcinoma 

Damjanov Hahnemann Med. Col. and Hosp. of Philadelphia 

ROl-CA-23615 Molecular Basis of Differentiation and Neoplasia 
Roeder Washington University 

ROl-CA-24010 Differentiation of Cancer 

Pierce University of Colorado Hlth. Sciences Ctr. 



ROl-CA-24241 

Sandquist 

ROl-CA-24479 
Chen 

ROl-CA-25098 
Chiu 

ROl-CA-25164 

Sudilovsky 



Differentiation in a Malignant Neural Tumor 
University of Iowa 

Membrane Structure and Enzyme Induction 

Rutgers The State Univ. New Brunswick 

Alpha-Fetoprotein Regulation in Fetal and Cancer Liver 

University of Vermont and St. Agric. College 

Malignant Phenotypes: Cytoplasmic and Nuclear Control 
Case Western Reserve University 



39 



R23-CA-25285 Ectopic Antigens in Hepatocellular Cancer 

Higgins Sloan Kettering Institute for Cancer Res. 



ROl-CA-25512 
Brennan 



Modulators of Granulopoiesis from Human Cell Lines 
University of Rochester 



ROl-CA-25799 
Anderson 



Conformation of Alpha-Fetoprotein Gene in Chromatin 
Purdue University 



ROl-CA-25966 X-Chromosome Activity in Teratocarcinoma Stem Cells 
Martin University of California San Francisco 



ROl-CA-25972 
Metcalf 



Leukemia Induction in Multipotential In Vitro Colonies 
Walter and Eliza Hall Inst. Medical Res. 



ROl-CA-25985 

Christman 



Response to Phagocytic Leukocytes to Tumor Promoters 
Mount Sinai School of Medicine 



ROl-CA-26014 

Rosenstraus 



Cell Interactions in Teratocarcinoma Differentiation 
Rutgers The State Univ. New Brunswick 



ROl-CA-26038 Study of Myeloid Leukemia Using Human Leukemia Lines 
Koeffler University of California Los Angeles 



ROl-CA-26105 

Sytkowski 



Human Renal Cancer and Hematopoiesis 

Children's Hospital Medical Center 



ROl-CA-26234 Fibroblastic Colonies in Myeloproliferative Disorders 
Greenberg University of California Davis 



R23-CA-26401 
Fay 



Human Leukemia Marrow 

Duke University 



ROl-CA-26506 Myeloid Development in an Induced Leukemic Cell Line 
Cohen Children's Hospital Medical Center 

ROl-CA-26656 Cell Culture Analysis of Human Epidermal Neoplasia 
Rheinwald Sidney Farber Cancer Institute 

ROl-CA-26847 Regulation of mRNA Translation in Animal Cells 
Weber University of South Florida 



ROl-CA-26993 
Sheid 



Cytotoxic Factor(s) from Human Ovarian Adenocarcinoma 
Downstate Medical Center 



ROl-CA-27287 Vaginal and Cervical Epithelial Cell Culture Model 
Rice Harvard University 



ROl-CA-27466 

Quesenberry 



Endothelial Colony-Stimulating Activity 
University of Virginia 



ROl-CA-27580 
Oshima 



Chromatin Proteins of Embryonal Carcinoma Cells 

Massachusetts Institute of Technology 



40 



ROl-CA-27682 New Technology for Classifying Human Leukemia 

Stang University of California Los Angeles 

ROl-CA-28050 Regulation of Alpha-Fetoprotein Gene Expression 
Tilghman Institute for Cancer Research 

ROl-CA-28106 Differentiation of Teratocarcinoma Cells 

Levine State University of New York Stony Brook 

ROl-CA-28179 Lithium Effects on Hemopoietic Stem Cells 
Joyce University of Pittsburgh 

ROl-CA-28228 Expansion of Hemopoietic Bone Marrow 
Lee University of Washington 

R23-CA-28289 Differentiation and Tumorigenicity in Recombinant Cells 
Moore University of Colorado Hlth. Sciences Ctr. 

ROl-CA-28427 EGF and Its Receptors in Embryonic Differentiation 
Adamson La Jolla Cancer Research Foundation 

R23-CA-28512 Regulation of Normal and Leukemic Myeloid Stem Cells 

Pelus Sloan Kettering Institute for Cancer Res. 

ROl-CA-28656 Differentiation of Capillary Endothelial Cells 
Auerbach University of Wisconsin Madison 

ROl-CA-28815 Biosynthesis of Carcinoembryonic Antigen 
Mendicino University of Georgia 

ROl-CA-29169 Gene Expression Embryonal Carcinoma Differentiation 
Linney La Jolla Clinic Research Foundation 

ROl-CA-29895 Antiproliferative Effects of Interferons 

Baglioni State University of New York at Albany 

ROl-CA-29959 Glycoproteins in Differentiation and Oncogenesis 

Fukuda Fred Hutchinson Cancer Research Center 

ROl-CA-30049 Oncofetal Gene Regulation in Hepatocarcinogenesis 

Papaconstantino University of Texas Med. Br. Galveston 

ROl-CA-30684 Regional Differences in Tumor Growth and Development 
Auerbach University of Wisconsin Madison 

ROl-CA-31042 Developmental Regulation of E Globin Gene Expression 
Lo University of Pennsylvania 

ROl-CA-31271 Differentiation and Stroma- Induction in Neural Tumors 
Rubinstein University of Virginia 

ROl-CA-31777 Budr Dependence, Malignancy, and Differentiation 
Davidson Children's Hospital Medical Center 



41 



CELL GROWTH 



ROl-CA-06663 Repair of Chromatin by 3-Methyladenine N-Glycosylase 
Lieberman University of Pittsburgh 

ROl-CA-08373 Study of Factors Controlling Cellular Proliferation 
Baserga Temple University 

ROl-CA-15062 Studies of Normal and Neoplastic Prostate 

Ahmed University of Minnesota of Minneapolis-St . Paul 

ROl-CA-15141 Control of Nuclear Events in Normal and Neoplastic Cells 
O'Neill University of Utah 

ROl-CA-15305 Effect of Malignancy on Cell Growth Requirements 
Ham University of Colorado at Boulder 

ROl-CA-15381 Control of Tumor Induced Angiogenesis 
Fenselau Johns Hopkins University 

ROl-CA-15744 Primary Events in Regulating Cell Multiplication 
Rubin University of California Berkeley 

ROl-CA-15813 Lipid Transport and Metabolism in Cancer-Host Systems 
Baker University of California Los Angeles 

ROl-CA-16463 Triiodothyronine Receptors and Thyrotroph Neoplasia 
Surks Montefiore Hospital and Medical Center 

ROl-CA-16816 Mechanism of Chemical Carcinogenesis In Vitro 
Moses Mayo Foundation 

ROl-CA-19126 Cystine Auxotrophy in Human Malignancy — Implications 
Lazarus Sidney Farber Cancer Institute 

ROl-CA-20136 Tumor Lipids: Metabolism and Structural Studies 

Wood Texas Agri. and Mech. Univ. College Station 



ROl-CA-21359 
Bertram 



Cell Interactions During Malignant Transformation 
Roswell Park Memorial Institute 



ROl-CA-22011 Studies of Myeloid Leukemia in RFM Mice 

Trobaugh Bush-Presbyterian-St . Luke's Medical Ctr. 

ROl-CA-22042 Role of Serxom in the Foreign Body Response 
Stiles Sidney Farber Cancer Institute 

ROl-CA-22088 Control of Growth in Quiescent Human Cells 

Norin Montefiore Hospital and Medical Center 

ROl-CA-23022 Studies of Mitosis in Normal and Neoplastic Cells 
Brinkley Baylor College of Medicine 



42 



ROl-CA-24193 Regulation of Manmalian Cell Cycle 

Pledger University of North Carolina Chapel Hill 

ROl-CA-24385 Effects of Phorbol Esters on Lymphocyte Stimulation 

Mastro Pennsylvania State University Univ. Park 

ROl-CA-24914 Control of Cell Division in Leukemia Cells 

Wolcott Louisiana State Univ. Med. Ctr. Shreveport 

ROl-CA-24961 Protein Synthesis and Control of the Cell Cycle 
Castor Institute for Cancer Research 

ROl-CA-25009 Role of Disulfides in Normal and Tumor Cell Growth 
Fahey University of California San Diego 

ROl-CA-26070 Control of Cycle Progression in Animal Cells 
Basilico New York University 

ROl-CA-26081 Cell Cycle Dependence of Cell Surface Receptors 
Varga Yale University 

R23-CA-26889 Regulation of Cell Growth by Membrane Ion Flux 
Frantz Sidney Farber Cancer Institute 

R23-CA-26956 Detailed Cell Kinetic Analyses of Human Neuroblastoma 
Neely Children's Hospital Medical Center 

ROl-CA-27151 Regulation of Cell Replication by Thymidine 

Reiter University of Illinois Medical Center 

ROl-CA-27399 Regulation of Mitosis in Normal and Transformed Cells 
Sisken University of Kentucky 

ROl-CA-27419 Immunopharmacologic Consequences of Tumor Heterogeneity 
Heppner Michigan Cancer Foundation 

ROl-CA-27544 Purification and Characterization of Mitotic Factors 
Rao University of Texas System Cancer Center 

ROl-CA-27562 Regulation of Transfer RNA Levels in Mammalian Cells 
Litt University of Oregon Hlth. Sciences Ctr. 

ROl-CA-27564 Methionine Dependence — A Metabolic Marker in Cancer 
Hoffman University of California San Diego 

R23-CA-27808 Plasticity of Chromaffin and Pheochromocytoma Cells 
Tischler Tufts University 

ROl-CA-27809 Pathways of Energy Metabolism in Malignancy In Vivo 
Sauer Mary Imogene Bassett Hospital 

ROl-CA-28140 A Novel Vitamin B6 Metabolite in Hepatomas 
Tryfiates West Virginia University 



43 



ROl-CA-28238 Effects of Mitogens on Normal and Neoplastic Cells 
Vogel University of Washington 

ROl-CA-28240 Pathology in Cell Cycle Control of Differentiation 
Scott Mayo Foundation 

R23-CA-28329 Phase Specific DNA Repair in Irradiated Tumor Cells 
Keng University of Rochester 

ROl-CA-28519 Characterization of Cells and Clones from Human Brain 
Rosenblum University of California San Francisco 

ROl-CA-28760 Anticoagulants, Vitamin K and Tumor Cell Growth 
Hauschka Children's Hospital Medical Center 

ROl-CA-28803 The Role of AP4A in Malignant Transformation 
Rapaport Boston University 

ROl-CA-28964 Tumor Cell-Vascular Endothelial Cell Interactions 
Birdwell La Jolla Cancer Research Foundation 



ROl-CA-29560 
Wicks 

ROl-CA-30083 
Eisenst 

ROl-CA-31053 

Vogelstein 



Cyclic AMP Analogs as Growth Regulators in Tumor Cells 
University of Tennessee Knoxville 

Aortic Growth Inhibitors 

Mount Sinai Medical Center 

Mitotic Inducing Protein (S) from Mammalian Cells 
Johns Hopkins University 



SOMATIC CELL GENETICS 



ROl-CA-12130 
Harris 

ROl-CA-16631 
Meiss 

ROl-CA-16720 
Klinger 

ROl-CA-16754 

Littlefield 

ROl-CA-19401 

Stanbridge 

ROl-CA-21054 
Shin 

ROl-CA-21365 
Sager 



Cytoplasmic Inheritance in Normal and Tumor Cells 
University of California Berkeley 

Isolation and Analysis of DNA Mutants of BHK Cells 
New York University 

Gene Regulation and Interaction — Normal and Malignant Cells 
Yeshiva University 

Hybridization, DNA Function, Mutation in Cell Culture 
Johns Hopkins University 

Genetic Analysis of Human Malignancy 

University of California Irvine 

Genetic Analysis of Malignant Transformation 
Yeshiva University 

Cytoplasmic Genes in Normal and Tumorigenic Cells 
Sidney Farber Cancer Institute 



44 



ROl-CA-23003 Regulation of Expression of Viral Transformation 
Ozer Hunter College 



ROl-CA-24828 
Sager 

ROl-CA-25342 

Siniscalco 

ROl-CA-27607 
Lee 



Genetic Analysis of Tumorigenesis 

Sidney Farber Cancer Institute 

Complement of Sister Chromatid Exchange to Cell Hybrids 
Sloan Kettering Institute for Cancer Res. 

Coordinated Control of Mammalian Gene Expression 
University of Southern California 



ROl-CA-27712 Gene Expression During Mammalian Development 

Croce Wistar Institute of Anatomy and Biology 

ROl-CA-27713 Gene Expression During Mammalian Development 
Illmensee University of Geneva 

ROl-CA-28559 Study of Malignant Transformation: A Genetic Analysis 
Athwal College of Medicine and Dentistry of NJ 

ROl-CA-30938 Structural and Functional Analysis of Cloned MHC Gene 
Weissman Yale University 



INHERITANCE OF NEOPLASMS 



R23-CA-28963 Genetic Basis of SJL/J Murine Reticulum Cell Sarcoma 
Bubbers University of California Los Angeles 

ROl-CA-29078 Cellular Origins of Rat Hepatic Preneoplasias 
lannaccone Northwestern University 

R23-CA-29944 Analysis of Genetic Heterogeneity in Fanconi Anemia 

Auerbach Sloan Kettering Institute for Cancer Res. 



PLASMIDS, VIRUSES 



ROl-CA-11526 Tximor- Inducing Substance of Agrobacterium Tumefaciens 
Kado University of California Davis 

ROl-CA-13015 Molecular Basis of Crown Gall Tumorigenesis 
Nester University of Washington 

ROl-CA-13015 Molecular Basis of Crown Gall Tumorigenesis 
Nester University of Washington 

RO1-CA-14026 Tumor Virus, Plasmid and Cell Analysis 

Goodman University of California San Francisco 



45 



ROl-CA-18604 The Mechanism of Tumorigenesis by A. Tumefaciens 

Matthysse University of North Carolina Chapel Hill 

ROl-CA-19402 Molecular Genetics of Agrobacterium Plasmids 
Farrand Loyola University Medical Center 

ROl-CA-26963 Molecular Regulation of Crown-Gall Tumor Growth 
Chang " University of Wisconsin Parkside 

R23-CA-27421 Development of a Eukaryotic Cloning Vehicle 
Rogers Oregon State University 

ROl-CA-27424 Agrobacterium DNA Insertion and Expression in Plants 
Goodman University of California San Francisco 

ROl-CA-28946 Transfection by Endogenous Human Transforming Genes 
Cooper Sidney Farber Cancer Institute 

ROl-CA-29474 Cytology, Biochemistry of Viral-Specific Proteins 
Buchanan Massachusetts Institute of Technology 



ROl-CA-29477 

Kucherlapati 



Analysis of Malignancy by Gene Transfer 
Princeton University 



IN VIVO AND IN VITRO TUMOR LINES 



ROl-CA-11683 Coenzymes and Nucleic Acids Metabolism 

Kaplan University of California San Diego 



ROl-CA-24145 
Beamer 



Ovarian Tumors in Young Mice 

Jackson Laboratory 



R23-CA-24393 Derivation of Nerve Cell Lines from the Brain 

Giotta Salk Institute for Biological Studies 

ROl-CA-24621 Induction-Continuation-Genetics of Experimental Tumor 
Morris Howard University 

RO1-CA-25630 Studies of Malignant Progression Using Human Cells 
Smith University of California Berkeley 



ROl-CA-25718 

Berkelhammer 



Swine Melanoma: In Vitro Growth and In Vivo Pathology 
Cancer Research Center 



ROl-CA-26063 Primary Culture of Prostatic Epithelial Cells 
Douglas Tufts University 

ROl-CA-26110 Androgen-Responsive Prostate Epithelial Cells 

McKeehan W. Alton Jones Cell Science Center 

ROl-CA-28641 Tumor Imaging Using ^^Tc and ^In-Labeled Leukocytes 
Dettman Brown University 



46 



ROl-CA-28668 Biologic Markers for Melanoma 

Gehrke University of Missouri Columbia 

ROl-CA-29440 Biochemical Identification of Organ Specific Melanoma 
Mehard University of California San Francisco 

ROl-CA-29550 Tumor Cell with Varying Degrees of Malignancy 
Varani University of Michigan 

ROl-CA-30082 Genetics and Development of Teratocarcinoma Cells 
Nesbitt University of California San Diego 



ROl-CA-30621 
Epstein 



Biological and Immunobiochemical Studies of Human Hemato 
Northwestern University 



CONFERENCES 



R13-CA-15961 Seminars and Workshops in Techniques of Cancer Research 
King University of Colorado Hlth. Sciences Ctr. 



R13-CA-28431 

Rosenberg 

R13-CA-30001 
Watson 

R13-CA-30222 
Ramwell 



Yale University 

Cold Spring Harbor Lab Meetings on Cell Proliferation 
Cold Spring Harbor Laboratory 

International Conference on Prostaglandins and Cancer 



R13-CA-30245 1982 Annual Symposium on Fundamental Cancer Research 
Lemaistre University of Texas System Cancer Center 

R13-CA-31383 Conference on Gene Amplification 

Watson Cold Spring Harbor Laboratory 



PROGRAM PROJECTS 



POl-CA-10893 
Busch 



Cancer Research Center 

Baylor College of Medicine 



POl-CA-12174 
Green 



Study of Growth and Cell Control Processes 

Massachusetts Institute of Technology 



POl-CA-12923 
Baserga 

POl-CA-14294 

Isselbacher 



A Correlated Study on the Biology of Neoplasia 
Temple University 

Tumor Associated Changes of Intestinal and Liver Cells 
Massachusetts General Hospital 



47 



POl-CA-14454 Membranes in Normal and Cancer Cells 
Racker Cornell University 

POl-CA-15823 Program in Developmental Biology of Cancer 

Pierce University of Colorado Hlth. Sciences Ctr. 

POl-CA-19265 Cancer Biology Research Center 

Ultmann University of Chicago 

POl-CA-20810 Somatic Cell Genetics in Cancer 

Puck University of Colorado at Boulder 

POl-CA-21901 Studies of Normal and Malignant Cell Membranes 
Roseman Johns Hopkins University 

POl-CA-22376 Control of Gene Expression: Normal and Neoplastic 
Feigelson Columbia University 

POl-CA-23052 Program Project on Athymic Mice and Human Tumors 
Kaplan University of California San Diego 

POl-CA-23076 Regulatory Mechanisms in Tumor Biology 

Mueller University of Wisconsin Madison 

POl-CA-25845 Pathobiology of Small Cell Carcinoma of the Lung 
Sorenson Dartmouth College 

POl-CA-25875 Cell Differentiation and Cancer 

Croce Wistar Institute of Anatomy and Biology 

POl-CA-26712 Glycoproteins, the Cytoskeleton and Cancer 

Hynes Massachusetts Institute of Technology 

P01-CA29545 Interferon, Differentiation and Oncogenesis 

Carter Hahnemann Med. Col. and Hosp. of Philadelphia 

POl-CA-29569 Gene Organization and Expression in Eukaryotes 
Watson Cold Spring Harbor Laboratory 



DIFFICULT TO' CATEGORIZE 



ROl-CA-09247 Partial Subsidy for the Journal of Cancer Research 

Philips American Association for Cancer Research 

ROl-CA-21607 Secretory Processes in the Prostate Gland 

Smith University of Massachusetts Medical School 

ROl-CA-25298 Biology of Human Cutaneous Malignant Melanoma 
Clark University of Pennsylvania 

R01-CA-27120 Interferon System: Action, Induction, and Regulation 
Ts'O Johns Hopkins University 



48 



ROl-CA-28571 Recognition of Patterns in Cancer-Related Sequences 
Erickson Rockefeller University 

ROl-CA-29551 Complement Mediated Tumor Cell Chemotaxis 
Ward University of Michigan 



49 



f 



50 



CONTRACT RESEARCH SUMMARY 

Title: Morris Hepatoma Resource Program 

Principal Investigator: Dr. Wayne E. Criss 

Performing Organization: Howard University College of Medicine 

City and State: Washington, DC 

Contract Number: NCI-CB-14345-39 

Starting Date: 7/1/81 Expiration Date: 6/30/84 

Goal: To maintain eleven Morris hepatomas representative of the spectrum of 
rapidly-to very slow-growing-tumors in stock rats and provide them on re- 
quest to laboratories for research purposes. 

Approach: The hepatomas will be propagated by serial transplantation in 
rats and periodically monitored by enzyme profiles and assay of specific 
metabolites to assure stability of each line. Requests for any of the hepa- 
tomas will be filled depending on availability by injecting tumor tissue into 
host rats purchased by the requestor and then shipping them to his/her labora- 
tory by air freight. 

Progress: New contract 



Significance to Cancer Research: Each of these hepatomas has specific 

characteristics that make it the tumor of choice for certain research 

projects. Currently, NCI grants in the areas of enzymology, intermediary 

metabolism and molecular biology depend upon this liver tumor system. 

Project Officer: Dr. Colette Freeman 
Program: Tumor Biology Section 
FY 81 Funds: $120,000 



51 



( 



52 



IMMUNOLOGY PROGRAM 

The role of the Immunology Program of the National Cancer Institute is 

to support studies which contribute to an understanding of the role of the 

immune system on the development, growth and spread of tumors. The specific 
areas of investigation supported by the Program include: 

1) The synthesis and structure of myeloma proteins in animals and man. 

2) The synthesis, structure, and mechanism of action of antibodies cap- 
able of reacting with tumor cells and agents which induce tumors. 

3) The synthesis, structure and mechanism of action of humoral factors 
other than antibody which participate in, activate and/or regulate 
the immune response to tumors. Factors of interest include: comple- 
ment, interferon, lymphokines, lymphoid cell growth factors, helper 
factors, suppressor factors as they are involved in immune responses 
to tumors. 

4) The immunobiology of lymphocytes that participate in anti-tumor re- 
sponses including their development, heterogeneity, interactions 
and mechanisms of action. 

5) The immunobiology of malignancies of the immune system (lymphomas and 
leukemias) including studies of immunologic markers for the classifi- 
cation and characterization of neoplastic cells and their normal 
counterparts. 

6) The immunobiology of monocytes and macrophages that participate in 
antitumor responses including their development, heterogeneity, 
interactions, and mechanisms of action. 

7) The identification, isolation and characterization of cell surface 
determinants which serve as target antigens for the immune response 
to tumors (e.g., tumor specific antigens, tumor associated antigens). 

8) The identification, isolation and characterization of cell surface 
determinants of Ijrmphocytes and macrophages which are involved in 
the responses of these cells to tumors. 

9) Immune surveillance against the development of tumors of various 
origins by all immune mechanisms (e.g., T cell immunity, macrophage 
reactivity, natural killer cell activity). 

10) Imraunopathology studies on the host-tumor interaction. 

11) Immune status of tumor bearing animals and man. 

12) Immunogenetic studies relevant to the anti-tumor immune response. 

13) Immunobiology of sarcomas, carcinomas, and melanomas. 



53 



f 

At one time, the Immunology Program was unique in touching upon all 
four of the major research thrusts of the Institute — cause and prevention, 
detection and diagnosis, treatment and lastly biology. In line with organi- 
zational changes within the Institute, the emphasis for the Immunology Program 
is now centered upon mechanistic studies with responsibility for the more 
applied immunological efforts in detection and diagnosis residing in the 
Diagnosis Program and responsibility for explicit treatment studies involving 
immunologic manipulation residing in the programs of the Division of Cancer 
Treatment. Reference to the accomplishments in these later areas should be 
sought in the reports of those programs. 

A small number of selected areas are presented to highlight the ongoing 
research efforts supported by the Immunology Program. The goal is to convey 
a general sense of the movement in the field. Recognition of all the scien- 
tists contributing to progress in Tumor Immunology within the past year would 
be too extensive to identify in this report. The specific citations used 
are limited examples of this progress. 

A. Molecular Genetics and Biology of Immunoglobulins : 

Studies on antigen-binding macromolecules continue to provide important 
insights into the biology of the immune system which are pertinent to our 
knowledge of how tumor associated antigens are recognized and how responses 
to those antigens develop and are regulated. 

Three major theories have been proposed to account for the ability of 
the immune system to react against the tremendous diversity of possible 
antigens to which we may be exposed. Recent evidence indicates that all 
three theories may be, at least in part, correct. 

An immunoglobulin molecule consists of constant (c) and variable (v) 
domains in both its light and heavy polypeptide chains. Studies using 
oligonucleotide probes and the technique of measuring duplex formation due 
to homology in base sequences have revealed the existence of only one copy 
of each gene specifying one of the several constant regions per haploid 
genome. Similar approaches toward mapping the genes which determine the 
variable domains indicate the existence of large sets of genes existing in 
closely related clusters (1). Thus, the first mechanisms to account for the 
existing diversity is the large number of variable region genes which can be 
combined with any one constant region gene in the synthesis of a specific 
antibody. 

A second mechanism for generating diversity is the rearrangement of genes 
which occurs at the DNA level as germline cells mature into somatic cells 
Such as the plasmacyte. 

Light chains of the immunoglobulin molecule are formed by a rearrangement 
of genes coding for the variable region (V) with genes encoding for a joining 
segment (J) and genes encoding for the constant region (Ck or C2). Heavy 
chains of the immunoglobulin molecule are formed by similar gene rearrange- ^ 
ments which include a fourth region (D for Diversity) which is spaced between 

54 



V and J. The number of genes coding for the D region is still unknown but 
there appears to be a small number (perhaps four) J region genes. If, for a 
heavy chain, any V region gene can combine with any D region gene which can 
in turn associate with any J region gene, the number of combinations possible 
by this shuffling provides a second mechanism for generating antibody diver- 
sity. 

The third process for generating diversity appears to be somatic mutation. 
Upon sequencing a number of monoclonal antibodies reactive with a defined 
simple chemical hapten, it was found that IgM antibodies could be grouped 
into three families based on identity of amino-terminal amino acid sequences. 
A second class of antibody (IgG) reactive to the same hapten and supposedly 
sharing the same V regions as the IgM, consistently showed amino acid sub- 
stitutions not seen in the IgM and which could be explained by a process of 
somatic mutation occurring after V-D-J region combination (2). 

Through these three processes, the immune system maintains a large library 
of combining sites capable of recognizing the determinants on the surface 
of any cancer cell regardless of how or where it develops. Already, infor- 
mation from these types of studies is being integrated into efforts to 
produce antibodies with the most desirable specificities, affinities, sub- 
class type, etc. for application against disease. At the same time, these 
findings provide important leads to research outside the purview of immunology 
but significant to the understanding of how other genes are organized and 
regulated. 

B. Natural Cell Mediated Immunity ; 

Natural cell mediated immunity is an immune effector mechanism unique from 
the standpoint that it represents a cytotoxic capability which exists at a high 
level without previous exposure (i.e. priming) to the target cell. In con- 
trast, responses of bursal equivalent-derived (B) lymphocytes and thymus- 
dependent (T) lymphocytes are generally minimal until after exposure to 
antigen. The role of natural cell mediated immunity against tumors is a par- 
ticularly active research area. 

The cells mediating this type of immunity appear to be rather heterogene- 
ous. For example, in mice one type is called the natural killer (NK) cell 
and a second type is referred to as NC (for natural cell-mediated cytotoxicity) 
distinguishable from NK cells by the type of target cells it is capable of 
lysing, its activity in various mouse strains and other features (3). A 
third type resembling NK in most features but reacting against sarcomas and 
other solid tumors rather than lymphomas is designated NKs. (4). Evidence 
on the presence of cell surface markers indicates that NK cells share some 
common determinants with lymphocytes of the T lineage (5) but the NC cell 
lacks these markers (6) and there is no consensus as to the origin of all 
the various cells which mediate natural cytotoxicity. A feature that is 
characteristic of cells mediating natural cytotoxicity as opposed to other 
forms of cell mediated immunity (such as cytotoxic T cells and antibody-depen- 
dent cytotoxicity by K cells) is their broad target specificity. While NK 
cells are capable of lysing a variety of seemingly unrelated tumors, T cell 

55 



I. 



I 

cytotoxicity and ADCC are specific for either a single tumor or tumors which 
are cross reactive. The broad specificity of NK. cells is evident in cloned 
cell lines (7) and thus does not represent the participation of a hetero- 
geneous population of cells of different specificities. 

The functional properties of NK cells have led to speculation that they 
play a surveillance role in preventing the outgrowth of transformed cells. 
One line of evidence supporting this notion develops from study of the mutant 
"beige" mouse. This animal has very low levels of natural killer activity and 
it has been reported that these deficient animals support more rapid growth 
of transplanted lymphoid tumors than do normal controls (8), thus inferring 
a role of NK cells in limiting t\imor outgrowth. A second approach to this 
question has demonstrated that variants of cell lines selected for their 
ability to produce tiomors in mouse strains with either high or low levels of 
natural cell mediated immunity are respectively resistant or sensitive to 
lysis by NK cells. Again this is consistent with an J^ vivo surveillance 
role by NK cells. 

Although natural cell mediated immunity exists without prior immuniza- 
tion, the level of activity can be modulated. Interferon and substances 
which induce interferon have been shown to be capable of augmenting NK acti- 
vity. In most cases this is demonstrated by a simple increase in killing at 
various effector: target ratios after pretreatment with interferon (9,10,11). 
The mechanism by which this increase occurs appears complex. First, it may 
involve the maturation of pre-NK cells to NK as measured by both the inducti 
of cell surface markers such as LyT-5 and Nk-1 and the ability to become 
cytotoxic (12,13). Secondly, the rate of killing is increased indicating 
a more efficient cytotoxicity function (13). Other soluble mediators can 
also regulate NK activity, i.e. , interleukin-2 or T cell growth factor 
(14). Cellular mechanisms of supression of natural immunity also appear 
to be operative (15). 

The discovery and study of natural cell-mediated immunity has a number of 
ramifications. First, it provides information on a new effector mechanism 
potentially important in anti-tumor immunity. Secondly, this information 
clears up many questions which had arisen in comparing immune responses 
between normal controls and immunized subjects (e.g., the source of high 
"spontaneous" tumor cell killing). Third, the modulation of this activity 
by mediators such as interferon helps explain some of the manifold effects 
of this substance and provides some rational base for therapy. Fourth, 
the broad specificity of natural killer cells may make them very useful in 
adoptive immunotherapy. Underlying this and many other facets of tumor 
immunology is a growing awareness of the complex interactions of the immune 
system as a whole. Because there exists so many regulated interactions, 
biological modulation is likely to pose an extremely complex problem. 

C . Monoclonal Antibodies ; 

The application of somatic cell hybridization to the fusion of a lympho-^ 
cyte with a myeloma cell to produce a hybridoma has provided a major advance " 



56 



in cancer immunology. Hybridomas produce monoclonal antibody molecules which, 
by their very nature, are uniform with respect to binding specificity. Mix- 
tures of antigens unseparable by conventional heterologous antisera are easily 
resolved into individual components by analysis with monoclonal antibodies. 
This provides a new opportunity to dissect the cell surface composition of 
both lymphocytes and the tumor cells they react with. 

For example, Schlossman and colleagues (16,17) have prepared and character- 
ized a series of monoclonal antibodies useful in defining normal human T lym- 
phocyte subclasses and their transformed counterparts. Cell surface antigens 
designated T9 and TIO are found on early thymocytes and most T-cell acute 
lymphatic leukemias appear to be the transformed counterparts of these early 
cells. Cortical thymocytes bear surface antigens TIO, T6, T4 and T5 and some 
T cell-acute lymphatic leukemias and most T cell lymphoblastic lymphomas are 
the transformed counterparts of this stage. The more mature medullary thy- 
mocyte bears the TIO, Tl, T3 and T4 or T5 cell surface antigens and transformed 
counterparts here include a small number of T-cell lymphoblastic lymphomas and 
rare T-cell acute lymphatic leukemias. Peripheral T cells bearing the Tl, T3 
and T4 antigens represent the subpopulation normally involved in the induction 
of immune responses. Most T-cell chronic lymphatic leukemias, cells of the 
Sezary syndrome and cells of Mycosis fungiodes also fall in this category. 
The normal T cell subpopulation capable of suppressing immune responses or of 
exhibiting direct cytotoxicity bear the Tl, T3 and T5 cell surface antigens 
as do the cells in rare cases of T-cell chronic lymphatic leukemia. 

These studies not only shed light on the processes of normal T cell devel- 
opment and function but also provide a subclassification of lymphomas and 
leukemias which may prove useful in diagnosis and in deciding most effective 
treatment. Similar approaches by other laboratories including those of Hansen, 
Levy, Kersey, Springer, Russell and Kaplan are providing additional information 
on cell surface markers for T cells and extend the technique to B cells and 
the monocyte/macrophage. 

Human melanoma is an example of a tumor which is being extensively studied 
from the standpoint of preparing monoclonal antibodies with high relative 
specificities for the tumor which can then be used for studies in biology, 
pathology, diagnosis and potentially even therapy. 

There appears to be several major categories of melanoma associated anti- 
gens. The first is a large glycoprotein of approximately 240,000 daltons 
described by Reisfeld, Ferrone, and colleagues (18). A second smaller gly- 
coprotein (M.W. of 95,000-97,000) has been described by the Hellstroras and 
their colleagues. In addition, this laboratory has developed two very 
sensitive assays for quantitating this antigen on the cell surface which 
should be widely applicable (19). The importance of this kind of technical 
development is that it opens the possibility of diagnostic application when 
there is a quantitative rather than qualitative antigenic difference between 
normal and tumor cells. A third type of glycoprotein antigen which has been 
described is of even lower molecular weight (25,000-40,000) but it seems 
likely that this type represents subunits of larger glycoproteins which are 
in turn relatively specific for melanoma cells and/or is a component of 

57 



f 

histocompatability antigens which are found on melanoma as well as other cells 
(20). A fourth type of melanoma associated antigen which has been described 
by Lloyd, Old and colleagues is a glycolipid (21) which is either identical 
or very similar to the GD3 ganglioside previously shown to exist at very low 
levels in the brain and pigmented areas of the eye. This finding is of in- 
terest in that it focuses attention on a class of biological molecules which 
have not been well studied in terms of tumor antigenicity. Also, it demon- 
strates the applicability of monoclonal antibodies in analyses of non-protein 
antigens. In light of the finding by Steplewski and co-workers that an anti- 
gen quite specific for human colon carcinoma is also a glycolipid, it is 
likely that considerable future attention will be devoted to serological 
analysis of this class of cell surface component. 

The NCI Immunology Program sponsored a workshop on Monoclonal Antibodies 
to Human Melanoma Antigens in March of 1981 to facilitate an exchange of 
information among the major laboratories involved in this area of research. 
This proved to be a useful format in comparing data from the different groups, 
discussing technical details in depth and arranging for an exchange of anti- 
bodies among the participant laboratories. 

From this exchange and comparison, it should be possible to define those 
antibodies most worthy of application in diagnosis and therapy. It is, how- 
ever, equally important to continue the basic studies relating to cell-surface 
antigens on tumor cells. For example, we have yet to learn whether tumor- 
specific antigens exist on human tumors. If the antigens are only tumor- C 
associated (i.e., also found on other tumors or normal cells at some stage of 
development), what is their cellular function and why are they in higher con- 
centration on transformed cells? Also, the ability to work with a defined 
antigen will greatly simplifly the task of studying the induction and regula- 
tion of the immune response against tumor cells. Further, studies on the 
clonality of tumors and on the problem of recurrence versus new tumor develop- 
ment should be greatly facilitated by reference to appropriately defined cell 
surface antigens. 

In an effort to promote the shared use of hybridomas and monoclonal anti- 
bodies, the NCI Immunology Program has supported a Cell Bank and Distribution 
Center which makes a wide variety of cell lines in this and other categories 
available to interested investigators (CB 23886). 



D. Immune Response Genes ; 

Baruj Benacerraf , a grantee in the NCI Immunology Program for the past 
eight years, was awarded the 1980 Nobel Prize in Physiology and Medicine, 
along with George Snell and Jean Dausset. Dr. Benacerraf 's specific ac- 
complishments relate to his studies on the role of the major histocompata- 
bility complex in immune regulation (24). His most recent work continues in 
this area and includes studies on soluble factors bearing antigens determined 
by genes in the Ir region of the major histocompatibility complex. These 
same factors also have regions which are antigenically cross reactive with # 
the antigen binding portion of immunoglobulin molecules. These factors are 

58 



involved in the collaborations which must occur among macrophages, B cells 
and T cells for some types of immune response to occur. 

The future study of these interactions and the genes which govern them will 
be important to our understanding of the functioning of the immune system. 

E. Meeting Support ; 

The following meetings, conferences and workshops were supported by the 
NCI Immunology Program during fiscal year 1981: 

"Mechanisms in Human Cancer Immunology Symposium" - Galveston, Texas 
October 1980 

"Host Defense in Neoplasia" - Plymouth, New Hampshire 
July 1981 

"Mechanisms in Cell Mediated Cytotoxicity" - Carry-Le-Rouet , France 
September, 1981 

"1981 Midwest Immunology Conference" - Minneapolis, Minnesota 
October 1981 

"International Cancer Congress" - Seattle, Washington 
1982 



59 



FISCAL YEAR 1981 

IMMUNOLOGY PROGRAM 

SUMMARY OF GRANTS BY SUBCATEGORY 

(Includes POl, ROl, R23, R13 Grants) 

(Dollars in Thousands) 



f 



Subcategory 



No. of Grants 



Total Costs Awarded 



Myeloma Proteins 20 

Cell Surface Antigens 66 
Cell Surface Determinants of 

Lymphocytes & Macrophages 40 
Humoral Factors other than 

Antibody 36 

Tumor-Related Antibodies 17 
Immunobiology of Sarcomas, 

Carcinomas & Melanomas 10 

Host/Tumor Imraunopathology 22 
Effects of Disease on 

Inmiune Function 51 
Immunotherapy: Mechanisms 

Rather Than Therapeutic Result 8 

Lymphocytes 94 

Monocytes and Macrophages 28 
Malignancies of the Immune 

System (Lymphoma/Leukemia) 33 

Immune Surveillance 32 

Immunotherapy in Animal Models 6 

Bone Marrow Transplantation 7 



$ 2,612 
8,086 

4,207 

3,369 
1,865 

843 

2,422 

5,732 

976 

15,326 

3,572 

3,476 

3,131 

698 

895 



470 



$57,211 



60 



References 



1. Cory, S. and Adams, J.M. , 1980. "Deletions Are Associated with Somatic 
Rearrangement of Immunoglobulin Heavy Genes," Cell , 19:37-51, 

(CA 12421) 

2. Gearhart, P.J. , Johnson, N.D. , Douglas, R . and Hood, L. , 1981. "IgG 
Antibodies to Phosphorylcholine Exhibit More Diversity Than Their IgM 
Counterparts," Nature, 291:29-34, (CA 25507) 

3. Stutman, 0. and Cuttito, M.J. , 1981. "Normal Levels of Natural Cytotoxic 
Cells Against Solid Tumors in NK-Deficient Beige Mice," Nature , 290:254- 
257, (CA 15988) 

4. Burton, R.C., 1980. "Alloantisera Selectively Reactive with NK Cells: 
Characterization and Use in Defining NK Cell Classes", In: Herberman, 
R.B., (ed. ), Natural Cell-Mediated Immunity Against Tumors , Academic 
Press, New York, (CA 17800) 

5. Koo, G.C., Jacobson, J.B., Hammerling, G.J. and Hammerling, U. , 1980. 
"Antigenic Profile of Murine Natural Killer Cells," J. Immuno l. , 125: 
1003-1006, (CA 25416) ~ 

6. Lattime, E.G., Pecoraro, G.A. and Stutman, 0., 1981. "Natural Cytotoxic 
Cells Against Solid Tumors in Mice," III. "A Comparison of Effector 
Cell Antigenic Phenotype and Target Cell Recognition Structures with Those 
of NK Cells," J. Immunol . , 126:2011-2014, (CA 17818) 

7. Dennert, G. , Yogeeswaran, G. and Yamagata, S. , 1981. "Cloned Cell Lines 
with Natural Killer Activity - Specificity, Function and Cell Markers," 
J. Exp . Med . , 153:545-556, (CA 15581). 

8. Karre, K. , Klein, G.O. , Kiessling, R. , Klein, G. and Roder, J.C, 1980. 
"Low Natural rn Vivo Resistance to Syngeneic Leukemias in Natural Killer- 
Deficient Mice," Nature , 284:624-626, (CA 26782) 

9. Orn, A., Gidlund, M. , Ojo, E. , Gronvik, K. , Andersson, J., Wigzell, H. , 
Murgita, R.A. , Sennik, A. and Gresser, I., 1980. "Factors Controlling 
the Augmentation of Natural Killer Cells," In: Herberman, R.B. (ed. ), 
Natural Cell-Mediated Immunity Against Tumors , Academic Press, New York, 
(CB 64033) 

10. Trinchieri, G. , Perussia, B. and Santoli,D. , 1980. "Spontaneous Cell 
Mediated Cytotoxicity: Modulation by Interferon," In: Herberman, R.B. 
(ed. ), Natural Cell Mediated Immunity Against Tumors, Academic Press, 
New York, (CA lOSJl) 



61 



11. Zarling, J.M. , 1980. "Augmentation of Human Natural Killer Cell Activi*^ 
by Purified Interferon and Polyribonucleotides," In: Herberman, R.B. 
(ed. ) , Natural Cell Mediated Immunity Against Tumors , Academic Press, 
New York, (CA 26738) 

12. Minato, N. , Ried, L. , Cantor, H. , Lengyel, P. and Bloom, B.R. , 1980. 
"Mode of Regulation of Natural Killer Cell Activity by Interferon, " 
J. Exp . Med . , 152:124-137, (CB 74147) 

13. Silva, A., Bonavida, B. and Targan, S. , 1980. "Mode of Action of Inter- 
feron-Mediated Modulation of Natural Killer Cytotoxic Activity: Re- 
cruitment of Pre-NK Cells and Enhanced Kinetics of Lysis," ^J. Immunol . , 
125:479-484, (CA 12800) 

14. Henney, C.S., Kuribayashi, K. , Kern, D.E. and Gillis, S. , 1981. 
"Interleuk.in-2 Augments Natural Killer Cell Activity: Natural Killer 
Cell Activity," Nature , 291:335-338, (CA 24537) 

15. Lotzova, E. , 1980. "£. parvum- Mediated Suppression of the Phenomenon 
of Natural Killing and Its Analysis," In: Herberman, R. (ed. ) 
Natural Cell-Mediated Immunity Against Tumors , Academic Press, New 
York, (CA 24537) 

16. Reinberg, E.L. , Kung, P.C, Goldstein, G. , Levey, R.H. and 
Schlossman, S.F. , 1980. "Discrete Stages of Human Intrathymic Differenj 
tiation: Analysis of Normal Thjmiocytes and Leukemic Lymphoblasts of " 
T lineage," Proc . Nat'l . Acad . Sci . (U.S.), 77:1588-1592, (CA 25369) 

17. Schlossman, S., 1981. "Anti-Human T Cell Hybridomas," Workshop on 
Hybridomas in the Diagnosis and Treatment of Cancer . In press. 

(CA 25369) 

18. Galloway, D.R. , Imai, K. , Ferrone, S. and Reisfeld, R.A. , 1981. "Mole- 
cular Profiles of Human Melanoma Associated Antigens," Fed . Proc . , 40: 
231-236, (CA 28420) 

19. Brown, J.P. , Woodbury, R.G. , Hart, C.E., Hellstrom, I. and 
Hellstrom, K.E. , 1981. "Quantitative Analysis of Melanoma-Associated 
Antigen p. 97 in Normal and Neoplastic Tissues," Proc . Nat'l . Acad . Sci . 
(U.S.), 78:539-543 (CA 19149) 



20. Mitchell, K.F., Fuhrer, J.P., Steplewski, Z. and Koprowski, H. , 1980. 
"Biochemical Characterization of Human Melanoma Cell Surfaces: Dissection 
with Monoclonal Antibodies," Proc . Nat'l Acad . Sci . (U.S.), 77:7287-7291, 
(CA 25874) 

21. Dippold, W.G. , Lloyd, K.O., Li, L.T.C., Ikeda, H. , Oettgen, H.F. and 
Old, L.J. , 1980. "Cell Surface Antigens of Human Malignant Melanoma: 
Definition of Six Antigenic Systems with Mouse Monoclonal Antibodies," 
Proc. Nat'l. Acad. Sci. (U.S.), 77:6114-6118, (CA 21445) 



62 



22. Olsson, L. and Kaplan, H.S., 1980. "Human-Human Hybridomas Producing 
Monoclonal Antibodies of Predefined Antigenic Specificity," Proc . Nat ' 1 . 
Acad . Sci . (U.S.), 77:5429-5431, (now supported by CA 29876) 

23. Croce, CM., Linnenback, A., Hall, W. , Steplewski, Z. and Koprowski, H. , 
1980. "Production of Human Hybridomas Secreting Antibodies to Measles 
Virus," Nature , 288:488-489, (CA 23568) 

24. Benacerraf, B. , 1981. "Role of MHC Gene Products in Immune Regulation," 
Science, 212:1229-1238, (CA 14723) 



63 



64 



MYELOMA PROTEINS 



ROl-CA-04946 
Bosma 



Hidden Immunoglobulin Allotypes in Mice 

Institute for Cancer Research 



ROl-CA-08497 
Putnam 

ROl-CA-10056 
Solomon 

ROl-CA-12421 
Adams 



Abnormal Proteins in Multiple Myeloma 

Indiana University Bloomington 

Proteins in Multiple Myeloma and Related Blood Diseases 
University of Tennessee Knoxville 

Structure of Immunoglobulin Messenger RNAs and Genes 
Walter and ELiza Hall Inst. Medical Res. 



ROl-CA-13014 
Beychok 

ROl-CA-13953 
Huang 

ROl-CA-16858 
Morrison 

ROl-CA-19616 

Edmund son 

R01-CA-20G88 
Witz 

ROl-CA-22105 
Warner 

ROl-CA-23885 
Lamm 

ROl-CA-24432 
Haber 

ROl-CA-25049 
Cannon 

ROl-CA-25166 
Levine 

POl-CA-25319 
Beychok 

ROl-CA-25507 
Gearhart 

ROl-CA-25754 
Storb 



Proteins of Plasma Cell Cancers 
Columbia University 

Control of Myeloma Tumor Growth 

Johns Hopkins University 

Genetics and Biochemistry of Myeloma Ig Production 
Columbia University 

Immunoglobulins in Multiple Myeloma and Amyloidosis 
University of Utah 

Tumor-Bound Ig — An Expression of Antitumor Immunity 
Tel Aviv University 

Allotypes of Murine Immunoglobulins 

University of New Mexico Albuquerque 

Secretory Immunoglobulin Studies 
New York University 

Sequence, Shape and Specificity of Antibodies 
Massachusetts General Hospital 

Structure and Genetics of Antibody Variable Regions 
Brandels University 

Defects in Immunoglobulin Production in Mouse Myeloma 
State University of New York Stony Brook 

Synthesis of Human Myeloma Variable Domains in E^. coli 
Columbia University 

B Cell Expression of Immunoglobulin V and C Regions 
Carnegie Institution of Washington, DC 

Control of Immunoglobulin Synthesis 

University of Washington 



65 



ROl-CA-28871 
Green 



Ig Processing by Ljnnphocyte Endoplasmic Reticulum 
St. Louis University 



ROl-CA-29634 Immunoglobulin mRNA and Genes in T Cell Tumors 

Kemp Walter and Eliza Hall Inst. Medical Res. 

ROl-CA-29679 Genetic Analysis of Membrane Immunoglobulin 
Sibley University of Washington 

ROl-CA-31013 Immunoglobulin Genes of Normal and Leukemic Human DNA 
Blattner University of Wisconsin Madison 



CELL SURFACE ANTIGENS 



ROl-CA-12851 Embryonic and Virally Induced Tumor-Cell Membrane Antigens 
Sanders University of Texas Austin 

ROl-CA-13070 Immunity to Human Cancer — Functional Components 
Dawson Duke University 

ROl-CA-13287 Genetic Basis of Antigenic Variation 

Hyman Salk Institute for Biological Studies 

ROl-CA-13844 Isolation of Tumor Antigens of Human Melanoma 
Bystryn New York University 

ROl-CA-14054 Malignant Behavior and Cellular Antigen Expression 
Klein Caroline Institute 

ROl-CA-16069 Immunochemical Characterization of Antigens in Melanoma 
Ferrone Scripps Clinic and Research Foundation 

ROl-CA-17 533 Myelogenous Leukemia- Associated Antigens 

Wust University of Tennessee Knoxville 

ROl-CA-18470 Antigenicity and Tumorigenicity of Somatic Cell Hybrids 
Knowles Wistar Institute of Anatomy and Biology 

ROl-CA-18600 Masking of Antigens at Cancer Cell Surfaces 
Codington Massachusetts General Hospital 

ROl-CA-18609 Biological Role of Alloantigens 

Acton University of Alabama in Birmingham 

ROl-CA-19149 Transplantation Antigenicity of Virus Induced Tumors 
Hellstrom Fred Hutchinson Cancer Research Center 



ROl-CA-19224 
Hakomori 



Relation of Blood Group and Human Tumor Antigens 

Fred Hutchinson Cancer Research Center 



ROl-CA-19304 
Seon 



Human Leukemia and Lymphoma Associated Antigens 
Roswell Park Memorial Institute 



66 



POl-CA-19765 
Old 



Human Cancer Serology 

Sloan Kettering Institute for Cancer Res. 



ROl-CA-20168 
Little 



Neoplastic and Normal Cell Thymus-Leukemia Antigens 
Jewish Hospital of St. Louis 



ROl-CA-21223 
Levy 

ROl-CA-21279 
Cooper 



ROl-CA-22540 
Springer 

ROl-CA-22674 
Coggin 



ROl-CA-23568 
Croce 



Antitumor Antibodies Generated In Vitro 
Stanford University 

Isolation and Characterization of Fc Receptors 



ROl-CA-21445 Antigens of Human Malignant Melanoma 

Lloyd Sloan Kettering Institute for Cancer Res. 

POl-CA-22507 Immunogenetics of the Major Histocompatibility Complex 

Dupont Sloan Kettering Institute for Cancer Res. 



Human Cancer Relation of MN and Precursor Structures 
Evanston Hospital 

Characterization of Fetal Antigens in Tumors 
University of South Alabama 



ROl-CA-22794 Human Neuroblastoma Cell Surface Antigens 

Seeger University of California Los Angeles 

POl-CA-23115 New Immunologic Approaches to Lymphoid Neoplasms 

Frenkel University of Texas Hlth. Sci. Ctr. Dallas 

ROl-CA-23404 Serologic Screening for Contagion of Human Sarcomas 

Hirshaut Sloan Kettering Institute for Cancer Res. 



Immunoresponse to Human Surface Antigens 

Wistar Institute of Anatomy and Biology 



ROl-CA-24024 In Vitro Generation of Immunity to Human Colon Cancer 

Tom University of Texas Hlth. Sci. Ctr. Houston 

ROl-CA-24910 Oncofetal Antigens in Embryogenesis and Tumor Growth 
Rosenberg University of Maryland Bait. Co. Campus 



ROl-CA-25139 
Warren 



Study of Group 5 Antigens in Hematologic Malignancies 
Fred Hutchinson Cancer Research Center 



ROl-CA-25154 Tumor-Specific Immunity and Histocompatibility Complex 

Chauvenet University of Texas Hlth. Sci. Ctr. San Antonio 

ROl-CA-25171 Cell Surface Antigens of Murine Tumors 

Callahan Scripps Clinic and Research Foundation 



ROl-CA-25558 

Hellstrom 



Cell Surface Antigens of Chemically Induced Sarcomas 
Fred Hutchinson Cancer Research Center 



67 



ROl-CA-25852 
Merrick 



Isolation and Characterization of Heterophile Antigens 
State University of New York, at Buffalo 



POl-CA-25874 Human Melanoma and Tumor Specific Monoclonal Antibodies 
Koprowski Wistar Institute of Anatomy and Biology 



ROl-CA-25910 
Leung 

ROl-CA-26184 
Lloyd 

ROl-CA-26321 
Allison 



Production of Antibodies to Tumor Associated Antigens 

Rutgers, The State University New Brunswick 

Antigens of Human Ovarian Tumors 

Sloan Kettering Institute for Cancer Res. 

Cell-Surface Antigens of Murine Tumors 

University of Texas System Cancer Center 



ROl-CA-26456 Cell Surface Antigens in Murine Leukemia 

Callahan Scripps Clinic and Research Foundation 



ROl-CA-26479 
Fuji 

R23-CA-26584 
Woodbury 



Immune Functions of Tumor Cell Variants 

Roswell Park Memorial Institute 

The Molecular Nature of Tumor Antigens 

Fred Hutchinson Cancer Research Center 



ROl-CA-26891 Surface Antigens of Rat Hepatocellular Carcinomas 

Allison University of Texas System Cancer Center 

ROl-CA-27124 Molecular Approaches to Human Colon Cancer 

Kahan University of Texas Hlth. Sci. Ctr. Houston 



ROl-CA-27471 

Shinitzky 



Modulation of Cellular Responses by Membrane Fluidity 
Weizmann Institute of Science 



ROl-CA-27534 Nucleolar Antigens of Human Cancer Cells 
Busch Baylor College of Medicine 

ROl-CA-27628 Tumor-Specific and Tumor-Associated Antigens 

Milgrom State University of New York at Buffalo 



ROl-CA-27841 
Brown 



Antigens of Chemically Transformed Mouse Fibroblasts 
Fred Hutchinson Cancer Research Center 



POl-CA-28166 Molecular Immunology and Pathobiology of Neoplasia 
Edgington Scripps Clinic and Research Foundation 



ROl-CA-28212 
Mind en 



Antisera for Human Tumor-Associated Antigens 

National Jewish Hospital & Research Ctr. 



ROl-CA-28230 Swine Melanoma Antigens: Isolation and Evaluation 
Hook University of Missouri Columbia 

ROl-CA-28253 Immune Response to Syngeneic Leukemic B Cell Antigens 
Ricardo University of Tenn. Center Health Scien. 



68 



ROl-CA-28420 
Reisfeld 



ROl-CA-28461 
De Leo 



ROl-CA-28564 
Carey 



ROl-CA-28775 
Anderson 



Molecular Profile of Human Melanoma Antigens 

Scripps Clinic and Research Foundation 



ROl-CA-28448 Forssman Antigen/Antibody and Human Adenocarcinoma 

Levine Sloan Kettering Institute for Cancer Res. 



Cell Surface Antigens of Mouse Sarcomas 

Sloan Kettering Institute for Cancer Res. 



R23-CA-28526 Analysis of Human Leukemia Cells with Hybridomas 

Le Bien University of Minnesota of Minneapolis-St . Paul 



Human Squamous Cell Carcinoma: Culture and Serology 
University of Michigan 



ROl-CA-28564 Human Squamous Cell Carcinoma: Culture and Serology 
Carey University of Michigan 

ROl-CA-28619 Purification of Human Sarcoma Heterophile Antigens 

Hirshaut Sloan Kettering Institute for Cancer Res. 

ROl-CA-28732 Studies of Immune Complexes in Patients with Leukemia 
Williams University of New Mexico Albuquerque 



Cross-Reacting Antigens on Spermatozoa and Tumors 
Medical Research Foundation of Oregon 



ROl-CA-29216 Characterization of a Tumor Antigen on Leukemia Cells 
Carver Medical College of Georgia 



ROl-CA-29516 
Wright 



ROl-CA-29662 
Klock 



Human Antibodies to Melanoma 

Pacific Northwest Research Foundation 



ROl-CA-29534 Oral Immunopathology: Oral Squamous Cell Carcinoma 
Eskinazi University of Southern California 

R23-CA-29539 Role of Glycolipids in Immune Cell Function 
Young University of Virginia 



Complex Carbohydrate Chemistry in Leukocytes 

University of California San Francisco 



ROl-CA-29863 

Michaelson 



Immunochemical Genetics of Murine Alloantigens 
New York University 



ROl-CA-29886 Tumor-Host Associated Immunological Specificities 
De Witt University of Utah 



ROl-CA-29897 

Pellegrino 

ROl-CA-29989 
Pollack 



Antigenic Profile of Human Leukemic Cells 

Scripps Clinic and Research Foundation 

HLA Alloantigens on Cultured Human Tumor Cell Lines 

Sloan Kettering Institute for Cancer Res. 



69 



ROl-CA-30209 Immunochemical Studies of Gastrointestinal Cancer 
Alpert Baylor College of Medicine 

ROl-CA-30266 Membrane Antigen Organization in Tumor Immunity 
Gooding Emory University 

ROl-CA-30501 Isolation and Characterization of the Common ALL Antigen 
Mills City of Hope National Medical Center 

ROl-CA-30561 Tumor Associated Antigens of UV-Induced Tumors 

Spellman University of New Mexico Albuquerque 



CELL SURFACE DETERMINANTS OF LYMPHOCYTES AND MACROPHAGES 



ROl-CA-04681 

Herzenberg 

ROl-CA-10097 

Wettstein 

ROl-CA-14061 
Gilmour 

ROl-CA-15146 
Gasser 

ROl-CA-15318 
Lalezari 

ROl-CA-16071 

Pellegrino 

ROl-CA-17680 
Collins 

ROl-CA-18640 
Silvers 



Genetic Studies with Mammalian Cells 
Stanford University 

Cellular Antigens of Normal and Malignant Rat Tissues 
Wistar Institute of Anatomy and Biology 

Chicken Lymphocyte Alloantigens and Viral Oncogenesis 
New York University 

Genetic Control of the Immune Response 
University of Pennsylvania 

Neutrophil Antigens; Pathophysiology and Chemistry 
Montefiore Hospital and Medical Center 

Urine as Source of Human Cell Surface Markers 

Scripps Clinic and Research Foundation 

Genetics, the Lymphocyte and Tumor Regression 
University of New Hampshire 

Behavior of Weak Transplantation Antigens 
University of Pennsylvania 



ROl-CA-18659 
Gill 

ROl-CA-18734 
Jones 

ROl-CA-20107 

Winchester 

ROl-CA-20473 
Boyse 



Chemical Genetic and Cellular Aspects of Immunogenicity 
University of Pittsburgh 

Immunologic Studies Related to Malignancy 

University of Colorado Hlth. Sciences Ctr. 

B Cell Alloantigens, Molecular Basis and Disease Aspects 
Rockefeller University 

Immunogenetics of the TLA Region of Chromosome 17 

Sloan Kettering Institute for Cancer Res. 



70 



ROl-CA-20500 Structural and Serological Studies on la Antigens 
Cullen Washington University 

ROl-CA-20531 Genetic Analysis of Normal and Malignant Ljnnphocytes 
Yunis Sidney Farber Cancer Institute 

ROl-CA-20820 Structural Studies of the Products of the H-2 Complex 
Freed Johns Hopkins University 



POl-CA-21112 
Osserman 



Dyscrasias of Plasma Cells and Macrophages 
Columbia University 



ROl-CA-21651 
ArtzT 



Teratocarcinoma and Embryonal Tumors: Surface Antigens 
Sloan Kettering Institute for Cancer Res. 



ROl-CA-22080 Cell Surface Antigens of Lymphocytes and Tumor Cells 
McKenzie University of Melbourne 

ROl-CA-22131 Immunogenetics of Ly Systems 

Boyse Sloan Kettering Institute for Cancer Res. 

ROl-CA-22662 Genetics and Function of Murine la Antigens 

Frelinger University of Southern California 

ROl-CA-23026 The Major Histocompatibility Complex of the Rabbit 
Tissot University of Illinois Medical Center 

ROl-CA-23027 Immunogenetic Mapping of the Cell Surface 

Flaherty New York State Department of Health 

ROl-CA-23030 Structure Studies of la Alloantigens 

Silver Scripps Clinic and Research Foundation 

ROl-CA-23469 Cells Involved in Spontaneous Regression of Tumors 
Yang University of Connecticut Storrs 

ROl-CA-23677 New Method for Separating Leucocyte Subpopulations 
Parker University of Southern California 

ROl-CA-24433 Structures of Histocompatibility-2 Membrane Antigens 
Sears University of California Santa Barbara 

ROl-CA-24437 Expression of T Linnphocyte Differentiation Antigens 
Esselman Michigan State University 

ROl-CA-24473 Genetics and Functions of (H-2 Linked) I Region 
David Mayo Foundation 



ROl-CA-25038 
Cramer 



Major Histocompatibility Complex in the Wild Rat 
University of Pittsburgh 



ROl-CA-25041 Murine T Lymphocyte Receptors for Alloantigens 
Neefe Georgetown University 



71 



ROl-CA-25044 Surface IgM of Malignant Lymphocytes and Plasma Cells 
Hickman Jewish Hospital of St. Louis 

ROl-CA-25893 Cell Surface Molecules: Hematopoietic Differentiation 
Hyman Salk Institute for Biological Studies 

ROl-CA-26297 Primary Structure of MHC I Region Associated Antigens 
McKean Mayo Foundation 

ROl-CA-27063 Genetics of Cytotoxicity to Virus Modification in Man 
Dubey Sidney Farber Cancer Institute 

ROl-CA-27547 Chemistry of Tumoricidal Macrophage Surface Antigens 
Springer Harvard University 

ROl-CA-27824 Tumor Membrane Composition and Immune Function 
Whisnant Duke University 



ROl-CA-27955 
Williams 



Role of the H-2K Gene in Hybrid Resistance 
Northwestern University 



R23-CA-28271 Character of B Lymphocyte Differentiation Antigens 
Ades Medical University of South Carolina 



ROl-CA-28992 Membrane Lectins on Normal and Neoplastic Lymphocytes 
Decker Medical University of South Carolina 

ROl-CA-29111 Expression of H-2 Antigens on SJL/J Tumors 
Beisel Wayne State University 

ROl-CA-29194 Somatic Cell Genetics of Cell Surface Antigens 
Rajan Yeshiva University 



ROl-CA-29548 
Hansen 



Differentiation Antigens on Human Ljnnphocytes 

Pacific Northwest Research Foundation 



ROl-CA-29657 

Haran-Ghera 



Genetic Control in Leukemogenesis 

Weizmann Institute of Science 



R23-CA-29738 Macrophage Membrane and Immunomodulators 

Mitchell University of Southern California 



ROl-CA-29979 
Yamazaki 



Immunogenetics of Self- Identification 

Monell Chemical Senses Center 



ROl-CA-30654 Regulation of Immune Responses by Fc Portion of Antibody 
Morgan Scripps Clinic and Research Foundation 

ROl-CA-31799 Chemistry of Tumoricidal Macrophage Surface Antigens 
Springer Sidney Farber Cancer Institute 



72 



HUMORAL FACTORS OTHER THAN ANTIBODY 



ROl-CA-01786 Human Blood and Tissue Proteins 

Deutsch University of Wisconsin Madison 

R01-CA-07191 T Lymphocytes and Lymphotoxin in Tumor Immunity 

Rosenau University of California San Francisco 

ROl-CA-15129 A Serum Immunosuppressive Factor in Cancer 
Moolten Boston University 

ROl-CA-17643 Regulation of T-Cell Proliferation and Differentiation 
Smith Dartmouth College 

ROl-CA-18893- Killing of Cancer Cells by BCG Activated Cells 

Warfel Sloan Kettering Institute for Cancer Res. 



ROl-CA-19148 

Hellstrom 



Lymphocyte Allogeneic Inhibition and Tumor Immunity 
Fred Hutchinson Cancer Research Center 



ROl-CA-19529 

Valentine 



Cell-Mediated Immunity in Humans: Mechanisms and Uses 
New York. University 



ROl-CA-19721 Human Interferon Production for Antitumor Studies 
Tan University of Calgary 

ROl-CA-20819 Phagocytic Cells: Regulation, Dysfunction and Disease 
Van Epps University of New Mexico 



ROl-CA-23681 
Turner 

ROl-CA-24441 
Mayer 

ROl-CA-24447 
Kolb 

ROl-CA-24476 
Incefy 

ROl-CA-24916 

Sundharadas 



Factors Affecting Phagocyte Responses to Lipids 
Duke University 

Biochemical Studies of Lymphokines and Related Agents 
Johns Hopkins University 

Complement Attack Proteins as Lymphocyte Surface AGS 
University of Texas San Antonio 

T-Lymphocyte Differentiation 

Sloan Kettering Institute for Cancer Res. 

Studies of a Tumor Factor That Affects Macrophages 
University of Wisconsin Madison 



ROl-CA-24974 

Goldstein 



Chemical and Immunological Characteristics of Thymosin 
George Washington University 



ROl-CA-25388 Complement and Immune Complexes in Lymphosarcoma 

Day Sloan Kettering Institute for Cancer Res. 

ROl-CA-25750 Structure-Function Relations of Immuno regulatory Protein 
Miller University of Chicago 



73 



ROl-CA-25756 Regulation of Cell Growth by 5 '-Methylthioadenosine 
Ferro Oregon State University 

ROl-CA-25943 Immuno bio chemistry of Macrophage-Derived Factors 

Elgert Virginia Polytechnic Inst, and St. Univ. 

ROl-CA-26019 Isolation of Macrophage Agglutination Factor 

Godfrey State University of New York Stony Brook 

ROl-CA-26051 Lymphokines Formed In Vivo: Role in Tumor Suppression 
Moulton University of Texas Med. Br. Galveston 

ROl-CA-26143 Control of Complement-Mediated Tumor Cell Cytolysis 

Lint Rush-Presbyterian-St . Luke's Medical Ctr. 

ROl-CA-26462 Role of Macrophage Arginase in Tumor Immunity 

Reiss University of Colorado Hlth. Sciences Ctr. 

ROl-CA-26504 Regulation of Granulocyte and Macrophage Production 
Stanley Yeshiva University 

ROl-CA-26842 Biological Functions of Alpha 2 Macroglobulin 

Stein-Streilein University of Texas Hlth. Sci. Ctr. Dallas 



ROl-CA-27452 
Sundsmo 

R23-CA-27565 
Sharma 

ROl-CA-27629 
Pa que 

ROl-CA-27701 
Ladisch 

ROl-CA-27903 
Epstein 

ROl-CA-28123 
Huang 

ROl-CA-28419 
Gillis 

ROl-CA-28471 
Dvorak 

R23-CA-29828 
Bernhard 

ROl-CA-30015 

Mortensen 



Complement Membrane Attack Proteins Binding and Biosynthesis 
Trudeau Institute 

Tumor Cell Killing by a Soluble Macrophage Product 
Palo Alto Medical Research Foundation 

Tumor Immune RNA: A Biochemical Characterization 

University of Texas Hlth. Sci. Ctr. San Antonio 

Human Immunoregulatory Gangliosides 

University of California Los Angeles 

Interferon as a Mediator of Cellular Immunity 

University of California San Francisco 

The Induction of Human Interferon in C-10 Cells 
Johns Hopkins University 

Control of Normal and Leukemic T-Cell Proliferation 
Fred Hutchinson Cancer Research Center 

Biology of Solid Tumor Growth and Immune Rejection 
Beth Israel Hospital 

Lymphokine-Directed Monocyte-Macrophage Cytotoxicity 
University of Virginia Charlottesville 

C-Reactive Protein Regulation of Tumor Immunity 
Ohio State University 



74 



ROl-CA-30114 Complement Membrane Attack Proteins 

Sundsmo Scripps Clinic and Research Foundation 

ROl-CA-30651 Monocyte Tissue Factor: In Vivo and In Vitro Modulation 
Edwards University of Connecticut Health Center 

R23-CA-30669 Growth and Differentiation of Mast Cells and T Cells 

Yung Sloan Kettering Institute for Cancer Res. 

R23-CA-30988 Tumor Specific Helper Factor(s) 

Mathews Loyola University Medical Center 



TUMOR RELATED ANTIBODIES 



ROl-CA-15064 
Chu 



Immunochemical Studies on Carcinogenic Mycotoxins 
University of Wisconsin Madison 



ROl-CA-15333 Rheumatoid Factor and Tumor-Host Interaction 
Twomey Baylor College of Medicine 

ROl-CA-16328 Polyamine Radioimmunoassay Studies in Cancer 

Campbell University of Oregon Hlth. Sciences Ctr. 

ROl-CA-20045 Antibody Mediated Cell-Cell Interactions 
Phillips-Quagli New York University 

ROl-CA-20075 Antibody Affinity in Immune Response and Tolerance 
Siskind Cornell University Medical Center 



ROl-CA-23028 
Richards 



Molecular Studies of the Immune Response 

California Institute of Technology 



ROl-CA-23967 Radiolabeled Antibody Tumor Localization 

Buchsbaum University of Minnesota of Minneapolis-St • Paul 

ROl-CA-24329 Monoclonal Antibodies to Human Cell Surface Antigens 
Ferrone Scripps Clinic and Research Foundation 

ROl-CA-25958 Organ Specific Tumor Localizing Antibody 

Bale Georgia Institute of Technology 

ROl-CA-26882 Antibody Responses of Tumor Bearers to Their Tumors 
Klein University of Florida 

ROl-CA-28149 Immunotherapy of a Mouse B Cell Leukemia (BCLl) 

Vitetta -University of Texas Hlth. Sci. Ctr. Dallas 

ROl-CA-29876 Human Hybridoma Antibodies in Neoplastic Disease 
Kaplan Stanford University 

ROl-CA-29885 Tumor Cells Escape from Immune Effector Mechanisms 

Kuo University of Tenn. Center Health Scien. 



75 



ROl-CA-29889 Targeting Antibody-Toxin Conjugates to Leukemia Cells 
Houston University of Kansas Lawrence 

ROl-CA-30313 New Approaches to the Therapy of a B Cell Leukemia 
Slavin Hadassah University Hospital 

ROl-CA-30647 In Vitro Synthesis of Human Antibodies to Oncofetal AG 
Irie University of California Davis 

ROl-CA-30663 Antibody-Directed Tumor Specific Chimeric Toxins 
Collier University of California Los Angeles 

ROl-CA-30990 Monoclonal Antibodies to Human Lung Carcinoma Antigens 
Bell Washington University 



IMMUNOBIOLOGY OF SARCOMAS, CARCINOMAS, AND MELANOMAS 



ROl-CA-14462 Properties of Lymphoid Tumor Cells In Vivo and In Vitro 
Thorbecke New York University 

ROl-CA-19753 Mixed Leukocyte Tumor Reaction in Syngeneic Systems 
Bonavida University of California Los Angeles 

ROl-CA-20364 Immunodiagnosis of Melanoma 
Seigler Duke University 

ROl-CA-23891 Immunobiology of ASV Induced Primary Brain Tumors 
Roszman University of Kentucky 

ROl-CA-25214 Immunological Studies of Herpes Induced Fibrosarcoma 
Gusdon Wake Forest University 

ROl-CA-26226 Childhood Neuroblastoma: Antigenic Characteristics 
Casper Medical College of Wisconsin 

ROl-CA-27134 Immunobiology of Human Malignant Melanoma 

Pickering Louisiana State Univ. Med. Ctr. Shreveport 

ROl-CA-28311 Immunobiology of Murine Primary Rous Sarcoma 

Haughton University of North Carolina Chapel Hill 

ROl-CA-28611 Teratocarcinoma Tumor Associated Fetal Embryonic Antigen 
De Wolf Sidney Farber Cancer Institute 

R01-CA-29007 Melanoma Surface Antigens and Cytotoxic T Cells 

Oettgen Sloan Kettering Institute for Cancer Res. 

ROl-CA-30461 Clonal Analysis of Cellular Immune Response in Melanoma 
Mukherji University of Connecticut Health Center 



76 



HOST-TUMOR IMMUNOPATHOLOGY 



POl-CA-16835 Studies of Monoclonal Gammopathies in Humans 
Kyle Mayo Foundation 

ROl-CA-16869 Oncornea Cell Membrane Antigens and Its Therapeutics 
Theilen University of California Davis 

ROl-CA-17800 Tumor Immunology 

Winn Massachusetts General Hospital 

ROl-CA-18995 Studies of Tumor Dormancy and Emergence 
Wheelock Thomas Jefferson University 

ROl-CA-20044 Transplantation Immunology 

Winn Massachusetts General Hospital 

ROl-CA-22869 Significance of Tumor Leucocytic Infiltrates 

Kreider Pennsylvania State Univ. Hershey Med. Ctr. 

ROl-CA-23024 Allograft Rejection and Enhancement 
Tilney Harvard University 

ROl-CA-23477 Immunologic Inhibition of Tumor Growth 
Forbes Vanderbilt University 

RGl-CA-23679 Cell Mediated Hyperacute Rejection 
Eichwald University of Utah 

ROl-CA-24196 Ultraviolet Carcinogenesis and Immunity 
Fortner Kansas State University 

ROl-CA-24215 Mechanisms of Metastasis 

Kim Roswell Park Memorial Institute 



ROl-CA-24726 
Roholt 



Immune Mechanisms Involved in Tumor Metastasis 
Roswell Park Memorial Institute 



ROl-CA-24728 Tumor Induced Changes in Lymphatic Tissues 
Anderson Johns Hopkins University 

ROl-CA-25965 Study of Micrometastasized Stable Murine Tumor Clones 
Ghanta University of Alabama in Birmingham 

R23-CA-27893 Bence Jones Protein Properties and Tubulotoxicity 
Chander New York Medical College 



R01-CA-28060 
Frost 



Immunobiology Metastasis 

University of California Irvine 



ROl-CA-28139 
Feldman 



The Immunobiology of Tumor Metastasis 

Weizmann Institute of Science 



77 



R23-CA-30110 
Galli 



Vascular Damage in Skin Allograft and Tumor Rejection 
Beth Israel Hospital 



ROl-CA-30169 Bone Allografts for Surgical Oncology 
Friedlaender Yale University 

ROl-CA-30565 Growth Factor (s) in Nodular Sclerosing Hodgkin's Disease 
Newcom University of Oregon Hlth. Sciences Ctr. 



ROl-CA-31199 
Russell 

R13-CA-31821 
Mi rand 



Macrophage-Mediated Injury Causing Tumor Regression 
University of Florida 

International Cancer Congress (UICC) 



EFFECTS OF DISEASE ON IMMUNE FUNCTION 



ROl-CA-10267 
Rosse 



Immunological Lysis in Neoplastic Disease 
Duke University 



ROl-CA-14300 Mechanism of Immunosuppression by Plasmacytomas 
Havas Temple University 

POl-CA-15147 Leukemia — Lymphoma Program 

Balcerzak Ohio State University 

ROl-CA-15334 Cellular Mechanisms in Tumor-Specific Immunity 
Smith • University of Florida 

ROl-CA-15462 Cell Interactions in Tumor Immunity 
Argyris Upstate Medical Center 



ROl-CA-15585 

Zolla-Pazner 



A Soluble Mediator of Tumor- Induced Immunosuppression 
New York University 



ROl-CA-17273 The Immune Response to Virally Determined Tumor Antigens 
Lamon University of Alabama in Birmingham 



ROl-CA-17818 
Stutman 



Tumor Immunity and Tumor-Host Interactions 

Sloan Kettering Institute for Cancer Res. 



ROl-CA-18149 Autoimmunity and Neoplasia in New Zealand Black Mice 
Siegel University of Oregon Hlth. Sciences Ctr. 

ROl-CA-18185 Hereditarily Athymic — Asplenic Mice: Tumor Growth 
Lozzio University of Tennessee Knoxville 

ROl-CA-18234 Immunobiology of Primary Intracranial Tumors 
Roszman University of Kentucky 



POl-CA-19267 Clinical Immunobiology Program Project 

Oettgen Memorial Hospital for Cancer-Allied Diseases 

ROl-CA-20543 Antigen-Antibody Complexes in Cancer Patients' Sera 
Rossen Baylor College of Medicine 

ROl-CA-20816 The Pathogenesis of Autoimmunity in New Zealand Mice 
Gershwin University of California Davis 



ROl-CA-20920 
Prehn 

ROl-CA-23217 
Lynch 

ROl-CA-23372 
Laing 

ROl-CA-23500 
Bockman 

ROl-CA-23648 
Haskill 

ROl-CA-23709 
Adler 

ROl-CA-23882 
Neef e 

ROl-CA-24244 
Brody 

ROl-CA-24429 

Winkelstein 

ROl-CA-24511 
Mayers 

ROl-CA-24698 
Humphrey 

ROl-CA-24725 
Lucas 

ROl-CA-24873 

Bankhurst 

ROl-CA-24901 
Outzen 

ROl-CA-25072 
Raich 



Mechanisms of Carcinogenesis 

Jackson Laboratory 

Immunologic Regulation of Myeloma Cell Growth 
Washington University 

Studies on Soluble Antigens on MTV Induced Mammary Tumor 
Howard University 

Prostaglandins and Immunosuppression in Cancer 

Sloan Kettering Institute for Cancer Res. 

Immunity to Human Ovarian Tumors and Chemotherapy 

University of North Carolina Chapel Hill 

Induction of Tolerance and Suppressor Cells In Vitro 
St. Jude Children's Research Hospital 

Defective Self-Recognition as a Cause of Cancer 
Georgetown University 

Studies of the Immunobiology of B-16 Melanoma 
Downstate Medical Center 

Immunosuppressants and Lymphocyte Function 
Montefiore Hospital 

Auto-regulation of Immune Response by Anti-idiotype 
New York State Department of Health 

Arginine Pulse Labeling of Plasma Cell Neoplasms 
Johns Hopkins University 

Factors Negating Response to Mammary Adenocarcinoma 
Stanford University 

Immunosuppression in Cancer Patients 

University of New Mexico Albuquerque 

Immunomodulation of Oncogenesis 
Jackson Laboratory 

Suppressor T-Cells in Malignant Lymphoma 
West Virginia University 



79 



ROl-CA-25183 
Wood 

ROl-CA-25746 

Fundenberg 

R23-CA-25932 
La t time 

R23-CA-26116 
Nelson 

ROl-CA-2 6141 
Yu 

ROl-CA-26169 
Bose 

ROl-CA-26268 
Dube 

ROl-CA-26447 
Hoover 

ROl-CA-26760 
Weston 

ROl-CA-26861 

Giovanella 

ROl-CA-27168 
Osband 

ROl-CA-27390 
S pence 

ROl-CA-28167 
Ekstedt 

R23-CA-28433 
Ostenson 

ROl-CA-29200 
Guerry 

ROl-CA-29752 
Devens 

ROl-CA-29906 
Kadish 

ROl-CA-30020 

Aisenberg 



Immunologic Factors in Central Nervous System Tumors 

University of Kansas Col. Hlth. Sci. & Hosp. 

Genetic Mechanisms in Immunologic Deficiency States 
Medical University of South Carolina 

Thymic Antigens, Tolerance and Self -Recognition 

Sloan Kettering Institute for Cancer Res. 

Antigen-Specific Suppression of Antitumor Immunity 
Fred Hutchinson Cancer Research Center 

Cell-Mediated Cytotoxicity in Children with Neoplasms 
University of California San Diego 

Immunosuppression During Acute Avian Leukemia 
University of Texas Austin 

Anti-II Antibodies and II Antigens in Carcinomas 
Evanston Hospital 

Pathogenesis of Preleukemic Aplastic Anemia 
Ohio State University 

Monocyte-Lymphocyte Interactions in Mycosis Fungoides 

University of Colorado Hlth. Sciences Center 

Determination of Malignant Potential of Cultured Cells 
Stehlin Foundation for Cancer Research 

Histamine Receptor Positive T-Cells in Cancer 
University Hospital 

Ethylnitrosourea-Induced Rat Gliomas 
University of Washington 

Tumor Enhancement in Lectin Treated Mice 
Northwestern University 

Identification of AG Specific Suppressor Cells in Man 
Fred Hutchinson Cancer Research Center 

Autologous Immunity to Human Cultured Melanoma 
University of Pennsylvania 

Immune Response Modulation by Tumor Promoter 

University of California Riverside 

Mechanisms of Immunoregulation in Human Cancer 
Yeshiva University 

The Cell Surface Phenotype of Malignant Lymphoma 
Massachusetts General Hospital 



80 



ROl-CA-30088 Synergy of Tumor Chemotherapy and Host Immunity 

Dray University of Illinois Medical Center 

R23-CA-30160 Cytotoxic and Suppressor Cells in the Chicken 
Chi East Tennessee State University 

ROl-CA-30187 Regulation of Cell-Mediated Cytotoxicity Mechanisms 
Bloom University of California Los Angeles 

ROl-CA-30457 Immunoregulation of Human Tumor Growth in Nude Mice 
Koros University of Pittsburgh 

ROl-CA-30660 Immuno regulatory Dysfunctions in Non-Hodgkin's Lymphoma 
Keller Medical College of Wisconsin 

ROl-CA-30920 Immunosuppression in Murine Chronic Lymphocytic Leukemia 

Uhr University of Texas Health Science Center Dallas 

ROl-CA-30933 Immunomodulatory Factors in Head and Neck Cancer 
Veltri West Virginia University 

ROl-CA-31226 Tolerance and Immunity to Avian RNA Tumor Viruses and VI 
Meyers Mayo Foundation 

ROl-CA-31336 Antigen Evasion as a Tumor Escape Mechanism 
Stackpole New York Medical College 



ROl-CA-31837 
Prehn 



Santa Clara Valley Medical Center 



IMMUNOTHERAPY-MECHANISM RATHER THAN THERAPEUTIC RESULT 



ROl-CA-11605 Immunological Reactivity in Special Circumstances 

Simmons University of Minnesota at Minneapolis 

ROl-CA-18047 Irradiation and Marrow Transplantation in Large Animals 
Thomas University of Washington 

ROl-CA-26138 Immune Testing in Lung Cancer During Immunotherapy 
Harris Rush University 

ROl-CA-26162 Mechanism of Nanaase-enhanced Tumor Immunogenicity 
Pincus SRI International 

ROl-CA-26738 Cellular Immunity to Tumors 

Zarling University of Minnesota of Minneapolis-St . Paul 

ROl-CA-27625 Hybrid Tumor Cell Immunotherapy 

McCune University of Rochester 



81 



ROl-CA-28441 Extracorporeal Immunoadsorbents in Inmuno therapy 

Tennan Baylor College of Medicine 



ROl-CA-28941 
Deeg 



ROl-CA-31787 
Thomas 



Resistance and Sensitization-Role of Lymphocyte Subsets 
Fred Hutchinson Cancer Research Center 



ROl-CA-29328 Control of Graf t-Versus-Host Disease 

Parkman Sidney Farber Cancer Institute 



Irradiation and Marrow Transplantation in Large Animals 
Fred Hutchinson Cancer Research Center 



LYMPHOCYTES 



ROl-CA-03367 Immunogenetic Resistance to Lymphoma- Leukemia 
Trentin Baylor College of Medicine 



ROl-CA-08593 
Gershon 



Immune Responses to Tumor Grafts 
Yale University 



POl-CA-12800 Immune Functions and Cancer 

Fahey University of California Los Angeles 

ROl-CA-12844 Controls of Proliferation Specific for Leukemias 

Cudkowicz State University of New York at Buffalo 

ROl-CA-13396 Immunogenesis from Bone Marrow Cells 

Miller Michigan State University 

ROl-CA-14049 Cell-Mediated Immunity to Ascites Tumors 
Amos Duke University 

ROl-CA-14216 Characterization of Lymphoid Populations in Cancer 
Gershon Yale University 



POl-CA-14723 

Benacerraf 



Study of Experimental Cancer Immunology 
Harvard University 



POl-CA-15822 Immunobiology of Normal and Neoplastic Lymphocytes 
Wilson University of Pennsylvania 



POl-CA-16247 
Lawrence 



Immunologic Resistance to Cancer 
New York University 



ROl-CA-16271 Regulatory Signals in Cytotoxic T-Cell Development 

Faanes Sloan Kettering Institute for Cancer Res. 

ROl-CA-16367 Lymphocytes: Gene Expression and Activation Events 
Sell University of California San Diego 



82 



POl-CA-16673 
Cooper 

ROl-CA-16885 
Ruddle 

ROl-CA-17013 
Golub 

POl-CA-17404 
Choi 

ROl-CA-17531 
Manning 

ROl-CA-17673 
Hoffmann 

ROl-CA-17733 

Trowbridge 

ROl-CA-19170 

Bernstein 



Cell Differentiation Studies in Cancer Immunobiology 
University of Alabama in Birmingham 

Propagation of Thymus-De rived Lymphocyte Lines 
Yale University 

Lymphocyte Sensitization to Tumor Antigens In Vitro 
University of California Los Angeles 

Immunobiology, Immunodeficiency, and Cancer 

Sloan Kettering Institute for Cancer Res. 

Mechanism and Uses of Anti-Ig Immunosuppression 
University of Wisconsin Madison 

Regulation of Immunity by Antibody and B-Cells 

Sloan Kettering Institute for Cancer Res. 

L3miphocyte Antigens: Structure, Function and Synthesis 
Salk Institute for Biological Studies 

Mechanisms of BCG-Mediated Suppression of Tumor Growth 
Fred Hutchinson Cancer Research Center 



ROl-CA-19334 
Dennert 

ROl-CA-20105 

Wohlgemuth 

ROl-CA-20169 
Click 

ROl-CA-20823 
Rosse 

ROl-CA-21401 
Kumar 

POl-CA-21825 
Grey 



Antigen Receptor of Continuous T Killer Cell Line 
Salk Institute for Biological Studies 

Information from Immunological Reaction Tables 
University of Maine at Orono 

Long-Lived Lymphocytes and Secretory Bursal Cells 
Mississippi State University 

Ljnnphocyte Production and Traffic in the Bone Marrow 
University of Washington 

Studies of Isolated Marrow-Dependent M Cells 
Boston University 

Self-Nonself Discrimination and Tumor Recognition 

National Jewish Hospital & Research Ctr. 



ROl-CA-22093 Functions of Atypical Ljmiphocytes 

Hutt-Fletcher University of North Carolina Chapel Hill 



ROl-CA-22126 
Daynes 



Ultraviolet Light Radiation and Immunoregulation 
University of Utah 



ROl-CA-22241 Lymphocyte Subsets in Immune-Deficiency States 

Scheid Sloan Kettering Institute for Cancer Res. 



ROl-CA-22268 Genetic Aspects of Tumor Host Immune Relationship 
Warner University of New Mexico Albuquerque 



ROl-CA-22544 Lymphocyte Responses to Syngeneic Antigens 
Ponzio Northwestern University 

ROl-CA-22677 Pathobiology of Myeloma and Anti-idiotypic Immunity 
Schreiber University of Chicago 

ROl-CA-22786 Receptor Dynamics and Normal/Tumor Cell Function 
Bankert Roswell Park Memorial Institute 

ROl-CA-22845 Immune Response to Modified Self and Tumor Antigens 
Scott Duke University 

ROl-CA-23025 Comparative Transplantation Immunogenetics 

Hildemann University of California Los Angeles 

ROl-CA-23262 DNA Polymerases in Normal and Leukemic Lymphoid Cells 

Bollum U.S. Uniformed Services Univ. of Hlth. Sci. 

ROl-CA-23354 Natural Tumor Cell Killing in Humans 
Koren Duke University 

ROl-CA-23593 Response of Leukocytes to Human Tumor Cells 

Sanderson University of Colorado Hlth. Sciences Ctr. 

ROl-CA-24335 Lectin-Dependent Cell-Mediated Cytotoxicity 
Laux University of Rhode Island 

ROl-CA-24338 In Vitro Studies of Normal and Neoplastic Lymphocytes 
Fu Rockefeller University 

ROl-CA-24368 Immunobiology of Early Lymphocyte Development 
Sato Harvard University 

ROl-CA-24431 Cellular and Structural Basis of Immunological Tolerance 
Benjamin University of Virginia 

ROl-CA-24436 Receptor Function in Lymphocyte Differentiation 
Wofsy University of California Berkeley 

ROl-CA-24438 Genetic Control of Allotype Expression of Human Ig 
Litwin Cornell University Medical Center 

ROl-CA-24442 Chemical Basis for Receptor Recognition of Lysozymes 
Sercarz University of California Los Angeles 

ROl-CA-24450 T-Cell Receptor and Effector Molecules 

Redelman University of California San Diego 

ROl-CA-24472 Development of Thymic L3rmphocytes 
Basch New York University 



ROl-CA-24537 
Henney 



Murine Effector Cells 

Fred Hutchinson Cancer Research Center 



84 



ROl-CA-24607 HLA Restricted Suppressor T Cells of Mixed Lymphocytes 
Engleman Stanford University 

ROl-CA-25054 Cellular Mechanisms Regulating Antibody Production 
Mullen University of Missouri Columbia 

ROl-CA-25250 Natural Killer Cells: Genetic Control and Role 
Klein Caroline Institute 



ROl-CA-25253 
Bankert 



Immune regulatory Network Probed by Cell Hybridization 
Roswell Park Memorial Institute 



ROl-CA-25416 
Koo 

ROl-CA-25508 

Hammerling 

ROl-CA-25583 
Lopez 

ROl-CA-25612 
Plate 

ROl-CA-25738 
Scheid 



Immunogenetics of NK-1+ Natural Killer Cells 

Sloan Kettering Institute for Cancer Res. 

Ontogenetic Relationship of CR+ and CR- B Lymphocytes 
Sloan Kettering Institute for Cancer Res. 

Cell Mediated Immunity in Mouse Mammary Tumor Models 
University of Miami 

Immunological Effects on Tumor Growth, and Rejection 
Rush-Presbyterian-St Lukes Medical Ctr. 

T Cell Differentiation: Molecular Mechanisms 

Sloan Kettering Institute for Cancer Res. 



ROl-CA-25747 Histocompatibility Antigens in Defined Membranes 
Gilmer Florida State University 

POl-CA-25803 Control of Normal and Abnormal Cell Development 

Katz Scripps Clinic and Research Foundation 

ROl-CA-26084 Interactions Between Tumor Cells and T Lymphocytes 
Hale Wake Forest University 

ROl-CA-26257 Modulation of Lymphocyte Cytolysis of Tumor Cells 
Fuson University of Tennessee Knoxville 

ROl-CA-26284 Regulation of Adenosine Deaminase in Human Cells 
Daddona University of Michigan 

ROl-CA-26467 Effector and Suppressor Mechanisms of Tumor Immunity 
Stout Brandeis University 

ROl-CA-26480 Antitumor Activity of Tumor-Bearer Lymphoid Cells 
Dray University of Illinois Medical Center 

ROl-CA-26695 Antigen-Specific T-Cell Clones: Generation and Analysis 
Cantor Sidney Farber Cancer Institute 

ROl-CA-26713 Genetics and Regulation of Cell-Mediated Cytotoxicity 
Clark University of Washington 



R23-CA-27115 Characterization of Natural Killer Cells 
Tai University of New Mexico 



R23-CA-27552 
Green 



Cytotoxic T Cells to Syngeneic MuLV+ Tumors 

Fred Hutchinson Cancer Research Center 



ROl-CA-27691 
Ozer 



Immunoregulation by T Cell Subsets in Myeloma and CLL 
Roswell Park Memorial Institute 



ROl-CA-27772 In Vitro Immunization to Human Tumor Cells 

Sharma University of California Irvine 

ROl-CA-27854 Cell Surface Carbohydrate and Ljnnphocyte Interactions 
Bell University of Rochester 



ROl-CA-27 915 
Fanger 



Antibody Dependent Cell Cytotoxicity Reactions 
Case Western Reserve University 



ROl-CA-28099 F.A.C.S. of Immunologic Components 
Amos Duke University 

ROl-CA-28196 Proteinases of Human Natural Killer Cells 

Hudig University of California San Diego 

ROl-CA-28332 In Situ Anti-tumor Immunity and Effects of Radiation 
Lord University of Rochester 

ROl-CA-28533 Mechanisms of Tumor Destruction by Immune Effectors 
Russell Washington University 

ROl-CA-28708 Immunoregulation of Myeloma Cell Differentiation 
Rohrer University of South Alabama 



POl-CA-28900 
Eisen 



Control of Antigen-Specific T Cell Responses 

Massachusetts Institute of Technology 



ROl-CA-28936 Immunoregulation in Autoimmunity and Malignant Disease 
Haynes Duke University 

ROl-CA-29208 Fc Receptor-Bearing T Lymphocytes in Murine Myeloma 
Lynch Washington University 

R23-CA-29224 Cytotoxic T Ljnnphocyte Lines to Murine Plasmacytomas 
Giorgi University of New Mexico Albuquerque 



ROl-CA-29282 
Waksal 



Prothymocyte Maturation and Function 
Tufts University 



ROl-CA-29594 

Catsimpoolas 

POl-CA-29606 
Gershon 



Lymphoid Cell Purine Nucleoside Phosphorylase 
Boston University 

Immunoregulation: Studies on T Cells and Their Products 
Yale University 



86 



ROl-CA-29635 Analysis of Human T Ljnnphocyte Subsets Grown In Vitro 
Pauly Roswell Park Memorial Institute 

R23-CA-29803 Cytotoxic Cell Responses to Non-H2 Antigens 

MacPhail Sloan Kettering Institute for Cancer Res. 

ROl-CA-30147 Genetic Markers, Leukemogenesis and Thymic Function 
Gottlieb University of Texas Austin 

R23-CA-30183 Bone Marrow Cytotoxic Precursor T Cells 

Klimpel University of Texas Med. Br. Galveston 

R23-CA-30188 Autorecognition and Immunoreactivity 
Scheffel Marquette University 

ROl-CA-30280 T-Lymphocyte Regulated Tumor Cell Killing by Neutrophils 
Weisbart University of California Los Angeles 

ROl-CA-30972 Marrow Prostaglandins and T-Cell Differentiation 

Bockman Sloan Kettering Institute for Cancer Res. 

R13-CA-31141 "Host Defense in Neoplasia" (1981 Gordon Conference) 
Dvorak Beth Israel Hospital 

R13-CA-31191 Int'l Workshop in Mechanisms in Cell-Mediated Cytotoxicity 
Clark University of California Los Angeles 

R13-CA-31451 Midwest Autumn Immunology Conference 
Schmidtke Eli Lilly and Company 

R23-CA-31591 Regulation of Human B Cell Proliferation 
Yen University of Iowa 



MONOCYTES AND MACROPHAGES 



ROl-CA-14113 Tumor Defense by Platelets and Macrophages 
Shin Johns Hopkins University 

ROl-CA-15236 Macrophage Recognition and Tumor Cell Interactions 
Schreiber University of Pennsylvania 

ROl-CA-16652 Macrophage Functions in Tumorigenesis 

Walker St. Jude Children's Research Hospital 

ROl-CA-16784 Tumoricidal Effects of Macrophages: Pathologic Study 
Adams Duke University 

ROl-CA-18672 The Role of Macrophage Subclasses in Tumor Immunity 
Fishman St. Jude Children's Research Hospital 

ROl-CA-19052 Development and Function of Activated Macrophages 

Moore Sloan Kettering Institute for Cancer Res. 



87 



ROl-CA-20822 Cell Interaction and the Clotting System 

Colvin Massachusetts General Hospital 

ROl-CA-21225 Macrophage Activation by Lymphocyte Mediators 
Remold-0 'Donnel Center for BLood Research 



ROl-CA-22090 
Na than 

ROl-CA-23503 
Edelson 

ROl-CA-24686 
Morahan 

ROl-CA-25052 

Niederhuber 

R23-CA-26158 
Stux 

ROl-CA-26824 

Mantovani 

ROl-CA-26846 
Musson 

ROl-CA-26996 
Fi shman 



Mononuclear Leukocytes in Tumor Immunity 
Rockefeller University 

Mechanisms of Macrophage Antitumor Activation 

Children's Hospital Medical Center 

Macrophage Extrinsic Activity Vs Viruses 

Virginia Commonwealth University 

Immune Responses In Vitro-H-2 (IR) Locus Function 
University of Michigan 

The Immunogenetics of Macrophage Suppression in Man 
Sidney Farber Cancer Institute 

Mononuclear Phagocytes in Human Ovarian Carcinoma 

Mario Negri Institute Pharmacologic Res. 

Mechanism of Human Monocyte Differentiation 

National Jewish Hospital & Research Ctr. 

Characterization and Functional Studies of "A" Cells 
St. Jude Children's Research Hospital 



ROl-CA-27070 Macrophage Tumor Cell Killing and Red Cell Catabolism 
Weinberg Duke University 

ROl-CA-27523 Macrophages and Regulation of Tumor Growth 
Evans Jackson Laboratory 

ROl-CA-27694 Cytotoxic Macrophages Activation and Target Recognition 
Tompkins University of Illinois Urbana-Champaign 

ROl-CA-28308 Differentiation and Anti-tumor Activity of Macrophages 
Kaplan Virginia Commonwealth University 

R23-CA-28935 Macrophage Mediated Tximor Cytotoxicity 

Cameron Medical University of South Carolina 

ROl-CA-29266 Characterization of Monocyte Subsets in Blood 
Weiner University of Florida 

R23-CA-29333 Metabolic Events Related to Macrophage Activation by MVE 
Klykken Virginia Commonwealth University 

ROl-CA-29336 Macrophage-Mediated Cytotoxicity of Tumor Targets 
Erickson University of California Davis 



88 



POl-CA-29589 Macrophage Activation: Development and Regulation 
Adams Duke University 

POl-CA-30198 Human Mononuclear Leukocytes in Cancer 
Silverstein Rockefeller University 

R23-CA-30631 Targets of a Leukosis Virus Infection 
Price Trenton State College 

ROl-CA-31202 Monoclonal Antibody Depletion of Macrophages In Vivo 
Russell University of Florida 



MALIGNANCIES OF THE IMMUNE SYSTEM (LYMPHOMA/LEUKEMIA) 



ROl-CA-03367 
Trentin 



Immunogenetic Resistance to Lymphoma-Leukemia 
Baylor College of Medicine 



ROl-CA-08975 Human Leukemia Associated Antigens 
Metzgar Duke University 

ROl-CA-10018 Experimental Model of Malignant Lymphoma 

Schwartz New England Medical Center Hospital 

ROl-CA-12 779 Leukocyte Regulatory Mechanisms 

Nowell University of Pennsylvania 

ROl-CA-13 701 Mechanisms of Immunity in Leukemia 
Murphy University of Michigan 



ROl-CA-14413 
Minowada 



Study of Human Lymphatic Neoplasia 

Roswell Park Memorial Institute 



ROl-CA-15472 Immunity to Myeloma Tumors 

Eisen Massachusetts Institute of Technology 



ROl-CA-17 276 
Pressman 



Membrane Antigens from Normal and Leukemic Lymphocytes 
Roswell Park Memorial Institute 



ROl-CA-18602 
Casper 



Immunocompetent Cells in Acute Lymphocytic Leukemia 
Medical College of Wisconsin 



ROl-CA-20499 
Edelson 



Immunobiology of Cutaneous T Cell Lymphomas 
Columbia University 



ROl-CA-21062 NK Cells in Resistance to Marrow Transplantation 

Lotzova University of Texas System Cancer Center 



ROl-CA-21900 
Duffey 



Protective T-Cell Subpopulation Responses in Leukemia 

University of Texas Hlth. Sci. Ctr. San Antonio 



ROl-CA-22948 Gene Mechanisms in Lymphoma and Lymphoprolif eration 
Murphy Jackson Laboratory 



89 



ROl-CA-23770 Antigen Induced Lymphoma of Mice 

Haughton University of North Carolina Chapel Hill 

ROl-CA-24679 Extra-nodal Ljnnphomas: Immunology and Ultrastructure 
Knowles Columbia University 

ROl-CA-24950 A Thymus Determined Mechanism of Leukemia Resistance ' 
Datta Tufts University 

ROl-CA-25097 Differentiation of Inmune System: Cell Surface Antigens 
Kersey University of Minnesota at Minneapolis 



ROl-CA-25369 

Schlossman 



Human Leukemia Antigens: Isolation and Characterization 
Sidney Farber Cancer Institute 



ROl-CA-25391 Immunological Control of Dormant Leukemia 
Dietz Michigan Cancer Foundation 

ROl-CA-25411 SJL Mice L3nnphomagenesis as a Model of B-Cell Lymphoma 
Ford University of Texas System Cancer Center 



ROl-CA-25613 
Ross 



Membrane Components of Normal and Leukemic Leukocytes 
University of North Carolina Chapel Hill 



ROl-CA-25873 Membrane Proteins of Human Leukemias and Lymphomas 

Humphreys University of Massachusetts Medical School 

ROl-CA-26369 T-MICG and N-MICG in Lymphoid Malignancy 
Hauptman Thomas Jefferson University 

ROl-CA-26945 Detecting Paul-Bunnell Antigen in Lymphoma and Leukemia 
Fjelde New York State Department of Health 



ROl-CA-27416 

Mohanakumar 



Characterization of New Human lA and Leukemia Antigen 
Virginia Commonwealth University 



R23-CA-27542 T Cell Subsets and Marek's Disease Viral Oncogenesis 
Fredericksen New York University 

ROl-CA-27690 Multiple Cell Marker Analysis in Hematopoietic Tumors 
Koziner Sloan Kettering Institute for Cancer Res. 

ROl-CA-27826 Manipulation of Antitumor Immunity In Vitro 

Bach University of Minnesota of Minneapolis-St . Paul 

ROl-CA-27942 Role of the Spleen in the Growth of a B Cell Leukemia 
Strober Stanford University 

ROl-CA-28416 Receptors for Immunoglobulin on Human Lymphoma-Leukemia 
Rudders New England Medical Center Hospital 

ROl-CA-28504 T Cell Growth and Differentiation in Leukemia 

Chiao Sloan Kettering Institute for Cancer Res. 



90 



ROl-CA-28746 Cell Surface Antigens in Neoplastic and Autoimmune Disease 
Fox Scripps Clinic and Research Foundation 

ROl-CA-29655 Cytogenetic Studies of Human Myeloma 

Mackenzie University of California Davis 

ROl-CA-29964 UNC-CH Immunocytomas 

Haughton University of North Carolina Chapel Hill 

R23-CA-30895 Differentiation of Murine B Cell Lymphomas 

Lanier University of New Mexico Albuquerque 

ROl-CA-31792 Immunobiology of Viral Leukemia and Its 

Bennett University of Texas Hlth. Sci. Ctr. Dallas 



IMMUNE SURVEILLANCE 



ROl-CA-11898 
Bigner 

ROl-CA-12461 
Wheelock 

ROl-CA-13339 
Weksler 

ROl-CA-15 988 
Stutman 

ROl-CA-16136 
Sell 

ROl-CA-19754 
Cohn 

ROl-CA-20408 
Shultz 

ROl-CA-20833 

Trinchieri 

ROl-CA-21360 
Spitalny 

ROl-CA-22517 
Nermann 

ROl-CA-22720 
Long 

ROl-CA-23809 
Saksela 



Brain Tumors: Virological and Immunological Studies 
Duke University 

Suppression of Established Leukemia Virus Infections 
Thomas Jefferson University 

Host Defense Against Lymphoblastic Leukemia 

Cornell University Medical Center 

Immune Surveillance and Cancer 

Sloan Kettering Institute for Cancer Res. 

The Role of Rabbit Lymphoid Cells in Tumor Immunity 
University of California San Diego 

Immunoselection and Cancer: A Problem in Evolution 
Salk Institute for Biological Studies 

Immunodeficiency and Tumorigenesis 
Jackson Laboratory 

Cell-Mediated Cytotoxicity in Humans 

Wistar Institute of Anatomy and Biology 

Subversion of Immune Mechanisms in Malignant Tumors 
Trudeau Institute 

Monocyte Function in Neoplasia 

University of Florida 

H-2 Complex and Susceptibility to Mammary Tumor Virus 
Sidney Farber Cancer Institute 

Natural and Tissue-Specific Immunity to Human Neoplasms 
University of Helsinki 



91 



ROl-CA-24443 Immunology of Germfree Genetically Th5Tnusless Mice 
Reed University of Montana 

ROl-CA-24497 Immunogenetic Analysis of Rous Sarcoma Development 
McBride Baylor College of Medicine 



ROl-CA-24608 
Grant 



Tumor Immunity and Leukemia 

Harvard University 



ROl-CA-25165 Specific Immunity and Prognosis in Breast Cancer 
Black New York Medical College 



ROl-CA-25384 
Kim 



Natural and Antibody-Dependent Cellular Cytotoxicity 
Sloan Kettering Institute for Cancer Res. 



ROl-CA-25686 Epstein-Barr Virus: Effects on Human Lymphocytes 

Sumaya University of Texas Hlth. Sci. Ctr. San Antonio 

ROl-CA-25917 Cellular and Genetic Aspects of Antitumor Immunity 
Daynes University of Utah 



R23-CA-25944 
Johnson 



Relationship of Leukemogenesis and Immune Responsiveness 
Jackson Laboratory 



ROl-CA-26144 
Powell 



Leukocyte Adherence Inhibition Assay in Human Cancer 
Case Western Reserve University 



ROl-CA-26344 Autologous Lymphocyte Reactions and Immune Surveillance 
Weksler Cornell University Medical Center 

ROl-CA-26752 Relevance and Functions of Natural Killer Cells 
Wigzell University of Uppsala 

ROl-CA-26782 In Vivo Role of Natural Killer Cells 
Kiessling Caroline Institute 

ROl-CA-26799 Immunological Reactivities and MTV Infections 

Tagliabue Mario Negri Institute Pharmacologic Res. 

ROl-CA-26942 H-2 Linked Resistance to Tumor: Effectors and Targets 
Fuji New York State Department of Health 

ROl-CA-27599 Genetic Control of Resistance and Immunity to P815 
Williams Northwestern University 



ROl-CA-28231 
Carlson 



H-2 Associated Natural Resistance 
Jackson Laboratory 



ROl-CA-28834 Basophil/Mast Cell Function in the Control of Cancer 
Dvorak Beth Israel Hospital 

ROl-CA-29355 T-Cell Nonresponsiveness in Gross Virus-Infected Mice 
Blank University of Pennsylvania 



92 



ROl-CA-29910 Human NK Cells, Interf eron(s) and Leukemia/Lymphoma 
Pattengale University of Southern California 



ROl-CA-30115 
Babcock 



Immune Reactivity to Primary Sarcomas in Mice 

University of Texas Hlth. Sci. Ctr. Houston 



ROl-CA-31836 
Prehn 



Santa Clara Valley Medical Center 



IMMUNOTHERAPY IN ANIMAL MODELS 



ROl-CA-15446 Antigen-Antibody Interactions in Neoplastic Disease 
Minden National Jewish Hospital & Research Ctr. 



ROl-CA-16642 
North 



Immunological Basis of Tumor Regression 
Trudeau Institute 



R23-CA-25668 
Leech 



Induction of Genetically Tailored Immune Imbalance 

Louisiana State Univ. Med. Ctr. New Orleans 



ROl-CA-27794 
North 



Mechanisms of Endotoxin-Induced Tumor Regression 
Trudeau Institute 



ROl-CA-30303 Selective Stimulation of Cell Mediated Cancer Immunity 
Hunter Emory University 

R23-CA-30686 Ex Vivo Immunosorption as Tumor Therapy 

Jones Sloan Kettering Institute for Cancer Res. 



BONE MARROW TRANSPLANTATION 



ROl-CA-20044 
Winn 



Transplantation Immunology 

Massachusetts General Hospital 



R01-CA-20484 
Bortin 



Specific Adoptive Immunotherapy of AKR Leukemia 
Mount Sinai Medical Center 



R01-CA-23602 Bone Marrow Transplantation After Ly Subset Elimination 
Krown Sloan Kettering Institute for Cancer Res. 

ROl-CA-23678 Tolerance of Bone Marrow Grafts 

Click University of Minnesota of Minneapolis-St . Paul 



ROl-CA-26245 
Truitt 



Mechanisms of Successful Adoptive Immunotherapy 
Mount Sinai Medical Center 



ROl-CA-28701 

Beschorner 



Chronic Graf t-Versus-Host Disease in Radiation Chimera 
Johns Hopkins University 



93 



ROl-CA-29592 Active Specific Immunotherapy in Man: A Murine Model 

y^han University of Texas Hlth. Sci. Ctr. Houston 



UNCLASSIFIED 



R23-CA-29155 Stress and Human Cell-Mediated Immunity 
Locke Beth Israel Hospital 



94 



CONTRACT RESEARCH SUMliARY 

Title: Induction of Colonic Tumors in Guinea Pigs 

Principal Investigator: Dr. Gary L. Cockerell 

Performing Organization: Cornell University 

City and State: Ithaca, NY 

Contract Number: NOl-CP-65744 

Starting Date: 9/30/76 Expiration Date: 3/29/81 

Goal: To produce carcinomas of the colon in approximately 700 NIH guinea 
pigs and subsequently test the immunotherapeutic effect of BCG vaccines 
versus other modalities of treatment. 

Approach: N-methyl-nitrosourea is administered by intrarectal installation 
to produce colonic carcinomas. Following tumor induction, laparotomies are 
performed, and the tumorous portion of bowel is (1) injected intralesionally 
with BCG vaccines, (2) surgically resected and a diverting colostomy estab- 
lished, or (3) located but not otherwise treated. Animals are then allowed 
to recover from surgery and are observed until death. 

Progress: Two sets of 240 guinea pigs each were exposed to the intrarectal 
administration of methylnitrosourea (MNU) for 14 to 35 weeks to induce colonic 
adenocarcinomas. At 24, 28 or 35 weeks post start of MNU administration, 
groups of guinea pigs were treated either by intratumoral injection of BCG 
or surgical excision of tumors and compared to non-treated animals. Most 
guinea pigs died within one year of therapy and there was no difference in 
survival rate between treated and non-treated groups. The leukocyte migra- 
tion inhibition assay, lymphocyte blast transformation assay with variations 
reaction, and macrophage cytotoxicity assays have been used to evaluate the 
immune reactivity of MNU-exposed guinea pigs. Macrophage cytotoxicity 
has been the only assay to yield useable results but there are yet insuffi- 
cient therapeutic modalities. A third set of 240 guinea pigs has been exposed 
to lesser amounts of MNU to derive carcinogen dose response data. These data 
will make it possible to examine the effects of immunotherapy of surgical 
treatment on tumors which arise in a smaller percent of MNU-exposed animals 
and which might therefore be less malignant. 



Significance to Cancer Research: The guinea pig colon carcinoma model will 
serve as a useful system to test various methods of tumor treatment, since it 
has now been shown that the guinea pig colon tolerates intramural BCG injection 
and is surgically resectable. 

Project Officer: Ms. Judith Whalen 
Program: Immunology Section 
FY 81 Funds: 



95 



CONTRACT RESEARCH SUMMARY 



Title: 



Long-term Tumor-specific Cytotoxic Lymphocytes in Treatment of Mouse 
Leukemia 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-74141 
9/30/77 



Dr. Kendall A. Smith 
Dartmouth Medical School 
Hanover, NH 



Expiration Date: 9/29/81 



Goal: To evaluate the usefulness of tumor-specific cytotoxic lymphocytes 
maintained in continuous culture for the prevention and treatment of mouse 
leukemia. 

Approach: Attempts will be made to (1) demonstrate the tumor-antigen reacti- 
vity of human and murine cytotoxic T-cell lines; and (2) test in. vivo the 
immunoprophylactic and immunotherapeutic value of murine cytotoxic T cells 
(CTLL) maintained in continuous proliferative culture. 

Progress: Preliminary experiments show that murine CTLL (1 x 107 cells) may 
be effective in prolonging survival when administered three days after a lethal 
IV dose (1 X 103 cells) of leukemic cells. Progress was made toward the con- 
struction of a murine model to test the use of CTLL for the eradication of 
leukemia cells prior to autologous bone marrow infusion in mice which receiveB 
lethal irradiation. The investigators report that the supplementation of cul- 
tures comprised of normal unprimed murine spleen cells and syngeneic tumor 
cells with partially purified, lectin-free TCGF leads to the generation of a 
primary i^ vitro cytotoxic response. Based on his data, the investigator has 
constructed a model which depicts the biological role of TCGF in the T-cell 
immune response. 



Significance to Cancer Research: The ability to cultivate large quantities 
of tumor-specific cytotoxic lymphocytes which retard _in vivo tumor growth may 
provide a new approach to cancer treatment. The successful culture of differ- 
entiated functional T cells for extended periods offers the promise of new 
understanding of the mechanisms of cellular immunity and of immunodef icient 
and hyperimmune disease states. 

Project Officer: Harriet L.G. Gordon, M.D. 
Program: Immunology Section 
FY 81 Funds: 



96 



i 



CONTRACT RESEARCH SUMMARY 

Title: Characterization of Antigen-binding T-cell Receptors 

Principal Investigator: Dr. P.H. Krammer 

Performing Organization: Institute for Immunology 

City and State: Heidelberg, Germany 

Contract Number: NOl-CB-74179 

Starting Date: 9/30/77 Expiration Date: 3/31/81 

Goal: To test for the expression of alloantigen receptors on murine 
T-cell tumors. 

Approach: Use AKR/J lymphomas; develop sensitive assay systems for alloanti- 
gen binding T-cell receptors; obtain sufficient quantities of receptor 
material from positive tumors to perform biochemical analyses. 

Progress: Assays for prolif lerative and cytotoxic T cell responses have been 
standardized. Utilization of these assays permits rapid screening of a large 
niimber of continuous lines of normal cytotoxic T cells for receptor expres- 
sion. The specificity of two idiotype systems have been established, using 
this assay and other techniques. Anti-idiotypic antisera made in 
(AKRxC57Bl/6)Fl mice against AKR anti-C57Bl/6 MLR activated T cell blasts 
recognizes AKR T cell receptors for C57B1/6 alloantigens. Anti-idiotypic 
antisera made in syngeneic AKR mice against AKR anti-TNP activated T cell 
blasts (containing AKR anti-TNP H-2 restricted cytotoxic T cells) recognizes 
idiotypes on T cell receptors of AKR cells cytotoxic for syngeneic TNP coupled 
target cells. Over fifty AKR T cell lines have been established in tissue 
culture for several months, growing in the presence of interleukin 2 (T cell 
growth factor). These lines have been cloned by limiting dilution and are 
derived from primary anti-TNP activated lymphocytes. The contractor spent the 
remaining contract period increasing the number of growing clones, testing and 
selecting them for anti-TNP specificity. 



Significance to Cancer Research: T cells play a decisive role in tumor de- 
fense. The recognition process of the T cell in general and the recognition 
of tumor targets in particular are not understood at all. One clue to this 
understanding is the biochemical nature of the T-cell antigen recognition 
system. 

Project Officer: Ms. Judith M. Whalen 
Program: Immunology Section 
FY 81 Funds: 



97 



CONTRACT RESEARCH SUMMARY ^ 

Title: Genetic Control of Susceptibility to Tumors 

Principal Investigator: Dr. Chen K. Chai 

Performing Organization: Jackson Laboratory 

City and State: Bar Harbor, ME 

Contract Number: NOl-CB-74181 

Starting Date: 9/30/77 Expiration Date: 2/28/81 

Goal: Elucidate the reasons why AY mice are more susceptible to lung tumors 
than aa_ mice. 

Approach: Study the carcinogenic responses of the homozygotes and hetero- 
zygotes for the different yellow and non-yellow alleles at the agouti locus. 

Progress: A study was made on the response to urethane for lung tumor pro- 
duction in mice of 20 genotypes for 6 alleles at the agouti locus. The mean 
number of lung tumors was the highest in the A^AW mice; the next highest was 
the Aya_ mice. Means for these two genotypic groups are significantly greater 
than those of the remaining 18 genotypes. All yellow genes at the agouti 
locus showed no significant effect in lung tumor development. Mean numbers 
of lung tumors for the yellow mice in the hybrid generations of each C57BL/6 
X A/J crosses are not greater than the means of the non-yellow mice, indicatit r 
no nonallelic interactions of the yellow genes with the genes conferring sus" 
ceptibility in the A/J. However, a trimodel distribution of lung tumors 
showed in the F2 of the C57BL/6 (A^^) x A/J, suggesting the presence of a major 
gene affecting lung tumor susceptibility in the A/J. Since the primary objec- 
tive of this project was to study the effect of different yellow genes and 
specific allelic combinations for lung tumor development, it is of interest 
to note that mice of Ay in combination with A^ (the wild type), but not A^Y 
or A^ mutant), produced duced on the average the greatest greatest number 
of tumors. The agouti locus, as a complex regulating locus, consists possibly 
of a number of repeating nucleotide sequences. The yellow genes may represent 
different deficiences, the Ay being the most deficient. Under these circum- 
stances, more frequent unequal intragenic crossing over in the AyA" may occur 
than in other allelic combinations, thus resulting in more somatic cell muta- 
tions. Mutated cells are more susceptible to mutagens or are inferior in 
repairing ability. As gene regulation in growth and development in eukaryotes 
is most poorly understood, this interpretation must remain highly speculative. 
The present results, however, point to the importance of mutation of a regu- 
lating locus and possible genetic mechanisms involved in neoplastic development. 

Significance to Cancer Research: To determine the role that a gene, or a 
specific genetic locus, plays in cancer development. The results of this 
investigation would specifically concern the mutation theory of neoplasm. 

Project Officer: Ms. Judith M. Whalen 

Program: Immunology Section ^ 

FY 81 Funds: W 



98 



CONTRACT RESEARCH SUMMARY 

Title: Studies on Macrophage Activation 

Principal Investigator: Dr. Emil Skamene 

Performing Organization: Montreal General Hospital 

City and State: Montreal, Canada 

Contract Number: NOl-CB-84269 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal: Study the interactions of T cells or their products with macrophages 
that lead to activation of macrophages for reaction against tumor cells. 

Approach: Investigate the identity and function of cell populations that 
control macrophage activation in the development of antilistereal resistance 
and determine the cellular level of phenotypic expression of genetically 
controlled high or low levels of antilistereal immunity. 

Progress: Detailed autoradiographic studies dealing with the kinetics of 
monocytopoiesis clearly show that one step being regulated is the cell cycle 
time (Tq) of promonocyotes, this being significantly shorter in mice of the 
genetically-resistant (Lr r ) strain. This leads to increased mononuclear 
phagocyte cellularity not only in the bone marrow, but in every macrophage 
compartment so far examined. High numbers and decreased life span (Tl/2) 
of monocytes in the circulation of infected (Lri^) BIO. A strain mice are 
also evident, indicating efficient delivery of macrophage precursors to 
infective foci. The expression of such a change in monocyte kinetics can be 
documented well in a model situation of the macrophage inflammatory reaction 
in response to local irritants, injected intraperitoneally. Linkage analysis 
on segregating populations has provided the formal evidence that the L£ gene, 
regulates such macrophage response. Three possible approaches to effect a 
change from the sensitive to resistant phenotype exist: (a) replacing 
the animal with the putative Lr^ gene product, (b) elevating the mono- 
cytopoiesis of the LrS animal experimentally to the level in Lr^ animals, and 
(c) by-passing the need for prompt macrophage inflammatory response by effec- 
tive activation of the resident macrophage population. The latter two are 
being actively examined. Splenectomy proved to be an effective maneuver to 
enhance the macrophage response in the genetically susceptible Lr ^ host, 
probably by macrophage proliferation in the foci of infection in the liver. 
Another type of intervention capable of reversing the Lr^ phenotype into 
the resistant one, was chronic stimulation with BCG. Interestingly, the 
BCG treatment, although very effective in macrophage activation for listeri- 
cidal activity in the Lr ^ A mice, proved ineffective in correcting the 
macrophage tumoricidal defect of these animals. 



Project Officer: Ms. Judith M. Whalen 
Program: Immunology Program 
FY 81 Funds: 



99 



CONTRACT RESEARCH SUMMARY 
Title: International Bone Marrow Transplant Registry 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-AI/CB-02648 
5/1/80 



Dr. Mortimer Bortin 

Mount Sinai Medical Center 

Milwaukee, WI 



Expiration Date: 6/30/83 



Goal: To aid in improving the success rate of bone marrow transplantation 
as applied for the treatment of patients with a variety of otherwise incur- 
able diseases including aplastic anemia and hematologic malignancies. 

Approach: Maintain a statistical center for the collection, organization and 
analysis of clinical data provided by transplant teams throughout the world; 
and disseminate the results of analyses of pooled data to bone marrow teams 
and to the scientific community. 

Progress: Bone marrow transplant teams throughout the world reported de- 
tailed data in a uniform fashion to the Resistry at an unprecedented rate 
during the first year of this Contract. The Statistical Center received and 
processed reports of 209 new patients. As of April 1, 1981 the Registry had 
comprehensive data on 864 patients who received 1,022 transplants performed 
rhy i6D -Xx-aqs plant teams. In addition, 78 follow-up reports were received m^' 
jiiixiag xixe past year. Each new and follow-up report was reviewed carefully 
and summarized by the Registry -staff . When necessary, transplant teams were 
contacted to obtain missing data for clarification of ambiquities in their 
reports. Computerization has resulted in an acceleration of the contractor's 
ability to perform analyses of the data. Detailed reports regarding 215 pa- 
tients with ALL, ANL, and CML who were treated with supralethal chemoradiother- 
apy and marrow transplantation between January 1, 1977 and December 31, 1981 
have been provided to the Registry by 28 worldwide bone marrow transplant 
teams . 



Significance to Cancer Research: While bone marrow transplantation has now 
become the treatment of choice for aplastic anemia and selected immune defi- 
ciency diseases and very promising results have recently been obtained in 
acute leukemia in remission, nevertheless a number of complications such as 
graf t-versus-host disease, recurrent leukemia, intercurrent infections and 
graft rejection remain major problems. The Registry provides the ability to 
look at the collective experience of many centers, which may facilitate more 
rapid progress in this form of treatment. 

Project Officer: Harriet L.G. Gordon, M.D. 

Program: Immunology Section 

FY 81 Funds: i 



100 



CONTRACT RESEARCH SUMMARY 
Title: Animal Models for Treatment of Minimal Residual Systemic Tumor 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-33891 
6/30/73 



Dr. Gerald L. Bartlett 
Pennsylvania State University 
Hershey, PA 



Expiration Date: 10/31/80 



Goal: To develop animal tumor models which will be useful for evaluating 
immune stimulants as immunotherapeutic agents. 

Approach: In four different tumor-host models, animals are given immunotherapy 
at various times after tumor implantation, either alone or in combination with 
other means of treatment. The effect of the immunotherapy and the role of 
immune stimulants in immunotherapy are evaluated in terms of improved sur- 
vival of treated animals. Animals which survive are tested further for im- 
proved tumor rejection immunity. 

Progress: 13762A Rat Mammary Adenocarcinoma . Rats cured by surgery after 
day 14 show strong tumor immunity. Concomitant immunity has been used to 
demonstrate the kinetics of tumor immnity in the treatment protocol employed 
and to detect immunity-stimulating treatments. Viable tumor cells (10°) 
are immunogenic, but irradiated tumor cells are not. In contrast, irradiated 
tumor cells can evoke DH but mitomycin C-treated cells cannot. Tumor immunity 
can be adoptively transferred by T-cells. 

Line 10 Guinea Pig Hepatoma . LlOX, which is an ineffective vaccine in 
male guinea pigs, is immunogenic for LlO in female guinea pigs that have been 
sensitized with a male skin graft. 

CaD2 Mammary Adenocarcinoma . Treatment with antithymocyte serum abolishes 
the therapeutic effect of i.t. Cp but not the effect of i.v. Cp. Drug induced 
tolerance to Cp has been induced. DH tolerance to Cp does not interfere with 
any of the tumor inhibitory properties of Cp in this model. 

LSTRA Mouse Leukemia . Chemoimmunotherapy (CY + vaccine) continues to be 
inconsistent, even in replicate experiments. Optimal adjuvants and optimal 
route of injection increase the efficacy of immunity by accelerating the 
induction of effective resistance to challenge tumor. LSTRA immunity is abol- 
ished by either prevaccine or postvaccine treatment with either ATS or CY, 
or by continuous treatment with trypan blue. 

Significance to Cancer Research: These studies demonstrate that caution must 
be exercised in drawing generalizations on immunotherapy from a single exper- 
raental or clinical result. 



Project Officer; Harriet L.G. Gordon, M.D, 
Program: Immunology Section 
FY 81 Funds: 



101 



CONTRACT RESEARCH SUMMARY W'' 

Title: Facility for Supplying Imniune-related Cell Lines 

Principal Investigator: Dr. Melvin Cohn 

Performing Organization: The Salk Institute 

City and State: San Diego, CA 

Contract Number: NOl-CB-23886 

Starting Date: 6/26/72 Expiration Date: 11/30/81 

Goal: To supply the scientific community with immune-related lines important 
in the study of tumor immunology and to increase the library of useful lines 
as they are characterized. 

Approach: Since this is essentially a contract to supply cell lines, little 
experimental work is involved. However, the general thrust of the laboratory 
involved the isolation of immune-related lines of the thymus and bone marrow- 
derived lineages by the use of leukemia viruses, cell fusions, and chemical 
carcinogens. As new lines become available, they are introduced into the 
contract catalog. 

Progress: The contractor has shipped 973 tissue culture and tumor lines to 
investigators in the scientific community over the past contract year. The 
catalog contains detailed characterization of the cell lines. This contract^ 
continues to be an excellent research resource increasingly utilized by " 
scientists all over the world. Computerization of inventory and shipments 
has greatly increased the efficency of the project. 



Significance to Cancer Research: The lines in this contract are used by 
hundreds of laboratories for studies on tumor-specific antigens, antibody 
structure, immune-related cell functions, somatic genetics, and cell fusions. 
All of these subjects are major categories under the NCI Cancer Research 
Program, and the cell lines from this laboratory are key tools In these in- 
vestigations. 

Project Officer: Ms. Judith M. Whalen 

Program: Immunology Section 

FY 81 Funds: $98,000 . 



102 



CONTRACT RESEARCH SUMMARY 

Title: Human Melanoma: Evaluation of BCG Immunotherapy of Patients Without 
Detectable Disease After Removal of Tumor-containing Lymph Nodes 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-64076 
8/8/74 



Dr. Donald L. Morton 
University of California 
Los Angeles, CA 



Expiration Date: 10/7/80 



Goal: To determine whether immunotherapy with BCG alone or BCG combined with 
tumor cell vaccine will decrease recurrence rate or prolong survival in mela- 
noma patients with metastases to regional lymph nodes. 

Approach: Patients are randomized following surgical lymphadenectomy into 3 
groups receiving different postoperative therapy: one group receives no addi- 
tional therapy, one receives BCG, and one group receives BCG and allogeneic 
tumor cell vaccine. 

Progress: Recurrences were least frequent in the vaccine treated group (23 
out of 47 - 49% compared to 27 out of 46-59% in the control group and 26 out 
of 45-57% in the BCG treated group). The differences between these recurrence 
frequencies is not, at present, statistically significant. Patients receiving 
BCG immunotherapy who recurred are still surviving longer than those in the 
control group, or those patients receiving BCG plus tumor cell vaccine. A 
blind analysis of stage II melanoma patients' sera for antibody against 
M14 did not confirm a previous observation that a modest rise in antibody 
levels following surgery correlated with a favorable prognosis. However, 
certain correlations could be made with the disease free interval and sur- 
vival when absolute levels of antibody were measured in the preoperative 
period. The mean titer of anti-Ml4 IgM antibodies was higher in the group of 
patients remaining tiomor free for two years relative to that in patients in 
whom disease recurred withn two years. A rise of complement fixation antibody 
titer greater than threefold in patient's receiving BCG with or without tumor 
cell vaccine was characteristic of patients whose disease did not recur with- 
in 30 months of entering the trial. Patients whose disease recurred with 6 to 
12 months did not show such a rise in CF antibody titer. 



Significance to Cancer Research: It is important to confirm in prospectively 
randomized trials earlier studies showing immunotherapy to be of benefit to 
cancer patients. 

Project Officer: Harriet L.G. Gordon, M.D. 
Program: Immunology Section 
FY 81 Funds: 



103 



CONTRACT RESEARCH SUMMARY 



Title: Studies of Immune Stimulants in Patients Receiving Radiation Therapy- 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-64004 
6/1/76 



Dr. William M. Wara 
University of California 
San Francisco, CA 



Expiration Date: 11/30/80 



Goal: To docximent the immunosuppression which occurs following radiation 
therapy and to determine if a therapeutic advantage can be achieved by the 
addition of immunotherapy (thymosin) to the conventional treatment given for 
squamous cell carcinoma of the head and neck region and the esophagus. 

Approach: Both groups of head and neck and esophageal cancer patients will 
receive radiation therapy and/or surgery as treatment. Patients will then be 
randomized to receive thymosin or placebo in addition to standard treatment. 
All patients will be monitored with immunologic tests to measure antibody- 
mediated and cell-mediated immunity at regular intervals. The data will be 
analyzed for local disease recurrence, metastatic disease, and survival 
difference in the patient groups. 



,t 



I 



ascertain quality control of the laboratory results. One hundred thirty sev 
patients have been studied and their preliminary results indicate the expected 1 
immunosuppression after irradiation. Following radiation therapy the adminis- I 
tration of thjnnosin did not appear to affect the total number of lymphocytes, 
total T lymphocytes, and B-lymphocytes. Head and neck cancer patients were 
normal pre-irradiation with respect to lymphocyte stimulation with PHA. 
Immediately, post irradiation the thymosin-treated patients showed less sup- 
pression than the patients not treated with thymosin, but the values for both 
groups were suppressed from the pre-irradiation levels. With respect to lym- 
phocyte stimulation by allogeneic cells (MLC), a significant difference is 
reported between the thjnnosin treated patients and those not receiving thjrmosin; 
the thymosin group retained their MLC capacity after immuno-suppressive irradia- 
tion. At three months post irradiation the differences in the PHA and MLC 
responses between the thymosin-treated and untreated patients became less 
apparent. 



Significance to Cancer Research: This study will determine the significance 
of immunosuppression caused by irradiation in head and neck and esophageal 
cancer patients and whether the effect of immunotherapy (thymosin) in rever- 
sing this immunosuppression has any impact on disease-free interval, survival, 
and local control of cancer. 



I 



I 



Project Officer: Harriet L.G, 
Program: Immunology Section 
FY 81 Funds: 



Gordon, M,D, 



104 



CONTRACT RESEARCH SUMMARY 
Title: Reagents for the Characterization of Human Cell Subpopulations 

Principal Investigator: Dr. Marius Teodorescu 

Performing Organization: University of Illinois 

City and State: Chicago, IL 

Contract Number: NOl-CB-84268 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal: Development of new reagents for the characterizaton of subpopulations 
of human cells important to the immune response or detection and character- 
ization of new subpopulations of these cells. 

Approach: Determine by _in vitro tests the function in immune response of 
lymphocyte subpopulations isolated by specific absorption of human lympho- 
cytes to a bacterial monolayer fixed on a glutaraldehyde-f ixed gelatin bed. 

Progress: The characterization of the T2 lymphocytes capable of suppressing 
natural killer cells has been completed. Previous work showed that the T1T2 
cells contain precursors for specific cytotoxic lymphocytes. Preliminary 
experiments demonstrate that the precursors of specifically cytotoxic lympho- 
cytes were exclusively located in Tl l3nnphocyte subpopulations. Moreover, 
after the activation of T cells in allogeneic mixed lymphocytes reaction, 
they maintained the same binding properties for bacteria. After activation, 
the cytotoxic T cells bind Bacillus globigii . This is particularly inter- 
esting since before activation, the cytotoxic lymphocytes do not bind these 
bacteria. Such results strongly suggest that non-activated specific cytotoxic 
cells only have the ability of binding Arizona hinshawii and Bacillus globigii . 
Therefore, it appears that the cytotoxic cells acquire a new receptor, probably 
a lectin that is identified by bacterial adherence. Additional experiments 
point out that the T3T4 cells are substantially more efficient in providing 
help in antibody formation. However, the results obtained so far, cannot 
establish exactly whether T3 or T4 cells are the actual helpers and additional 
experiments are underway to clarify these aspects. The identification of 
lymphocyte subpopulations using both bacteria and monoclonal antibodies sug- 
gest that the T1T2 cells are the same cells as those identified by 0K-T8 
monoclonal antibodies against T cell subsets producted by Ortho Laboratories 
and the T3T4 cells are the same as those identified by 0K-T4. Thus, it 
appears that the functions that we located in the T1T2 cells and in the T3T4 
cells are those shown by others to be performed by cells identified by the 
0K-T8 and 0K-T4 antibodies, respectively. It is noted, that bacteria have 
substantial advantages over monoclonal antibodies not only from the point of 
view of cost, but also from the point of view of accuracy, accessibility of 
morphology, simplicity etc. 



Project Officer: Ms. Judith M. Whalen 
Program: Immunology Section 
FY 81 Funds: 



105 



CONTRACT RESEARCH SUMMARY 



Title: Immunologic Mechanisms of Cattle 

Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Charles Muscoplat 
University of Minnesota 
St. Paul, MN 



Contract Number: 
Starting Date: 



NOl-CB-84244 
9/30/78 



Expiration Date: 9/29/81 



Goal: Perform in_ vitro studies of immunologic mechanisms in cattle in con- 
junction with ongoing immunoprophylaxis studies to gain information on the 
possible influence of immunity on carcinogenesis. 

Approach: Perform in vitro studies on the cellular and humoral events of the 
bovine immune system on tissues provided by NCI from experimental animals in- 
volved in a study of immunoprophylaxis of bovine ocular squamous cell carcinoma 
to determine whether correlative changes in immune response can be detected 
during the process of cancer development and, if so, to determine the immuno- 
logic mechanisms involved. 

Progress: A sensitive radioimmunoassay has been developed to detect anti- j 
bodies against cell surface antigens on bovine ocular squamous cell carcinoma ^ 
lines. Results show that most (if not all) animals with tumors possess anti-«^ 
bodies to their own tumor cells as well as allogeneic tumors. Differences in 
reactivity between autologous and allogeneic reactivity as yet cannot be 
determined. Normal cattle sera are non-reactive against all tumor cell lines 
as are tumorous cattle against normal corneal cell lines. These studies 
clearly indicate that it is easy to detect serum containing antibodies to 
tumor cells but difficult to distinguish one animal's tumor from another. 
Furthermore, in several instances the examination of plaques and papillomas 
which have originated from the same animal has not been able to distinguish 
them from one another using radioimmunoassay. Absorption experiments designed 
to determine cross reactivity of these antibodies show that all the tumor 
cells examined to date have a similar antigenic make up because all cells will 
absorb out antibodies to other cells (excluding normal controls). In studies 
involving natural cytotoxicity lymphocytes from normal and tumor bearing 
cattle have been examined to determine if lines derived from bovine ocular 
squamous cell carcinoma could be assayed for killing. Moderate levels of 
killing of several of these cell lines were observed. However, the degree of 
cytotoxicity varies with the passage of tumor cells in culture. It is not 
yet possible to detect differences in the lysis of tumor cells from different 
stages of tumor growth using natural cytotoxicity. 



Project Officer: Ms. Judith Whalen 
Program: Immunology Section 
FY 81 Funds: 



106 



CONTRACT RESEARCH SUMMARY 

Title: Reagents for the Characterization of Human Cell Subpopulations 

Principal Investigator: Dr. Noel Warner 

Performing Organization: University of New Mexico 

City and State: Albuquerque, NM 

Contract Number: NOl-CB-84288 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal: Development of new reagents for the characterization of subpopulations 
of human cells important to the immune response of detection and characteri- 
zation of new subpopulations of these cells. 

Approach: Develop specific heteroantisera against defined populations of 
human lymphoid cells stressing nonmalignant populations, including T-^, 1^ and 
T null cells, and other T cell preparations, as well as B cell populations 
defined by various available cell surface markers, and macrophage subpopu- 
lations . 

Progress: This study is intended to characterize the subset of human peri- 
pheral blood cells which bind to the Fc portion of human IgG (previously 
referred to T ^ cells) and to develop reagents that are exquisitely specific 
for these cells. Studies have demonstrated that this subset of cells expresses 
an Fc receptor that is capable of binding to monomeric human IgG of either 
myeloma or normal type, and thus by that criterion would be analogous to the 
Fc receptors so far identified only on macrophage lineage cells. Using a 
series of monoclonal antibodies from commercial sources it has also been 
demonstrated that these cells lack 0KT3 and other T cell specific antigens, 
but do express the OKMl antigens predominantly expressed on cells of the 
monocytic lineage. It is thus concluded that the majority of so-called 
T y cells are cells in the monocyte macrophage lineage. Using flow analysis 
the light scatter properties of these cells form in a relatively restricted 
medium scatter range distribution. Present studies are now in progress 
to develop monoclonal antibodies against these cells. These cells mediate 
natural killer activity on a range of human tumor target cells. These 
studies thus suggest that the monoclonal antibody raised against these cells 
might have the specific ability to inactivate natural killer- function or to 
monitor for NK function in a variety of human tumor target cells. In other 
studies a series of hybridoma fusions have been performed and are in varying 
stages of cloning, characterization and further development. 

Significance to Cancer Research: The development of reagents for characteri- 
zing subpopulations of human cells important to the immune response may 
facilitate analyses and manipulation of the immune response to human tumors. 

Project Officer: Ms. Judith M. Whalen 
Program: Immunology Section 
FY 81 Funds: 



107 



CONTRACT RESEARCH SUMMARY 
Title: Selective Depletion of Mononuclear Phagocytes In. Vivo 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-84271 
9/30/78 



Dr. Stephen Russell 
University of North Carolina 
Chapel Hill, NC 



Expiration Date: 5/1/81 



Goal: Development of methods for the selective in, vivo destruction or 
sustained functional inactivation of mononuclear phagocytes. 

Approach: Produce specific antimononuclear phagocyte globulin by selective 
absorption and assess the effect of these reagents on mononuclear phagocytes 
in vivo using "immune" granuloma formation in presensitized mice and spon- 
taneous regression of Moloney Sarcomas in mice as model systems. 

Progress: As was previously shown, the major cross-reactivity in the anti- 
bone marrow serum was to neutrophils. Therefore it was necessary to find 
the means to obtain large numbers of neutrophils free of mononuclear phago- 
cytes. Perironeal exudate cells collected 1, 2.5 or 4 hours after the in- 
traperitoneal injection of 1 ng LPS proved to be a rich source of neutro- 
phils. The neutrophils were found in the cell pellets with greater than m^ 
95% purity and only 1-3% contamination with mononuclear phagocytes. The 
contamination with mononuclear phagocytes was found to be caused by liquid | 
from upper layers mixing with the cell pellet. By pouring off the supernate 
and washing the sides of the inverted tube, the mononuclear phagocyte con- 
tamination was reduced to less than 0.5%. The number of neutrophils required 
to eliminate the anti-neutrophil activity of the anti-bone marrow IgG frac- 
tion was determined by titration. This laboratory has become increasingly 
aware of the potent effects of endotoxin on mononuclear phagocytes and, 
therefore, determined if endotoxin would affect their depletion assay. It 
was determined that 1000 ng/ml had a profound effect on the influx of mono- i] 
nuclear phagocytes into the peritoneal cavity after PHA stimulation. However,' 
even 50 ng/ml had a depressive effect on the influx of mononuclear phagocytes I 
when compared to PBS controls. 



Significance to Cancer Research: Studies on the _in vivo depletion of macro- 
phages will aid in examining the roles of macrophages in tumor destruction. 

Project Officer: Ms. Judith M. Whalen 
Program: Immunology Section 
FY 81 Funds: 



108 



CONTRACT RESEARCH SUMMARY 

Title: Immunoprophylaxis of Bovine Lymphosarcoma 

Principal Investigator: Dr. Richard M. Thorn 

Performing Organization: University of Pennsylvania 

City and State: Philadelphia, PA 

Contract Number: NOl-CB-84243 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal : Study the immunoprophylaxis of bovine lymphosarcoma through adminis- 
tration of BCG cell wall vaccine by intravenous injection as a possible guide 
for the control of analogous human disease. 

Approach: Study the effect of the administration of BCG cell wall vaccine 
on the extent and rate of development of lymphosarcoma in cattle injected 
with bovine leukemia virus at a stage prior to the development of discernible 
evidence of disease. 

Progress: One of the unusual immunologic phenomena associated with BLV- 
infected cattle is that their peripheral blood leukocytes spontaneously incor- 
porate thymidine in vitro . In this reporting period a systematic evaluation 
of the conditions for generating spontaneous blastogenic activity has been 
completed. The group demonstrated that only lymphocytes from BLV-infected 
cattle have a spontaneous blastogenesis which is inhibited by BLV antibody. 
Conditions have been standardized for the measurement of putative bovine 
T cells, which as judged by the literature, has been difficult. Of the 10 
cattle in this study which have died, 6 are known to have had lymphosarcoma. 
Since 3 were BCG treated and 3 were not, it appears that BCG did not have 
pronounced inhibitory effect. The number of cases is too small to evalulate 
the effect of BCG on the rate of deaths associated with lymphosarcoma. The 
2 cattle whose lymphosarcoma status at death is not yet known were BCG-treated. 
At present there is no evidence, therefore, that BCG-CWS given twice intra- 
venously reduces the incidence of naturally occurring, BLV-associated 
lymphosarcoma. Other studies imply that BLV infection induces a lymphocyte 
population which undergoes spontaneous blastogenesis in spontaneous DNA syn- 
thesis has been demonstrated in cultures of antigen specificity. This is, 
then, the first demonstration of specificity in a "spontaneous" immune re- 
sponse. This phenomenon may be unique to cattle and BLV infection or it may 
be a more general immunologic property which is easily detected in BLV-in- 
fected cattle because of the chronic nature of BLV infection. Plans are 
underway to determine the cellular basis and function of the inhibitable 
3STI activity found in lymphocytes from BLV-infected cattle. 



Project Officer: Ms. Judith Whalen 
Program: Immunology Section 
FY 81 Funds: 



109 



CONTRACT RESEARCH SUMMARY i 

Title: Immunoprophylaxis of Cancer Eye in Cattle 

Principal Investigator: Dr. Stephen J. Kleinschuster 

Performing Organiztion: Utah State University 

City and State: Logan, UT 

Contract Number: NOl-CB-74155 

Starting Date: 9/16/77 Expiration Date: 12/15/80 

Goal: To develop immunologic maneuvers for interfering with the progression 
of solid tumors from the premalignant (benign) to the malignant state. To 
relate the immunologic status of experimental animals to clinical events. 

Approach: Intralesional administration will be made of BCG cell wall vaccine 
into benign ocular lesions of cattle to determine whether such treatment will 
retard or prevent the normal progression of these lesions to the malignant 
state. Additionally, the immunologic status of animals will be monitored. 

Progress: The data shows, in general, that there were more regressed lesions 
in the active vaccine group than the others; however the significance level 
was variable. There were also fewer progressive lesions and malignant trans- 
formations in the active vaccine group than the others (high levels of signi-^ 
ficance). Thus, active vaccine administered intralesionally can interrupt tlH, 
transformation process and is a true prophylactic treatment. The collection 
and cryopreservation of tumor tissue and blood components was completed. 
Surgically removed tumor tissue has been frozen in saline (90° C). Bovine 
red cells are resistant to insult and can be frozen without elaborate prepar- 
ation. Bovine lymphocytes and monocytes, however, are much more fragile than 
their human counterparts. A protocol was developed for the successful cryo- 
preservation of bovine lymphocytes with a resultant 90% or greater viability 
upon thawing. Dr. C.C. Muscopiat has been supplied with all the blood com- 
ponents he will need for his analyses of immunocompetence of the experimental 
animals. Additionally, he has been supplied with approximately 35 cell lines 
from various animals in this study using techniques developed in the labora- 
tory. These lines are from all stages of the malignancy including normal, 
plaque, papilloma and frank carcinoma. 

Significance to Cancer Research: Studies involving the immunologic interrup- 
tion of the transformation of a benign precursor lesion to one of malignancy 
at the earliest possible stage provide valuable contributions to the overall 
cancer effort. Additionally, via a program of monitoring the immunocompetence 
of experimental animals as related to clinical events following experimenta- 
tion, relevant information can be contributed to the establishment of effec- 
tive immunologic testing needed in order to evaluate therapy and provide 
guidelines for improved human clinical protocols. 

Project Officer: Ms. Judith Whalen ^^ 

Program: Immunology Section 
FY 81 Funds: 



110 



I 



BREAST CANCER PROGRAM COORDINATING BRANCH 
October 1, 1980 - September 30, 1981 



Description 



The objectives of this extramural Program are to promote and support multi- 
disciplinary research projects in the laboratory and in the clinic that will 
lead to improved methods of diagnosis, prognosis, treatment and prevention 
of breast cancer. These objectives are accomplished through the collaborative 
activities of the. staff of the Branch and the members of the Breast Cancer 
Task Force Committee, an extramural advisory group. The members of the Task 
Force suggest new and innovative research ideas based on their knowledge of 
ongoing research in breast cancer as well as drawing upon their own expertise 
in the field; and information obtained at workshops and conferences in which 
the state-of-the-art on specific topics is presented. The staff of the 
Branch organizes the ideas into requests for investigator- initiated applica- 
tions (RFAs) or Program Announcements (PAs), or requests for contract 
proposals (RFPs) depending upon the mechanism of funding considered most 
advantageous to achieve the goals of the project. 

The Branch is organized into four program areas or Sections, namely. 
Diagnosis, Epidemiology, Experimental Biology and Therapeutics, with a fifth 
section on Information. The grants and contracts managed by each of the 
four Sections have been categorized by scientific relevance and are presented 
at the end of this summary. 

Accomplishments 

The grants program in breast cancer has increased in scope, in number, and 
in funding. In the previous fiscal year 46 grants for a total of approxi- 
mately $5 million dollars made up the program and in this year the number 
has increased to 88 for a total of almost $10 million. The extent of the 
present program is shown in tabular form. 

Both the Diagnosis and the Epidemiology areas have increased the scope of 
their programs with grants received in response to RFAs and Program 
Announcements. The Diagnosis area funded three grants from seven applica- 
tions received in response to a Program Announcement on research related to 
problems on mammographic screening. Each of the three projects initiated 
has a different approach to studying the effects of radiation on mammarv 
tissue or cells. The RFA in this area was related to studies of immuno- 
competent cells infiltrating human breast cancer, two of seven applications 
were funded. In the Epidemiology area the Program Announcement on genetic 
susceptibility to human breast cancer has been productive of applications, 
a total of nine, however, no awards have been made. The RFA on correlation 
between microscopic characteristics of primary breast tumors and subsequent 
patient survival resulted in receipt of 17 applications. Four of these 
received excellent scores however the funds set aside for the RFA permitted 

dijL'^%""'''^''°'? °^ ^"° g^^^t^- ^11 f°- applications represented 
different scientific approaches to the problem therefore it would have b 



een 



111 



5reast Cancer Research Program 
Fiscal Year 1981 



Project 






Grants 




Contracts 


Category 


Number 






Funds 


Number 


Funds 


Experimental Biology 




48 




$4. 


,859,841 


10 


$ 148,500 


Epidemiology 




11 




2., 


,003,258 


14 


274,000' 


Diagnosis 




14 




1; 


,441,921 


10 


27,000 


Treatment 




15 




1., 


,593,646 


17 


783,918^ 


Totals 


88 


$9. 


,898,666 


51^^ 


$1,233,418 


Grand Totals - Grants 


and 


Contracts 




139 


$11,132, 


,084 



a $80,000 Provided by the Nutrition Program, NCI 

b $105,073 Provided by the Office of the Director, NIH 

c 19 Contracts Received Funds 



112 



I 



highly desirable to have funded each. The applications to the two Program 
Announcements were reviewed by regular Study Sections while those to the 
RFAs were reviewed by Special Study Sections. 

Although the Experimental Biology and Treatment areas have not advertised 
new projects each has increased their program. Some of the new grants 
funded in the four areas can be attributed to the encouragement of investi- 
gators by staff of the respective Sections to apply for support for new and 
relevant breast cancer projects. Both in the areas of Experimental Biology 
and Diagnosis, research related to the production and characterization of 
monoclonal antibodies for breast cancer has received initial support. 

The grants of the total program include: 79 ROls-Regular Grants, competitive 
and renewals; 2 POls-Program Project Grants, one basic research, one clinical; 
5 R23s-New Investigator Awards; and, 2 R23s-Conf erence Grants. 

Projects funded by contracts have continued to decline, in the previous 
fiscal year there were 80 funded for a total of $3.1 million; this year 
(FY 1981), there were 51 however only 19 received funds, a total of $1.2 
million. Those not funded but continuing were either completing their final 
year or received extensions without additional dollars, the majority of the 
remaining contracts are follow-up of clinical studies. The two contracts 
that received the most dollars are service in nature. 

The Task Force Committee membership has been reduced from 38 to 27 members 
in Fiscal Year 1981. On several occasions it has been necessary to invite 
one or more ad hoc investigators for a meeting in order to have expertise 
that was lacking within the present membership. The Committee has met four 
times during the year and at each of the first three meetings one half-day 
was devoted to a workshop or scientific session, at the last meeting a full 
day workshop was held. Topics for the workshops have been suggested by 
members of the Committee and in some instances members have presented their 
own work. The topics and sponsoring Program areas were: 1. "Luteal Phase 
Defects and Breast Cancer Risk" - Epidemiology. 2. "Clonogenic Assays and 
Chemotherapy Sensitivity" - Treatment. 3. "Chemical Carcinogen-Hormone 
Interaction in Transformation of Mammary Epithelial Cells rn Vitro " - 
Biology. 4. "Monoclonal Antibodies in Breast Cancer" - Diagnosis. Addi- 
tionally the Epidemiology Program area in collaboration with the Diet, 
Nutrition and Cancer Program, NCI, sponsored a day working session on "Diet 
and Breast Cancer Risk;" and, it was also responsible for bringing three 
speakers to present their work on "Specific Dietary Factors in Relation to 
Benign Breast Disease or Cancer." The specific topics were: abstenance 
from intake of methylxanthines , coffee, tea, colas, and chocolates; Vitamin E 
(tocopherol) supplement to diet; and a literature review on studies of 
Vitamin A and cancer. Attendance at these meetings has been excellent, 
Task Force members, NCI and other NIH staff, basic and clinical investigators 
from universities and other research organizations in this country and 
abroad, pharmaceutical organizations and lay persons have attended. Only 
Task Force members and invited speakers have been reimbursed for participa- 
tion. The purpose of a workshop is to review the state-of-the-art and to 
determine the need for promoting new research. Members of the staff of the 
Branch have been preparing projects to be advertised for RFAs, Program 

113 



Announcements and RFPs resulting from suggestions made at the time of each^ 
workshop. All projects must be approved by the DCBD Steering Committee and 
the DCBD Board of Scientific Counselors before they are published. Because 
of the time of the meetings of the Board no new projects have been published 
in this fiscal year. 

In the coming year the following research projects and others will receive 
emphasis from the staff and advisors: monoclonal antibodies, diagnosis and 
treatment; new diagnostic methods, particularly diaphanography and NMR; 
improved techniques for clonogenic assays; specific dietary factors in 
relation to benign breast disease and cancer; genetic markers in high risk 
families; identification of intravascular tumor growth in histological 
sections; and, luteal phase defects and breast cancer risk. 

Diagnosis Section 

The Section continues to be concerned with major clinical issues that con- 
front the physician in the diagnosis and prognosis of breast cancer. These 
issues encompass several medical disciplines and include such problems as 
early diagnosis, separation of non-neoplastic from early neoplastic condi- 
tions, staging, determinants of biological behavior and predicting prognosis 
and recurrence. The section has supported through grants and contracts basic 
and applied research related to these issues. Although some may feel that 
progress is slow, we confidently believe that definite advances have been 
made. Studies on breast cancer are complicated by the nature of the lesio 
and by the many endocrine and other factors that affect breast tissue. Th 
following describes different projects which we have developed to address 
these issues: 



\ 



:y 



Biological Markers : 

Efforts to identify biological markers continue to receive special attention. 
The importance of markers lies in their many potential uses. For instance, 
they might detect incipient cancer, confirm a clinical diagnosis, or stage 
a patient for therapy. They may prove useful in estimating prognosis and 
predicting recurrences. Currently, the three most useful markers are 
ferritin, calcitonin and CEA. Unfortunately, these are not reliable for 
early detection and cannot always be used to predict prognosis. The Section 
has awarded one grant and six contracts in its search for biological markers. 
One of our objectives in marker research (NOl-CB-84222) is to quantitate 
and characterize the different isozyme patterns in normal, benign and 
neoplastic breast tissue. Although the isozyme patterns of normal and 
malignant tissues are known to differ, we would like to take advantage of 
this difference to find serum or tissue enzymes that reflect the presence of 
cancer. Of 23 different isozymes tested only three seem promising: lactic 
dehydrogenase, malic acid of dehydrogenase and esterase. Because these 
isozymes exist in tissues, they can be detected by histochemical methods, 
which might prove helpful to the pathologist for differentiating borderline 
lesions . 

In another study (CA25574) the isozyme differences and the effects of 
mammotropic hormones on these differences are being investigated. 

114 



Concerning prognosis, the Section has supported longitudinal studies 
(CB-74086 and C-B-74206) in breast cancer patients to learn if long-term 
changes in the levels of serum enzymes or other constituents can be used to 
predict recurrence. Although time consuming, these types of studies are 
especially valuable. Based on information from the initial presurgical and 
surgical blood data, a complex multi-variate model has been developed that 
will now be tested in a prospective study. Based on the recurrent patterns 
of patients followed for a period of time, this model successfully predicted 
nearly 80% of recurrences. It should be noted, however, that the relation- 
ship of putative serum markers to recurrence is not always clear. For 
instance, in some patients with metastatic breast cancer, elevation in 
markers preceded the clinical metastasis while in other patients no changes 
occurred. In addition to correlating changes in the serum levels with time, 
investigators are also exploring the relationship between marker concentra- 
tion in tumor tissue and its level in the blood. 

In addition to malignant diseases, we have turned to benign breast diseases 
in an effort to identify new markers, especially those that may correlate 
with the subsequent development of cancer. Contract CB-53853 has analyzed 
fluid aspirated from benign mammary cysts. Results from over 1600 samples 
showed that CEA, alpha-fetoprotein, the beta subunit of human chorionic 
gonadotropin, calcium and other metals, cholesterol, numerous enzymes, 
steroids and cell breakdwon products are present in the cystic fluid in much 
higher concentration than in serum. These patients are now being followed 
to establish whether the development of cancer, which is expected to occur 
in a certain number, will correlate with the initial biochemical findings. 
These types of studies are extremely time consuming, but they are considered 
very useful. 

Other research projects (CB-84223 and CB-84316) have been concerned with 
distinguishing premalignant hyperplasia from non-premalignant hyperplasia. 
These studies involved the extensive use of biochemical, morphologic and 
immunologic techniques in order to associate putative markers in tissues 
with malignancy. They should have practical significance if specific 
markers can be identified that are easily measured on routine histologic 
sections. Results from these projects have shown that CEA is present in 
malignant lesions and in severe epithelial dysplasia, but only rarely in 
cases of fibrocystic disease. IgG has been found in a pericellular pattern 
in dysplasia while IgA predominates in non-dysplastic benign lesions. The 
presence of IgG seems to correlate with lymphopenia. The full significance 
of these remains to be established. Angiogenic factor, which stimulates 
vascularization, has been found in a large number of atypical breast lobules 
obtained from women with carcinoma and in a smaller number of atypical 
lobules obtained from women without cancer. 

Methods of Detection : 

Regarding non-invasive methods of detection, two grants have been initiated. 
One (CA25836) is designed to test the feasibility of localizing breast 
lesions by gamma imaging techniques using radiolabeled steroidal and other 
derivatives that bind specifically to estrogen receptors. A number of gamma 



115 



labeled iodostilbestrol and halogenated hexestrols that bind to these ^ 
receptors have been synthesized and characterized. These compounds are 
presently undergoing ±n vitro and ±n vivo testing. 

Studies on diaphanography and nuclear magnetic resonance have been investi- 
gated and will receive more attention in the coming year. 

One study (CA28967-01) was recently funded in which a scanning electron 
microscope is used to magnify the images of microcalcif ication obtained by 
low dose mammography. The purpose of this investigation is to determine if 
early granular calcification indicates the presence of cancer even before 
its pattern can be discerned on routine film examination by the radiologist. 
Although it is impractical to use a scanning electron microscope routinely, 
the results of this study should tell us if the pattern of microcalcif ication 
is established early in the evolution of neoplasia and if it can be used for 
early diagnosis. 

Another long-term concern has been adverse effects of diagnostic modalities. 
For instance, three grants (CA29940, CA29993, CA29781) in response to a 
Program Announcement were recently awarded to study the effects of irradia- 
tion on mammary carcinogenesis in rats and DNA damage repair in mammary 
cells. 

Morphologic Discriminants : 

Accurate diagnosis and morphologic assessment of breast cancer remains a ^ 
challenging problem for the" Section. The statistics regarding the incidenS: 
of breast cancer indicate the need for early and effective diagnostic 
evaluation. Interest in early evaluation includes an assessment of the 
probability of invasive cancer following hyperplasia or in situ cancer as 
well as in histologic markers for the separation of borderline lesions. 

In order to ascertain the risk of invasive cancer with in situ and dysplastic 
lesions, we have been supporting a histologic review (CB-74098) of a large 
number of carefully followed women with an initial diagnosis of ^ situ 
carcinoma, atypical hyperplasia or other benign conditions. This work is of 
practical importance because it may improve diagnostic criteria pathologists 
use to estimate the biologic behavior of these lesions. The histologic 
evaluation of dysplastic and other atypical lesions has been controversial 
for years. 

Studies to define whether expression of common blood group antigens, such 
as the ABO system and T-antigen, on the primary cancer can be used to 
predict metastatic spread (CA28128) . It is also possible that these blood 
group antigens may also be useful in differentiating hyperplasia from early 
malignancy. Results with the T-antigen indicate that the pattern of intra- 
cellular distribution may be important. T-antigen was found only along the 
apical cytoplasmic membrane in 19 of 22 benign breast lesions whereas a 
diffuse cytoplasmic distribution was seen in 17 of 22 cases of breast cancer. 
The carcinomas which did not display the T-antigen were the most poorly 
differentiated of the group. ^ 



116 



A marker (CA26406) has been found that differentiates mannnary epithelial 
cells from mesenchirmal cells and from myoepithelial cells. This marker 
should separate lesions originating from ductal or lobular cells as opposed 
to those arising from myoepithelium. The marker is actually keratin, which 
occurs in small amounts in epithelium. 

Immunodiagnosis : 

The potential clinical use of immunological methods for diagnosis and for 
estimating prognosis is a critical concern. There are four grants and two 
contracts relating to immunodiagnosis in breast cancer. Although in the 
past studies were usually directed at correlating prognosis with the extent 
of the inflammatory response to the tumor, more recent studies have been 
toward measuring the functional ability of these lymphocytes. For example, 
contract CB-84228, has been assessing the prognostic significance of 
the cell-mediated immune reactivity in standarized in vitro tests. The 
results firmly suggest that the tumor does affect functional activity in 
regional l3^mph nodes, an observation that may have bearing on the metastatic 
potential of breast cancer. This field should continue to attract con- 
siderable research interest for there is little information concerning the 
function of intratumor leukocytes and their relation to any immune response. 
In this regard, we have recently awarded two grants (CA29601, CA29586) to 
investigate immunocompetent cells that infiltrate human breast cancer. 

In addition, research designed to correlate the level of circulating antigen- 
antibody complexes with prognosis, estrogen receptor levels and other 
variables (CB-84224 and CA25653) has been supported. Circulating immune 
complexes are found in many patients with cancer including those with breast 
cancer. Following an analysis of over 400 patients, immune complexes 
measured by several different techniques have been found in nearly all breast 
cancer patients and in some patients with benign breast disease. As 
expected, the levels of these immune complexes have varied with the differ- 
ent methods used. According to preliminary analysis, the level correlates 
with the estrogen receptor content of the tumor and certain risk factors. 
Patients with Stage 2 have higher levels than patients with Stage 1 disease. 
Levels do not correlate with circulating CEA levels. The results also 
indicate that the Raji assay is most effective in differentiating cancer 
from non-cancer groups. 

The levels of immune complexes vary depending on past history, especially 
in patient with a history of genital urinary tract infection or hyper- 
tension. These latter observations are not only important for breast 
cancer patients, but for all cancer patients. One study that remains to be 
done is to determine the nature of the antigen that forms the immune 
complex. In addition to evaluating the level of immune complexes in breast 
cancer patients, the investigators are improving the techniques for measur- 
ing these complexes. 

Also skin tests are being evaluated as a possible diagnostic tool (CA19083) . 
There is evidence that the Thomasen Frendenreich (T) antigen which is a 
carbohydrate precursor antigen for the MN blood group system is specifically 
associated with adenocarcinomas of the breast. Preliminary data indicate 



117 



that breast cancer patients and a small number of patients with benign ^ 
disease show both humoral and cell-mediated immune reaction to the 
T-antigen. As part of a prospective study, these antigens are being 
measured in patients without known breast cancer and in patients who will 
have a breast biopsy for suspected malignancy. 

Other Activities : 

In collaboration with the College of American Pathologists, the Section 
organized a symposium on "The Management Assessment of the Aspiration 
Needle Biopsy in Breast Cancer," which was held in San Diego during the 
spring meeting of the College. 

Workshop on "Monoclonal Antibodies in Breast Cancer." 

Epidemiology Section 

Fourteen contracts and eleven grants are categorized under nine different 
areas of focus in our epidemiologic studies of breast cancer. Five of 
these represent more than one category, since the categories are intertwined; 
one of these five (CA13556) is a program project grant with a broad, mulit- 
disciplinary approach to the etiology, epidemiology, and natural history 
of breast cancer. Seven of the fourteen contracts (CB-53884, CB-53968, 
CB-63996, CB-63997, CB-74102, CB-74103, CB-74104) terminated in Fiscal Year 
1981; one of the other seven ongoing contracts was extended without funds. 
Eight new grants have been added to the program this year; two of these u 
were top-scoring responses to a specific RFA. 

In the area of genetic susceptibility to breast cancer, one grant (CA27632) , 
following up our previous contract-funded research by this same investigator, 
is extending research stenmiing from her very important discovery that breast 
cancer susceptibility appears to be linked, in some but not in all breast 
cancer prone families, to the locus for glutamine pyruvate transaminase 
(GRP-1) , a marker gene provisionally located on chromosome 10. This strong 
evidence for at least one gene for breast cancer susceptibility is a major 
breakthrough that has opened up an entire new realm of research, including 
their continuing attempt to localize other genes for susceptibility through 
similar linkage analysis, as well as the search for the phenotypic expres- 
sion of such a gene. A related grant (CA30069) is exploring hormone 
metabolism patterns associated with familial high risk. 

The endocrinology of breast cancer etiology has continued to be explored 
through a variety of studies: A. Examining exogenous estrogens as a risk 
factor (CB-53884, CB-74099). B. Seeking differences between pre- and 
postmenopausal breast cancer (CB-63996). C. Defining endocrine events at 
the time of first pregnancy in young and in older women (CB-74101) . 
D. Exploring epidemiologic risk factors in relation to steroid hormone 
receptors and hormone binding globulin (CA13556, CB-63996), and examining 
these in relation to prognosis (CA28720). E. Studying hormone metabolism 
associated with familial breast cancer (CA30069). F. Searching for possible 
relationships between breast cancer and prior thyroid diseases (CB-84230) . ^ 



118 



Interaction with pathology has involved explorations of possible differences 
in_ the epidemiology of different histopathological subcategories of breast 
cancer (CB-53968, CB-63997) . 

Program interest in the natural history of breast cancer has led to studies 
on the epidemiology of benign breast disease(s), (CB-74102, CB-74202, 
(CB-84231, CA13556, CA26021, CA28720) and of minimal breast cancer 
(CB-74103). A particular focus is the epidemiology of benign lesions most 
prone to progress to breast cancer. Incidence of benign breast disease is 
also being examined in ethnic subsets of the population known to differ in 
their risk of breast cancer (CB-84231). In relation to natural history, a 
mathematical two-hit model was developed for the pathogenesis of breast 
cancer; this model was in good agreement with available incidence and 
epidemiologic data (CB-74100, completed in Fiscal Year 1980). 

Concern about environmental risk factors has continued; attention has been 
focused not only on exogenous steroid hormones, but also on other possible 
carcinogens, and particularly on the possible role of diet (CB-53884, 
CB-74104, CB-84229, CB-84318; CA30273, CA30629). Special emphasis on 
cholesterol and lipids has been timely in terms of present interest in a 
possible negative correlation between risk for cardiovascular disease and 
risk for cancer. The principal finding in terminated contracts CB-84238 
and CB-84317 (terminated in Fiscal Year 1980) was that there was no relation- 
ship between prior serum cholesterol level and the subsequent risk of 
development of breast cancer. Another ongoing contract (CB-84318) is 
continuing to explore differences between breast cancer cases and controls. 
Emphasis has been placed on possible mechanisms and rationale for a dietary 
effect. The effect of obesity and of weight loss on steroid hormone 
metabolism is being determined (CB-84229). Significant differences have 
been found between vegetarians and omnivores in the excretion of estrogens 
and the metabolism of steroids by the intestinal flora (CB-74104). Dietary 
fat is being examined in relation to physical chemical properties of the 
cell membrane, immune responsiveness, and tumor cell susceptibility to 
cytotoxicity (CA30273, CA30629) . Carcinogens and mutagens that may partic- 
ularly impinge on breast ductal epithelium are being determined from 
examination of breast fluids (CA13556). 

Interest has also focussed on risk factors of possible relevance to tumor 
agressiveness and hence to prognosis; new stuides are aimed at exploring 
specifics of epidemiology and of the morphology, biochemistry, immunology, 
and immunohistochemistry of breast tumors in relation to prognosis and 
survival (CA28720, CA30335, CA30342). These will include attention to 
antiviral antigens in relation to breast cancer risk and to prognosis of 
breast cancer (CA29711, CA30342). 

The overall emphasis of the Epidemiology Program is not simply statistical, 
descriptive epidemiology, but rather a broad, multidisciplinary approach 
to the exploration of possible etiologic mechanisms and the risk factors 
influencing these. Program has maintained a continuing relationship with 
the Diet, Nutrition, and Cancer Program (DNCP). The Section Chief serves 
as a member of the NCI Nutrition Working Group, and represents the Division 
of Cancer Biology and Diagnosis on that group. In this capacity, she has 



119 



participated in the discussions and decisions on policies and reconmienda- ^ 
tions regarding NCI funded nutrition research, and in their interaction ▼ 
with the Nutrition Subcommittee of the National Cancer Advisory Board. As 
mentioned early in this report the Epidemiology Section and DNCP cosponsored 
a Working Session on "Diet in Relation to Breast Cancer Risk" in April 1981 
that attracted a large attendance and valuable discussion on future avenues 
of research in the area. 

Experimental Biology Section 

The program maintains the largest number of investigator- initiated research 
projects that encompass a variety of scientific approaches to basic biology. 
Five contracts were terminated in Fiscal Year 1981 (CB-74094, CB-74188, 
CB-8A134, CB-84225 and CB-84239) and contract (CB-74175) continues to serve 
as an animal tumor and human breast cancer cell culture bank. Contracts 
(CB-84134 and CB-84225) are service oriented for the production of anti- 
bodies to various types of collagen and the carboxyterminal non-helical 
sequences of human Type I procollagen. Contracts (CB-74094 and CB-74188) 
are research oriented to determine factors for growth and passage of normal 
mammary epithelial cells in culture. The largest number of grants were 
assigned to the Section resulting in a larger number of funded grants. 
There were 48 grants in the program in Fiscal Year 1981 that are competitive 
and non-competitive, one Program Project, three New Investigator Awards and 
one Conference Grant. 

The funded grants conduct research relating to various categories such as : _. 
I) Factors in the Inductidn of Mammary Tumors (CA18946, CA27026, CA27293, % 
CA20764, CA18664, CA25915, CA31207, CA21993, CA22879, CA28999, CA30036, 
CA31774 and CA30570) ; II) Growth Passage and Characterization of Normal 
Mammary Epithelial Cells (CA05388, CA24844, CA29090, CB-74094 and CB-74188); 
III) Mammary Tumor Response to and Dependence on Hormones (CA16660, CA21606, 
CA20605, CA30171, CA26869, CA18664, CA27293, CA28645, CA28698, CA28393, 
CA18458, CA16464, CA22343, CA24687 and CA29501) ; IV) Interaction of Neo- 
plastic Mammary Cells With Other Cell Populations (CA25179, CA20764, 
(CA20122, CA28366 and CB-84239); V) Cell Surface Proteins in Relation to 
Metastasis (CA19814, CA27909, CA31695, CA19455, CA08418, CA27314, CA26825, 
CA25418 and CA28844) ; VI) Prevention of Neoplastic Transformation (CA30036, 
CB-74207) ; VII) Pathophysiology of the Solid Mammary Carcinoma and its 
Relation to Metastasis (CA05388, CA29537 and CA28735) ; VIII) Immunodiagnosis 
and Immunoprevention of Cancer Growth and Progression (CA27437, CA30284 and 
CA20286); IX) Service, Support and Resource (CB-74175, CB-84225, CB-84314, 
CA28816 and CA30294) . Because of the types of research grants some were 
included in two or more categories as CA05388 in categories II and VII; 
CA18644 and CA27293 in categories I and III; CA20764 in categories I and 
IV; CA30036 in categories I and VI. A research summary on each contract 
that includes information on goals, approaches and progress, as well as, a 
list of the grantees' institutions, principal investigator, grant numbers 
and titles of the research projects appends this report. 

A Workshop entitled "Chemical Carcinogen-Hormone Interaction in Transforma- 
tion of Mammary Epithelial Cells In Vitro " was sponsored in April 1981. 
Six investigators with expertise in this scientific area were invited to ^ ) 



120 



present their latest research. The workshop was very successful and the 
concensus was that epithelial cell culture techniques and research methods 
are available to study the epithelial transformation process which hitherto 
was done with fibroblasts. 

Therapeutic Section 

This program is primarily concerned with developing new knowledge about 
tumor biology which can be applied to the optimal selection and utilization 
of various treatment modalities. 

Fifteen contracts and fifteen grants are listed under five major research 
areas. In addition, the data management center constitutes a resource 
contract (CB-14339) . Five contracts were terminated in Fiscal Year 1981 
(CB-33899, CB-43917, CB-74137, CB-74140 and CB-84221) and four were confined 
to clinical follow-up observations. 

For more than a decade the program has been deeply involved with steroid 
receptor technology and interpretation as a means of predicting hormone 
dependency of breast cancer tissue. Three grant supported studies 
(CA22828, CA26452 and CA27470) are directed toward elucidating the physio- 
logic activity of the steroid receptors. With the trend toward receptor 
assays being performed on all patients with primary and metastatic breast 
cancer, there is need for procedures that are simpler and more reproducible 
than the standard methods. There also is need for methods that will 
demonstrate the histologic distribution of the receptor proteins. One 
grant supported project (CA29971) and two contracts (CB-23862 and CB-43969) 
have these objectives. An evaluation-oriented contract (CB-04388) funded 
from the Office of the Director, NIH, is sampling the impact on clinical 
management of breast cancer produced by the application of steroid receptor 
data. 

The search for biological markers in breast cancer patients continues. 
Four contracts (CB-74137, CB-74138, CB-74213 and CB-84296 are resource 
oriented with the aim of establishing a bank of serum specimens from which 
qualified investigators may draw panels of coded specimens to test their 
purported markers. Three medical centers are processing serum samples from 
1) asymptomatic women, 2) women with benign breast disease and 3) patients 
with breast cancer. The samples are stored in a Mayo Foundation facility 
where they are coded and held for distribution to the investigators. 

Research studies in the biomarker area are being carried out by one grantee 
and two contractors (CA26935, CB-43900 and CB-74204) . Dr. Tormey is testing 
CEA and FHAP as breast cancer management parimeters. Dr. Dao is studying 
sulfotransferase enzymes as clinical prognostic markers and Dr. Hilf is 
measuring a panel of tumor isoenzymes as potential prognosticators of 
response to chemotherapy. 

A group of seven grant supported projects and one contract (CA020 71, 
CA05197, CA23079, CA24129, CA25586, CA26004, CA30251 and CB-84221) deals 
with the mechanisms by which chemical and endocrine treatments cause 
alterations in breast tumor growth. They explore specific tumor, host or 



121 



treatment factors such as cytokinetics, innnuno logical parameters, endocrin« 
profiles during cytotoxic therapy, and intratumor steroid metabolism. " 

Animal models for preclinical treatment testing are the major thrust for 
two grant studies (CA26287 and CA29006) ; a rat mammary carcinoma system for 
c_. parvum and canine breast cancers for envelope glycoprotein 55. 

The final category is clinical investigation with five contracts and one 
grant (CB-33899, CB-43917, CB-43990, CB-53851, CB-53917 and CA30006). Four 
of the contract-supported studies are involved with systemic therapy, 
adjuvant to local therapy, for patients with tumor localized to the breast 
and axillary nodes. The fifth tests endocrine suppressive therapy against 
advanced breast cancer. The adjuvant studies have completed patient accrual 
and the courses of treatment and are in the long-term follow-up phase. The 
advanced breast cancer project has also finished patient enrollment but 
continues with in depth endocrine studies. One of the adjuvant studies 
showed an advantage for patients with ER-positive cancers who received the 
anti-estrogen Tamoxifen in addition to CMF . The investigators have 
developed further treatment protocols and the new project will be grant 
supported (CA30006) . 

The Workshop on "Clonogenic Assays and Chemotherapy Sensitivity" was held 
in February 1981. A staff activity was the issuance of RFP NCI-CB-14339: 
Biomedical Computing Software Services in Support of Breast Cancer Treatment 
Program. A number of excellent proposals were submitted and the transition 
to the new contractor has gone smoothly. The Data Center is heavily involupd 
with the Biomarkers project, classifying all specimens, selecting panels ^ 
for distribution and analyzing the test results. 

Information Section 

The responsibilities include: 1) The monthly production of the publication 
Intercom which provides up-to-date listings of scientific papers on breast 
cancer research in biology, epidemiology, diagnosis, and treatment by title, 
author and journal reference; a list of meetings and conferences related to 
the disease; and, abstracts of presentations made at Workshops. The publi- 
cation is sent to investigators and institutions throughout the world, 
approximately 1700 copies are distributed monthly. 2) A monthly publication 
of Intracom which is similar to the listing of the scientific literature 
found in Intercom , however, it provides abstracts or summaries of the 
published literature as well as those on grant and contract activities. 
The publication is provided only to members of the staff of the Branch and 
to the Chairman of the BCTF Committee. 3) Responding to inquiries for 
literature searches by subject matter and by investigator. The resulting 
information is provided to staff, both of the Branch and to others in NCI, 
and to members of the Committee as well as to grantees or contractors. 

The principal developments of Fiscal Year 1981 have been: 

INTRACOM software has been enhanced to incorporate various improvements 
which include an author index and production on microfiche, Distributi^, 
of the microfiche publication to the Task Force Committee members has V' 



122 



begun. Opinion solicited by questionnaire has disclosed that some lack 
enthusiasm for the microfiche medium, although substantially all have 
access to a reader. About 1/3 of the group responded and this ratio, 
applied to the INTERCOM list, suggests that there are about 500 potential 
users world-wide. Under this assumption, global distribution can be 
managed with available resources at an annual cost of $7,500-10,000 for 
all charges, including postage and handling. The annual incremental 
cost for an additional user is $12-15. 

PROGRAMMING has begun for the fast search service to be made available 
to the Committee members and ultimately to others. This service will 
transmit output by telecommunication and provide a turnaround time of 
1-2 hours for many searches unless overload occurs. Thereby the breast 
cancer workers will be freed from the long delays which attend search 
queries submitted through library centers and will be able to exploit 
more effectively the growing power of automated information systems. 
The communication interfaces required have been installed on the 990 
minicomputer. 

SEARCH REQUESTS from the staff continue to increase, covering a wide 
range of subjects and sophistication. Capacity is ample, although 
turnaround time suffers occasionally when other work is pressing. 

ADP UTILIZATION has not been fully effective in augmenting clerical 
productivity but further improvement should be possible with the major 
enchancement of WYLBUR introduced recently by DCRT. 

Future Course 

INTRACOM will receive a subject index as soon as the impending addition 
of medical subject headings to the CANCERLIT file is done by MEDLARS. 
The subject index will require a substantial programming effort but will 
complete the present plan for the development of INTRACOM. 

The distribution of INTRACOM will be expanded. 

SEARCH SERVICE by telecommunication will be instituted experimentally 
and evaluated for utility. 

TELECONFERENCING, mentioned for the future in the previous report, was 
not reached but programming for it and some experimenting with it will 
be done if there is sufficient interest. This project will not be 
started until experience with the search service has provided some 
information concerning the terminal facilities of the users. 

TRAINING of clerical personnel in the use of ADP procedures will continue. 

The significance of the program is the facilitation of access to comprehensive 
and timely information concerning breast cancer that should enhance the 
quality of program formulation by staff and consultants. 



123 



124 



BREAST CANCER TASK FORCE GRANT SUPPORTED STUDIES AND ONGOING CONTRACTS 



DIAGNOSIS 

I. Biophysical Tools for Detection and Diagnosis 

ROl-CA-25836 

Rational Design of Breast Tumor Localizing Agents 
Katzenellenbogen, John University of Illinois 

ROl-CA-28961 

No-Dose Highly Magnified Mammograms to Aid Diagnosis 
Galkin, Benjamin Thomas Jefferson University 

ROl-CA-29993 

Effects of X-Rays on Human Mammary Epithelial Cells 
Smith, Helena University of .California, Berkeley 

ROl-CA-29940 

Damage-Repair Studies Related to Mammography 

Han, Antun/Elkind, Mortimer Argonne National Laboratory 

ROl-CA-29781 

Effects of Dose Rate on Rat Mammary Carcinogenesis 

Shellabarger, Claire Brookhaven National Laboratory 



II. Immunodiagnosis 

ROl-CA-25653 

Antigen-Antibody Complexes in Breast Cancer 
Chu, Tsann Roswell Park Memorial Institute 

ROl-CA-19083 

T-Antigen in Human Cancer Detection 
Springer, Georg Evans ton Hospital 

ROl-CA-29586 

Immunocompetent Cells Infiltrating in Human Breast Cancer 
Carmack, Holmes University of California, Los Angeles 

ROl-CA-29601 

Immunocompetent Cells Infiltrating Human Breast Cancer 
Bhan, Atul Harvard Medical School 

R23-CA-30370 

Monoclonal Antibodies to Breast Cancer Immune Complexes 
Papsidero, Lawrence Roswell Park Memorial Institute 



125 



'''NOl-CB-84228 

Cunningham-Rundles, Susanna Sloan-Kettering 

NOl-CB-84224 

Medof, Edward University of Chicago 



III, Biologic Markers in Breast Cancer Preneoplasia 

ROl-CA-25574 

Isozymes Specific to Human Breast Neoplasia 
Yang, Ning-Sun Michigan Cancer Foundation 

ROl-CA-30636 

Human Immune Responses to Murine Mammary Tumor Virus 
Dion, Arnold Institute for Medical Research 

NOl-CB-84222 

Balinsky, Doris Iowa State University 

NOl-CB-84316 

Jensen, Hanne University of California, Berkeley 

NOl-CB-84223 

McCarty, Kenneth, Jr. Duke University 

NOl-CB-74206 

Schwartz, Morton Memorial Hospital 

NOl-CB-53853 

Schwartz, Morton Sloan-Kettering 

NOl-CB-74086 

Sussman, Howard Stanford University 



IV. Morphologic Discriminants 

R23-CA-31755 

In Vitro Studies on Mammary Neoplastic Progression 
Asch, Bonnie Roswell Park Memorial Institute 

R23-CA-28128 

Tissue Blood Group Antigens and Carcinogenesis 
Howard, Donald Maine Medical Center 

*For all contracts, see respective contract summary. 



125 



NOl-CB-74098 

Page, David Vanderbilt University 

NOl-CB-74097 

Rosen, Paul Memorial Hospital 



EPIDEMIOLOGY 

I. Genetic Aspects of Susceptibility to Breast Cancer; Hormone 
Metabolism Patterns Associated with Familial High Risk 

ROl-CA-27632 

Genetic Epidemiology of Breast Cancer in Families 

King, Mary-Claire University of California, Berkeley 

ROl-CA-30069 

Estradiol Glucuronidation and Breast Cancer 
Zumoff, Barnett Montefiore Medical Center 



II. Steroid Hormones (Endogenous and Exogenous) and Their Contribution 
to Breast Cancer Risk; Hormonal Aspects of First Pregnancy; 
Epidemiologic Risk Factors in Relation to Levels of Steroid 
Hormone Receptors and of Estrogen-Binding 8-Globulin 

NOl-CB-53884 

Nomura, Abraham University of Hawaii 

NOl-CB-63996 

Deubner, David Duke University 

NOl-CB-74099 

Slone, Dennis/Shapiro, Samuel Boston University 

NOl-CB-74101 

Preedy, John Emory University 

POl-CA-13556 

Epidemiology and Natural History of Breast Cancer 

Petrakis, Nicholas University of California, San Francisco 

ROl-CA-28720 

Breast Cancer Biology: Epidemiology and Prognosis 
Deubner, David Duke University 

ROl-CA-30069 
Listed under I 

Zumoff, Barnett Montefiore Medical Center 



127 



III. Thyroid Function in Relation to Breast Cancer Risk. 

NOl-CB-84230 

Maloof, Farahe Massachusetts General Hospital 



IV. Epidemiology of Histopathologic Subcategories of Breast Cancer 

NOl-CB-53968 

Stenkvist, Bjorn University Hospital of Uppsala 

NOl-CB-63997 

Rosen, Paul Memorial Sloan-Kettering 



V. Epidemiology of Benign Breast Disease; Relation to Epidemiology 
and Natural History of Breast Cancer; Relation to Ethnicity and 
Risk of Breast Cancer 

NOl-CB-74102 

Modan, Baruch Chaim Sheba Medical Center 

NOl-CB-74202 

Spivey, Gary University of California, Los Angeles 

NOl-CB-84231 

Bartow, Sue University of New Mexico 

ROl-CA-26021 

Fibrocystic Breast Disease: Epidemiology and Histology 
Kelsey, Jennifer Yale University 

POl-CA-13556 

Listed under II 

Petrakis, Nicholas University of California, San Francisco 

ROl-CA-28223 

Epidemiologic Study of Breast Cancer Risk Factors 
Dupont, William Vanderbilt University 



VI. Epidemiology of Minimal Breast Cancer 

NOl-CB-74103 

Pasternack, Bernard New York University Medical Center 



128 



VII. Environmental Risk Factors for Breast Cancer (beside exogenous 
hormones) 

A. Diet (general; lipids and cholesterol; in relation to intesti- 
nal flora; in relation to hormone levels and metabolism; in 
relation to properties of the cell membrane; in relation to 
immune responsiveness and tumor cell susceptibility to 
cytotoxicity) 

NOl-CB-53884 

Nomura, Abraham University of Hawaii 

NOl-CB-74104 

Gorbach, Sherwood New England Medical Center 

NOl-CB-84229 

Kirschner, Marvin Newark Beth Israel Hospital 

NOl-CB-84318 

Papatestas, Angelos Mount Sinai School of Medicine 

ROl-CA-30273 

Dietary Fat Modulation of Mammary Tumorigenesis 
Erickson, Kent University of Calfornia, Davis 

ROl-CA-30629 

Promotion of Breast Cancer: Lipid Hormone Interactions 
Cave, William University of Rochester 

B. Possible Carcinogens and Mutagens in Breast Fluids 

POl-CA-13556 

Listed under II 

Petrakis, Nicholas University of California, San Francisco 



VIII. Correlation Between Breast Tumor Microscopic Characteristics and 
Patient Survival; Epidemiology, Morphology, Biochemistry, and 
Immunohistochemistry of Breast Tumors in Relation to Prognosis 

ROl-CA-28720 

Listed under II 

Deubner, David Duke University 

ROl-CA-30335 

Immunohistochemical Profile and Survival in Breast Cancer 
Lee, Arthur New England Medical Center 

ROl-CA-30342 

RNA Virus-Related Antigen in Breast Cancer and Prognosis 
Mesa-Tejada, Ricardo Columbia University 



129 



IX. Antiviral Antibodies and Breast Cancer; Relation to Ethnicity, toB 
Epidemiology and to Prognosis 

ROl-CA-29711 

Epidemiology and Etiology of Human Breast Cancer 
Day, Noorbibi Memorial Sloan-Kettering 

ROl-CA-30342 

Listed under VIII 

Mesa-Tejada Columbia University 



EXPERIMENTAL BIOLOGY 

I, Factors in the Induction of Mammary Tumors 

ROl-CA-18664 

Role of Prolactin in Mouse Mammary Tumorigenesis 

Sinha, Yagya Scripps Clinic and Research Foundation 

ROl-CA-27026 

Basis for Mammary Gland Susceptibility to Carcinogenesis 
Russo, Jose Michigan Cancer Foundation 

ROl-CB-27293 

Hormone Influences During Mammary Tumorigenesis ^i 

Socher, Susan Baylor College of Medicine 

ROl-CA-20764 

Neoplastic Mammary Gland: Structure/Function 
Strum, Judy University of Maryland 

ROl-CA-25915 

The Radiobiology of Mouse Breast Preneoplasia 

Cardiff, Robert University of California, Davis 

ROl-CB-31207 

Radioprotective Effect of Mammary Tumor Cell Grafts 
Scarantino, Charles Bowman Gray School of Medicine 

ROl-CA-21993 

Normal and Neoplastic Development of the Mammary Gland 

Dulbecco, Renato The Salk Institute for Biological Studies 

ROl-CA-22879 

Milk, Prolactin Binding and Thyroid in Breast Cancer 

Goodman, David Albany Medical College of Union University 

R23-CA-28999 

MMTV in Spontaneous and Carcinogen-Induced Tumors 

Pauley, Robert University of Miami ^ 



130 



ROl-CA-30036 

Mammary Metaplasia, Tumorigenesis and Chemoprevention 

Sorof, Sam The Institute for Cancer Research, Fox Chase 

ROl-CA-31774 

Estrogen Action in Normal Mammary Gland 

Haslam, Sandra Michigan State University 

ROl-CA-30570 

Modulation of Mammary Preneoplastic Progression 
Medina, Daniel Baylor College of Medicine 

ROl-CA-18946 

Cell Surface Modulation in Mammary Neoplasia 

Thompson, Karen Children's Hospital Medical Center 

NOl-CB-84227 

El-hawari, Monaem Midwest Research Institute, Kansas City 

NOl-CB-84226 

D'Ambrosio, Steven Ohio State University 



II. Growth Passage and Characterization of Normal Mammary Epithelial 
Cells 

POl-CA-05388 

Biology of Mammary Neoplasia 

Nandi, Satyabrata University of California, Berkeley 

ROl-CA-24844 

Characterization of Human Mammary Cells 

St^ampfer, Martha University of California, Berkeley 

ROl-CA-29090 

Characterization of MNU-Induced Mammary Cancers 

Grubbs, Clinton Southern Research Institute, Birmingham 

NOl-CB-74094 

Misfeldt, Dayton Stanford University 

NOl-CB-74188 

Masui, Hideo University of California, San Diego 



III. Mammary Tumor Response to and Dependence on Hormones 

ROl-CA-21606 

Role of Estrogen and other Hormones in Breast Cancer 
Butler, Barkley Michigan Cancer Foundation 



131 



ROl-CA-16660 

Insulin and Estrogen Interactions in Breast Cancer 
Hilf, Russell University of Rochester 



IV. Interaction of Neoplastic Mammary Cells with other Cell Populations 

ROl-CA-25179 

The Fibrotic Response to Human Breast Carcinoma 

Stern, Robert University of California, San Francisco 

ROl-CA-20764 
Listed under I 

Strum, Judy University of Maryland 

ROl-CA-28366 

Natural Site Preference in Mammary Cancer Biology 
Miller, Fred Michigan Cancer Foundation 

ROl-CA-20122 

Epithelial-Stromal Interactions of Breast Carcinoma 

Armstrong, Rosa University of California, San Francisco 

NOl-CB-84239 

Spring-Mills, Elinor State University of New York jr 



V. Cell Surface Proteins in Relation to Metastasis 

ROl-CA-19814 

Metabolic Fate of Mammary Cell Surface Glycoconjugates 
Bernacki, Ralph Roswell Park Memorial Institute 

ROl-CA-27909 

Adhesive Interactions in Mammary Tumor Epithelial Cells 
Buck, Clayton The Wistar Institute 

ROl-CA-31695 

Sialoglycoproteins of a Metastatic Mammary Tumor 
Carraway, Kermit University of Miami 

ROl-CA-19455 

Use of Membranes to Assess Phenotypic Expression 

Ceriani, Roberto Children's Hospital Medical Center 

ROl-CA-08418 

Glycoproteins from Cancer Cells 

Codington, John Massachusetts General Hospital 

ROl-CA-27314 

Plasma Membrane and Metastasis in Rat Mammary Cancer 
Fairbanks, Grant Worcester Foundation 



132 



ROl-CA-20605 

Mammary Cancer and Hormone Induced Responses 
Clark, James Baylor College of Medicine 

ROl-CA-30171 

Steroid Hormones in Breast Cancer 

Hockberg, Richard Yale University 

ROl-CA-26869 

Nuclear Estrogen Receptors in Breast Cancer 

Horwitz, Kathryn University of Colorado Medical Center 

ROl-CA-18664 
Listed under I 

Sinha, Yagya Scripps Clinic and Research Foundation 

ROl-CA-27293 
Listed under I 

Socher, Susan Baylor College of Medicine 

ROl-CA-28645 

Vitamin B6 and Hormone Action in Uteri and Breast Cancer 
Wotiz, Herbert Boston University School of Medicine 

ROl-CA-28698 

Perinatal Estradiol Influence on Mammary Development 
Warner, Marlene Baylor College of Medicine 

ROl-CA-28393 

Neonatal Hormone Exposure and Mammary Tumorigenesis 

Talamantes, Frank University of California, Santa Cruz 

ROl-CA-18458 

Fetal Exposure to Hormones and Mammary Carcinogenesis 
Boylan, Elizabeth Queens College of CUNY 

ROl-CA-16464 

lodinated Estrogens in Breast Cancer 

Caspi, Eluahu Worcester Foundation for Experimental Biology 

ROl-CA-22343 

Mechanism of Antiestrogen Action in Breast Cancer 

Chamness, Gary University of Texas Health Science Center 

ROl-CA-24687 

Metabolism of Estrogens and Androgens by Breast Cancer Cells 
Macindoe, John University of Iowa 

R23-CA-29501 

Estrogen Responsiveness in Human Breast Cancer 
Moore, Michael Marshall University 



133 



ROl-CA-26825 ^ 

Antigen Specific NK Activity and Suppression in MMTV 
Lane, Mary-Ann Sidney Farber Cancer Institute 

ROl-CA-25418 

Murine Virus Cross-Reacting Antigen in Human Tissue 

Hackett, Adeline University of California, Berkeley 

ROl-CA-28844 

Mammary Carcinoma Metastasis 

Nicolson, Garth University of Texas System Cancer Center 



VI. Prevention of Neoplastic Transformation 

ROl-CA-30036 
Listed under I 

Sorof, Sam The Institute for Cancer Research, Fox Chase 

NOl-CB-74207 

Moon, Richard IIT Research Institute 



VII. Pathophysiology of the Solid Mammary Carcinoma and its Relation 

to Metastasis ^ 

POl-CA-05388 

Listed under II 

Nandi, Satyabrata University of California, Berkeley 

ROl-CA-29537 

Osteotropism of Mammary Carcinoma Metastasis 

Mundy, Gregory University of Texas Health Science Center 

ROl-CA-28735 

Proteoglycans and Basal Structure and Function 
Bernfield, Merton Stanford University 



VIII. Immunodiagnosis and Immunoprevention of Cancer Growth and 
Progression 

ROl-CA-27437 

Immune Reactivity and Mammary Neoplasia 

Heppner, Gloria Michigan Cancer Foundation 

ROl-CA-30284 

An Immunodiagnostic Assay for Breast Cancer Detection 
Gaffney, Edwin The Pennsylvania State University 



134 



ROl-CA-20286 

Breast Neoplasia Diagnosis with Specific Antibodies 

Ceriani, Roberto Children's Hospital Medical Center 



XI. Service, Support and Resources 

R13-CA-30294 

Gordon Conference on Mammary Gland Biology 

Hilf, Russell University of Rochester Medical Center 

R13-CA-28816 

Manmary Cancer in Animals and Man - Conference 

Hageman, Philomena Netherlands Cancer Institute 

NOl-CB-74175 

Bogden, Arthur Mason Research Institute 

NOl-CB-84225 

Furthmayr, Heinz Yale University School of Medicine 

NOl-CB-84314 

Goldberg, Burton New York University Medical Center 



TREATMENT 

I. Steroid Receptors and Mammary Carcinoma 

A. Physiologic Behavior 

ROl-CA-27470 

Receptor Studies in Human Breast Cancer 

Hollander, Vincent Hospital for Joint Diseases 

ROl-CA-26452 

Studies of Anomalous Receptors in Breast Cancer 
Panko, Walter Baylor College of Medicine 

ROl-CA-22828 

Fate of Steroid Hormones in Breast Tumor Cells 

Brooks, Sam Wayne State University School of Medicine 

B. Methodology 

ROl-CA-29971 

Histochemical Methods for Receptors in Breast Cancer 

Chamness, Gary University of Texas Health Science Center 

NOl-CB-23862 

McGuire, William University of Texas Health Science Center 



135 



NOl-CB-43969 

Jensen, Elwood University of Chicago 

C. Evaluation of Clinical Application 

NOl-CB-04338 

Thomas, David Fred Hutchinson Cancer Research Center 



II. Biologic Markers in Breast Cancer 

A. Serum Resource, Storage and Distribution 

NOl-CB-74138 

Rodes, Ned Cancer Research Center, Columbia, MO 

NOl-CB-74213 

Bowman, Harold Butterworth Hospital 

NOl-CB-74137 

Whitney, Leslie Wilmington Medical Center 

NOl-CB -84296 

Go, Vay Liang Mayo Foundation 

G. Research Studies 

ROl-CA-26935 

CEA and FHAP as Breast Cancer Management Parameters 
Tormey, Douglass University of Wisconsin 

NOl-CB-43900 

Dao, Thomas Health Research, Inc. 

NOl-CB-74204 

Hilf, Russell University of Rochester 

III. Mechanisms of Treatment Action 

NOl-CB-84221 

Das Gupta, Tapas University of Illinois 

ROl-CA-26004 

Therapy Effects on Tumor Kinetics and Immune Parameters 
Fisher, Bernard University of Pittsburgh 

ROl-CA-25586 

Therapy-Induced Alterations in Breast Cancer 
Panko, Walter Baylor College of Medicine 



136 



ROl-CA-24129 

Control of Breast Cancer by Serum Growth Factors 

Osborne, Kent University of Texas Health Science Center 

ROl-CA-02071 

Estrogen Metabolism and Action in Pregnancy and Cancer 
Levitz, Mortimer New York University 

ROl-CA-30251 

Hormone Priming and Chemotherapy in Primary Breast Cancer 
Osborne, Kent University of Texas Health Science Center 

ROl-CA-23079 

Steroid Sulfurylation Inhibitors as Antitumor Agents 
Horwitz, Jerome Michigan Cancer Foundation 

ROl-CA-05197 

Endocrine Factors Influencing Tumor Growth in Man 
Pearson, Olof Case Western Reserve University 



IV. Preclinical Treatment Testing: Animal Models 

ROl-CA-29006 

C. Parvum Therapy of Mammary Carcinoma Metastases 
Kreider, John Pennsylvania State University 

ROl-CA-26287 

Canine Model for Human Breast Cancer Immunotherapy 
Hurvitz, Arthur The Animal Medical Center 



V. Clinical Investigations 

NOl-CB-43917 

Giuliano, Armando University of California, Los Angeles 

NOl-CB-43990 

Hubay, Charles Case Western Reserve University 

NOl-CB-33899 

Ahmann, David Mayo Foundation 

NOl-CB-53917 

Scanlon, Edward Evanston Hospital 

NOl-CB-53851 

Santen, Richard Pennsylvania State University 

ROl-CA-30006 

Chemotherapy and Anti-Estrogen Therapy in Breast Cancer 
Hubay, Charles Case Western Reserve University 



137 



VI. Data Management Resource 

NOl-CB-74140 

Dunsmore, Marlene Mason Research Institute 

NOl-CB-14339 

Dunsmore, Marlene Information Management Services, Inc. 



138 



CONTRACT RESEARCH SUMMARY 

Title: Estrogen Replacement after Premenopausal Oophorectomy and Breast 
Cancer Risk 

Principal Investigator: Dr. Dennis Slone, Dr. Samuel Shapiro 

Performing Organization: Boston University Medical Center 

City and State: Cambridge, MA 

Contract Number: N01-CB-74099 

Starting Date: 9/1/77 Expiration Date: 8/31/82 

Goal: To examine the hypothesis that exposure to female hormones, particularly 
estrogen replacement therapy, is related to an increased risk of breast cancer, 
either during use or after a latent interval. 

Approach: An epidemiological study of the case-control type is being imple- 
mented as part of an ongoing, hospital-based, multicenter data collection sys- 
tem. Cases of breast cancer and potential controls, women with a variety of 
non-malignant conditions, are interviewed by trained nurse monitors in hospi- 
tals throughout the country. Information on lifetime drug use and relevant 
medical data are recorded on standard forms and transferred to computer files 
for analysis. 

Progress: As of the end of September 1980, 1,685 women with breast cancer had 
been studied. Of these, 1,222 (73%) were newly diagnosed, of whom 110 (9%) 
were menopausal because of bilateral oophorectomy. Preliminary data do not 
provide any overall evidence that non-contraceptive estrogen use by women with 
premenopausal bilateral oophorectomy increases the risk of breast cancer. In 
the final analysis, the data will be stratified on risk factors for breast 
cancer, such as benign breast disease, parity and age at first pregnancy. As 
yet, numbers are insufficient to do this. Women with breast cancer are cur- 
rently being interviewed at a rate of 600 per year. 



Project Officers: Elizabeth P. Anderson, Ph.D., Robert Hoover, M.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: $36,000 



139 



CONTRACT RESEARCH SUMMARY 

Title: Biological Markers in Breast Cancer: Patient Resource 

Principal Investigator: Dr. Harold E. Bowman 

Performing Organization: Butterworth Hospital 

City and State: Grand Rapids, MI 

Contract Number: NOl-CB-74213 

Starting Date: 9/15/77 Expiration Date: 9/14/82 

Goal: To develop a Breast Cancer Task Force specimen resource for blood from 
breast cancer patients, benign disease patients, and controls to be used in a 
search for and verification of new breast cancer markers. 

Approach: Thirty milliliters of blood will be collected from breast disease 
patients who- are scheduled to undergo biopsy and/or primary surgery for breast 
lesions prior to surgery. Another specimen will be collected, when feasible, 
5-10 days postmastectomy from the same patient. Annual drawings are made on 
patients with malignant diagnosis. Patients with benign tumors are requested 
to complete annual questionnaires for a period of two years after biopsy. 
Serum specimens will be stored at -70°C, then shipped to an NCI-designated 
blood bank facility with appropriate clinical data. 

Progress: After a formal presentation to the surgery sections in each partici- 
pating hospital, surgeons who perform 95% of all breast biopsies in these hos- 
pitals signed letters of agreement allowing their patients to enter directly 
into the study. Since the inception of the program, 2,029 patients have been 
entered into the study; 435 of these patients have been found to have malignant 
breast disease. Approximately 26,000 vials containing serum specimens have 
been shipped to the central storage facilities at Mayo Clinic. 238 collections 
have been made on the annual anniversary of malignant patients and almost 1,000 
benign follow-up questionnaires have been completed on benign patients. This 
information has been forwarded to Mason Research computer bank. Fifteen months 
into the program, all participating patients and interested parties were in- 
vited to an informational update presentation provided by NCI project officers. 
The audience, numbering close to 500, were congratulated for their participa- 
tion and encouraged to continue their cooperative efforts. Participating sur- 
geons, pathologists, and hospital personnel working in the program attended a 
similar informational session. 



Project Officers: Ihor J. Masnyk, Ph.D., Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: $46,550 



140 



• 



CONTRACT RESEARCH SUMMARY 

Title: Biologic Markers in Breast Cancer: Patient Resource 

Principal Investigator: Dr. Ned D. Rodes 

Performing Organization: Cancer Research Center 

City and State: Columbia, MO 

Contract Number: NOl-CB-74138 

Starting Date: 9/1/77 Expiration Date: 8/31/82 

Goal: To serve as a Breast Cancer Task Force specimen resource for blood from 
breast cancer patients and controls to be used in a search for and verification 
of new breast cancer markers. 

Approach: Thirty milliliters of blood are collected from volunteer" Breast Can- 
cer Detection Project and Women's Cancer Control Program screenees after they 
have signed appropriate consent forms. In the event of subsequent breast biop- 
sies upon any of the participants, a 30 ml postoperative specimen will also be 
collected from postmastectomy cases within three months. In addition, prebi- 
opsy and postmastectomy blood samples will be obtained from hospitalized ladies 
in the Colximbia area who agree to participate in the program. Emphasis will 
be placed upon the collection of samples from hospitalized patients as opposed 
to specimen collection from normal volunteer screenees. No special require- 
ments will be placed upon the participants in terms of diet or liquid intake. 
Anniversary serum from malignant patients will be obtained for three, possibly 
five years. All blood will be stored at -70°C, and then shipped to NCI-desig- 
nated blood bank facility with appropriate clinical data. 

Progress: In 40 months of serum collection (from 12/77 to 4/81), 11,379 indi- 
vidual samples of blood were fully processed; this number of blood samples 
yielded 122,080 vials. The average serum collection per participant has been 
10.0 ml. The average amount of serum per vial was 1.1 ml. The total number 
of "voids" (no draws; broken clot tubes; collapsed veins; hemolysis; etc.) was 
limited to 426 over the entire collection period. The average acceptability 
for research of shipments during this time frame was 96.2%. Surgeons at the 
Ellis Fischel State Cancer Hospital have agreed to allow our blood collection 
team access to their hospitalized, pre-breast biopsy patients. Three more hos- 
pitals in the area are currently bringing this matter under administrative 
review. If they agree to cooperate, all the non-federal hospitals in the 
Colinnbia area will be indirect participants in this NCI study. 



Project Officers: Ihor J. Masnyk, Ph.D., Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: ^52,250 



141 



CONTRACT RESEARCH SUMMARY 

Title: Therapy for Stage II or Stage III Carcinoma of the Breast 

Principal Investigator: Dr. Charles A. Hubay 

Performing Organization: Case Western Reserve University 

City and State: Cleveland, OH 

Contract Number: NOl-CB-43990 

Starting Date: 6/16/74 Expiration Date: 6/15/82 

Goal: This clinical trial was initiated in September 1974, to determine if 
combined chemotherapy (CMF), anti-estrogen therapy (Tamoxifen) and immunother- 
apy (BCG) following mastectomy in Stage II, III breast cancer would yield bet- 
ter results than when used later in the course of the disease when recurrence 
became manifest. 

Approach: Patients under age 76 who show axillary nodes involved with metasta- 
ses at the time of surgery were eligible for this study. Stratification, but 
not treatment selection, was on the basis of the presence or absence of estro- 
gen receptor protein (ER) in the tumor. Random treatment assignments were (1) 
cyclophosphamide, methotrexate, 5-f luorouracil (CMF); (2) cyclophosphamide, 
methotrexate, 5-f luorouracil. Tamoxifen (CMFT); and (3) CMFT for 12 months plus 
BCG for 12 months. Endpoint was the first evidence of treatment failure, i.e., 
the appearance of local recurrence or of distant tumor. 

Progress: This study was closed in June 1979. Three hundred eighteen patients 
were enrolled in the study. Of these, 76% had estrogen receptor (ER) positive 
tumors (_> 3 f emtomoles/mg of protein) . All patients have completed one year of 
chemotherapy. Twenty patients are completing their 12 months of BCG therapy. 
Of the treated patients, 48 have died and 96 have had treatment failure. 
Thirty-three patients have withdrawn voluntarily after one month to one year 
of therapy. Chemotherapy was not accompanied by significant myelosuppression. 
Hair loss was slight. Tamoxifen, the antiestrogen agent used, was tolerated 
well, without serious side effects, although most patients experienced hot 
flashes while on the drug. Ninety-six patients have had recurrence; 35 in the 
CMF group, 30 in the CMF + Tamoxifen, and 31 in the CMF + Tamoxifen + BCG group. 
Statistical comparison for treatment failure between Stage II ER positive and 
ER negative patients is significant at the p <. 0.0001 level, with the latter 
group recurring more rapidly (48.6% vs. 38.8% at 48 months). Relapse rate for 
the Stage II treated patients at 48 months is as follows: CMF 44.5%, CMF + 
Tamoxifen 40.0% and CMF + Tamoxifen + BCG 40.4%. Recurrence rates between Stage 
II patients with 1-3 positive nodes are significantly different when stratified 
according to estrogen receptor values, with higher relapse rates in the ER nega- 
tive group. 

Stratification by estrogen receptor assay shows a significant difference in 
recurrence when Tamoxifen is added to CMF therapy in ER+ patients (p = 0.03). 
There is no difference in recurrence rates for ER negative patients with any of 
the three treatments used. Survival data at 48 months mean follow-up show a 
significant mortality for the ER negative patients vs. ER+ patients (p < 0.0001), 

Project Officer: Mary E. Sears, M.D. 

Program: Breast Cancer Treatment > 

FY 81 Funds: $75,000 



142 



• 



CONTRACT RESEARCH SUMMARY 

Title: Epidemiology of Benign Breast Disease 

Principal Investigator: Dr. Baruch Modan 

Performing Organization: Chaim Sheba Medical Center 

City and Country: Tel Hashomer, Israel 

Contract Niimber: NOl-CB-74102 

Starting Date: 8/1/77 Expiration Date: 7/31/81 

Goal: To determine the characteristics of patients with different types of 
noninvasive breast lesions, with particular reference to diet and to those 
factors reputed to be preferentially associated with breast cancer, and to 
compare the findings with our similarly conducted breast cancer study. 

Approach: The study consists of two phases: (a) case-control interview study 
of 600 cases of noninvasive breast diseases in terms of such risk, factors as: 
dietary habits, age at first birth, menstrual history, menopausal status, par- 
ity, height and weight, familial breast cancer, selected drug history, and pre- 
vious radiation exposure; and (b) an incidence study of noninvasive breast le- 
sions in a total community. The characterization of the breast lesions will 
take account of the histological pattern, degree of atypia, and lymphoreticulo- 
endothelial response (LRE). Particular attention will be paid to the relative 
frequency of distinct histopathological forms in highand low-risk subpopula- 
tions (i.e., Europeans, Asian-African born). 

Progress: As of November 1980, 2,180 interviews have been conducted: 874 cases, 
709 surgical controls and 597 neighborhood controls. All the interviews have 
been performed at home. All of the histopathological slides have been read by 
both study pathologists. The frequency distributions of cases by conventional 
pathological criteria are as follows: fibroadenoma 179, proliferative 361, 
microcystic 117, intraductal papilloma 14, combined histology 53, in situ carci- 
noma 18, nonspecific histopathology 82, cystosarcoma 7, granuloma 46; according 
to Black's gradings: Grade 1: 361, Grade 1-2: 3, Grade 2: 204, Grade 2-3: 
131, Grade > 3: 95. Previous surgery has been validated. The incidence study 
of the whole spectrum of breast disease began in July 1979, and collection of 
slides and cases has been concluded. The study includes demographic and patho- 
logical data; 80% of the histological slides have been reviewed using the Black 
classification. Approximately 4,200 cases have been included up to now. 



Project Officer: Elizabeth P. Anderson, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



143 



CONTRACT RESEARCH SUMMARY 

Title: Epidemiologic Characteristics of Pre- and Postmenopausal Breast Cancer ^ 

Principal Investigator: Dr. David C. Deubner 

Performing Organization: Duke University Medical Center 

City and State: Durham, NC 

Contract Number: NOl-CB-63996 

Starting Date: 6/30/76 Expiration Date: 1/31/81 

Goal: To define the epidemiologic characteristics of breast cancer in pre- and 
postmenopausal women, and of breast cancers rich and poor in cytoplasmic recep- 
tors for estrogen and progesterone. 

Approach: This case-referent study is of breast cancer patients undergoing 
surgery at university and -community hospitals, surgical and medical patients 
not having breast or gynecological surgery at the same hospitals, and subjects 
from the community. Data collection is by interview of all subjects and review 
of charts of hospital patients. Laboratory studies on tissue and serum from 
breast cancer patients include quantitative estrogen and progesterone receptor 
determinations, evaluation of histology and ultrastructure by a standardized 
protocol, and estrogen and progesterone serum determinations. Statistical anal- 
yses estimate the relative strength of association between breast cancer in 
pre- and postmenopausal women and such factors as age of menarche, age of first 
delivery, obesity, and family history of breast cancer. Associations are also 
being sought between these factors and concentrations of the cytoplasmic hor- 
mone receptors. ■ 

Progress: As of November 1980, collection and computer entry of interview, 
hospital chart review, and estrogen receptor data was complete. Light micro- 
scopy review was complete and ultrastructure review well under way. Analyses 
of interview data relevant to the epidemiology of pre- and postmenopausal 
breast cancer, and to the epidemiology of estrogen receptors, are also complete 
and are being prepared for publication. 



Project Officers: Elizabeth P. Anderson, Ph.D., Gordon B. Cutler, M.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



144 



CONTRACT RESEARCH SUMMARY 

Title: Biologic Characterizations of "Premalignant" Human Mammary Epithelial 
Hyperplasias 

Principal Investigator: Dr. Kenneth Scott McCarty, Jr. 

Performing Organization: Duke University 

City and State: Durham, NC 

Contract Number: NOl-CB-84223 

Starting Date: 8/1/78 Expiration Date: 7/31/81 

Goal: To characterize potentially "premalignant" epithelial lesions of the 
human breast using biochemical, morphologic and immunologic techniques. 

Approach: Analyses are performed on lesions observed in subcutaneous mastec- 
tomy specimens and benign biopsies. These are compared to carcinomatous areas 
of breast cancer specimens. Lesions are classified by subgross, light, and 
electron microscopic techniques as appropriate. The tissues are evaluated for 
the presence of measurable steroid receptors, estrogen-progesterone binding 
proteins, endogenous peroxidase, K-casein, and CEA. Immunologic characteriza- 
tions include immunohistologic assessment of IgG, IgA, IgM and the expression 
of ABH blood group antigens and/or the unmasking of component antigens. 

Progress: The effects of the menstrual cycle on the normal and fibrocystic mam- 
mary gland have been characterized and categorized into five phases: 1) proli- 
ferative (days 3-7), 2) follicular phase of differentiation (days 8-14), 3) 
luteal phase of differentiation (days 15-20), 4) secretory (days 21-27), and 5) 
menstrual (days 28-2). A statistically significant association of the simulta- 
neous presence of certain patterns of intraacinar epithelial hyperplasias and 
carcinoma has been confirmed. Apocrine and adenosquamous differentiation have 
been characterized ultrastructurally. Attenuation of the epithelial stromal 
junction was associated with IgG localization and alteration in the ABH blood 
group antigen expression in mammary dysplasias. CEA is seen in malignant le- 
sions and severe epithelial dysplasias but only rarely (< 5%) in the simple 
fibrocystic lesions. K-Casein does not appear to correlate with E2 receptor. 
The localization of immunoglobulin G in a pericellular pattern has been observed 
in dysplastic epithelial hyperplastic lesions (usually type 14 positive), while 
apical IgA localization has characterized the non-dysplastic benign lesions 
studied. IgG localization correlates with lymphopenia. FITC labeled steroids 
to localize sex steroid binding have suggested that 17 substituted compounds 
correlate best with SDGA techniques, although E2-FITC binding to low-affinity 
high-capacity proteins is likely due to the correlation of binding to the pres- 
ence of estrogen-progesterone binding protein. Menstrual cycle fluctuations of 
estrogen and progesterone receptor have been observed in benign, dysplastic and 
malignant lesions. The levels of intermediate affinity estrogen-progesterone 
binding protein are maximal in the late luteal phase. 



Project Officer: D. Jane Taylor, Ph.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



145 



CONTRACT RESEARCH SUMMARY 

Title: Endocrine Events at the Time of First Pregnancy 

Principal Investigator: Dr. John R. K. Preedy 

Performing Organization: Emory University School of Medicine 

City and State: Atlanta, GA 

Contract Number: NOl-CB-74101 

Starting Date: 8/1/77 Expiration Date: 7/31/82 

Goal: It is proposed that the protective effect of an early first pregnancy 
against breast cancer is due to a change in the hormonal environment following 
that event. We proposed to investigate and characterize such a change. 

Approach: Nulliparous subjects in the 18-22 age range who have not taken birth 
control pills, who have a history of normal regular menses, and who are plan- 
ning pregnancy in the next 1-2 years will be selected (Group A). Similar sub- 
jects will be chosen who do not plan to become pregnant in the next 1-2 years 
(Group B). Similar subjects to the above but in the age range 30-40 would also 
be selected (Groups C and D). All groups would have the following carried out 
in the early follicular phase of the menstrual cycle: plasma gonadotropins, 
estrogens, progesterone, androgens; urine estrogens and androgens; plasma pro- 
tein-steroid binding studies; perphenazine-prolactin stimulation test; LRH-LH 
stimulation test. Results from Groups A, B, C, and D would be compared by 
appropriate analysis of variance techniques to determine significant contrasts 
among the four groups. 

Progress: The total number of subjects participating in the program is 122. 
Group A - 41 (Initial study has been done on all 41. Twenty have become preg- 
nant and four of these have suffered miscarriage. Fourteen have reached full 
term and delivered and the remaining two are still pregnant. Four of the sub- 
jects who have delivered have been restudied) . Group B - 29 (All 29 have 
undergone the initial study and ten have been restudied). Group C - 24 (All 
24 have undergone the initial study. Ten have become pregnant and one of these 
has suffered a miscarriage. Nine have so far delivered and one of these has 
been restudied). Group D - 28 (All 28 have undergone the initial study and 
nine of these have been restudied). 



Project Officer: Elizabeth P. Anderson, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: $27,000 



146 



CONTRACT RESEARCH SUMMARY 

Title: Therapy of Patients with Stage II or Stage III Carcinoma of the Breast 

Principal Investigator: Dr. Edward F. Scanlon 

Performing Organization: Evanston Hospital 

City and State: Evanston, IL 

Contract Number: NOl-CB-53917 

Starting Date: 6/30/75 Expiration Date: 6/29/82 

Goal: To determine the efficacy of adjuvant chemotherapy and chemoimmuno ther- 
apy for patients who have had standard surgical intervention for Stage II or 
III carcinoma of the breast. 

Approach: Patient accrual began in July 1975, and was completed as of July 
1979. A total of 194 patients were entered into the study during this time. 
Stratification and randomization were carried out along with a balancing for 
prognostic factors according to the following variables: primary tumor size, 
number of positive nodes, menopausal status, and unfavorable local signs. The 
statistical analysis involved sequential treatment assignment and was designed 
to provide the greatest balance among the three groups. Using this method, 
patients were originally assigned to one of three treatment schedules: Group 
I - Oral phenylalanine mustard (L-Pam), Group II - 5-Fluorouracil, cyclophos- 
phamide and prednisone (CFP), and Group III - CFP plus BCG inoculations. 
Enrollment in the single arm treatment. Group I, was discontinued in November 
1977 because of a disproportionately high recurrence rate in the group at that 
time. The 38 patients already enrolled in that group at that time. The 38 
patients already enrolled in that group remained to finish their single arm 
treatment. New patients from that time were enrolled either in Group II or III. 

Progress: Fifty-six patients have experienced recurrent disease. Fifteen of 
the 38 patients in the L-Pam group (39%) report recurrences. Of the 78 patients 
in the CFP group, 22 (28%) report recurrences. Of the 78 patients in the third 
group, CFP + BCG, 19 (24%) report recurrent disease. At the time enrollment was 
discontinued in the L-Pam group, a higher recurrence rate was observed in that 
group than in the polychemotherapy groups. Since that time, the recurrence rate 
has dropped for the single arm treatment group, and the recurrence rate for the 
polychemotherapy groups has steadily increased. Currently, no statistically 
significant difference exists among the three treatment groups with regard to 
disease-free interval. Two prognostic factors, tumor size (_> 3 cm) and nodal 
involvement (> 4), have emerged as statistically significant in terms of progno- 
sis. We will be continuing to follow these patients very closely over the next 
few years in order to determine disease-free interval and, ultimately, survival. 
Data are also currently being combined with data from the Mayo Clinic study, 
which is similar in design to this project, in order to provide a larger patient 
population for analysis. 



Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: $36,000 



147 



CONTRACT RESEARCH SUMMARY 

Title: Evaluation of the Impact of the Estrogen Receptor Assay on the 
Treatment of Human Breast Cancer 

Principal Investigator: Dr. David B. Thomas 

Performing Organization: Fred Hutchinson Cancer Research 

Center 
City and State: Seattle, WA 

Contract Number: NOl-CB-04338 

Starting Date: 9/1/80 Expiration Date: 6/30/82 

Goal: To identify characteristics of women, their physicians and hospitals 
that distinguish individuals with breast cancer who have steroid hormone recep- 
tor assays performed on samples of their primary or recurrent tumor tissue; and 
to determine whether therapy for primary or recurrent disease is altered by the 
assay results. 

Approach: Women with primary breast cancer in 1977-78 and 1980, and women in 
the former group who develop recurrent disease by the end of 1981 are identi- 
fied through the Cancer Surveillance System (CSS) which is a population-based 
tumor registry that has been used to collect data on all newly diagnosed cancer 
cases in a 13-county area of Washington State since 1974. Descriptive informa- 
tion on all study subjects is being collected from the registry and this is 
supplemented by data on some women with disease onset in 1977-78 that were col- 
_lected by personal interviews as part of a previous study. Information on ste- 
roid receptor assays is collected from medical records. Information on physi- 
cians is obtained from professional directories and a state survey of physi- 
cians; and data on hospitals are abstracted from similar sources. Histologic 
and clinical characteristics of the tumor and types of therapies given are ob- 
tained from CSS records and will be supplemented by querying physicians. Assay 
methods used and quality control procedures followed will be ascertained from 
laboratory directors. 

Progress: A total of 1,206 female residents of King County who developed breast 
cancer in 1977 or 1978 were identified. Information on steroid assays, recur- 
rences of disease, and therapy for recurrent tumors is currently being col- 
lected. Women who are residents of the 13-county catchment area and who develop 
breast cancer in 1980 are being identified as they are detected by registry per- 
sonnel and information on steroid receptor assays is being collected at the same 
time that routine data for the registry are being collected as physicians are 
identified through the registry. Data on all area hospitals have been obtained. 



Project Officer: Mary E. Sears, M.D. 

Program: Breast Cancer Treatment 

FY 81 Funds: ($105,073 funded by NIH-OD) 



148 



• 



CONTRACT RESEARCH SUMMARY 

Title: Steroid Sulfation and Estrogen Binding in Human Breast Cancer 

Principal Investigator: Dr. Thomas L. Dao 

Performing Organization: Health Research, Inc. 

City and State: Buffalo, NY 

Contract Number: NOl-CB-43900 

Starting Date: 2/1/74 Expiration Date: 1/31/82 

Goal: To determine the relationship of steroid sulfotransf erase activity to 
estrogen receptor protein levels, to pathologic staging, and to risk of relapse. 

Approach: Steroid sulfating enzyme activity and estrogen receptor (ER) levels 
have been assayed in breast tissue obtained from approximately 300 patients in 
three institutions. The evidence will be evaluated for or against correlation 
of the steroid sulfotransf erase activity with risk for tumor metastasizing po- 
tential and with histopathologial parameters. The predictive potentials of ER 
and sulfation activity will be compared. 

Progress: A total of 266 patients from three cooperating institutions were en- 
tered into the study between March 1974 and June 1976, inclusive. The average 
length of follow-up is now approximately 56 months. The data have been analyzed 
according to the following categories: DHEAS versus other prognostic indica- 
tors; E2S versus other prognostic indicators; ER versus other prognostic indi- 
cators; and the recurrence rate for categories of prognostic indicators. The 
prognostic indicators considered were: age at diagnosis, pathological size of 
the tumor, histologically positive axillary nodes, and menopausal status. The 
synthesis of sulfotransf erase was also evaluated for its correlation with age 
at diagnosis, tumor size, axillary nodal metastasis, and menopausal status. 
However, because of size, ER and sulfotransf erase activities could not be mea- 
sured in all tumors. The present analysis includes only those patients for whom 
both measurements were done. The data revealed that (1) DHEAS and E2S are 
highly correlated with each other. Although there is a trend for DHEAS to 
increase with the size of the tumor, this association is not statistically sig- 
nificant. (2) ER levels are significantly correlated with the patient's age at 
diagnosis, the size of the tumor, and menopausal status. The ratio of DHEAS + 
I/E2S + 1 is significantly correlated with ER levels; ER levels and this ratio 
are negatively correlated. (3) The ratio of DHEAS + I/E2S + 1 is signifi- 
cantly correlated with the recurrence rate, and appears to be an independent 
variable. The recurrence rate is significantly lower in patients whose tumors 
have a ratio of < 1.5, whereas in contrast, the recurrence rate is higher in 
patients having a ratio of >1.5. This is true in patients having no nodal 
metastasis, as well as in those having 4+ nodes involved. 



Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: ilO,000 



149 



CONTRACT RESEARCH SUMMARY 

Title: Prevention of the Formation and Progression of Mammary Gland 
Preneoplastic Lesions 

Principal Investigator: Dr. Richard D. Moon 

Performing Organization: IIT Research Institute 

City and State: Chicago, IL 

Contract Number: NOl-CB-74207 

Starting Date: 8/29/77 Expiration Date: 2/28/81 

Goal: To study the use of retinoids to prevent the development and/or progres- 
sion of preneoplastic lesions of the mammary gland. 

Approach: Experimental animals are fed nontoxic levels of synthetic vitamin A 
analogues (retinoids) as a dietary supplement in the attempt to inhibit the 
development and progression of preneoplastic lesions in the mammary glands of 
mice and carcinogen-treated rats. Endpoints monitored are: incidence of 
hyperplastic alveolar nodules (HAN) and "spontaneous" mammary tumors in C3H/He 
mice, and incidence, latency, and multiplicity of chemical carcinogen-induced 
mammary tumors in Sprague-Dawley rats. The effects of beginning retinoid 
administration at various times after carcinogen administration are being exam- 
ined, as is the influence of retinoid administration combined with bilateral 
ovariectomy of carcinogen-treated animals. 

Progress: Previous studies have shown that combination treatment of the reti- 
noid and a prolactin suppressor was more effective in inhibiting MNU-induced 
mammary carcinogenesis than either treatment alone. Thus, studies were initi- 
ated to determine the effect of retinoids on prolactin-induced mammary gland 
differentiation in organ culture. Both all-trans-retinoic acid and 4-hydroxy- 
phenyl retinamide (4-HPR) inhibited lobular development and reduced the number 
of ductal branchings. Also, 4-HPR exerted a dose-related inhibitory effect on 
mammary parenchymal cell DNA synthesis. Furthermore, other data indicated that 
the inhibitory effect of 4-HPR on prolactin-induced mammary gland differentia- 
tion may be mediated by retinoic acid binding proteins. In other studies in 
which a 100% MNU-induced mammary tumor incidence with more than 4 tiimors per 
rat was attained, ttmior incidence in ovariectomized, 4-HPR-treated animals was 
0. Thus it appears that 4-HPR is highly effective in inhibiting the induction 
of ovarian hormone independent mammary cancers. Moreover, the combination 
treatment of ovariectomy and retinoid was also highly effective in inhibiting 
the appearance of a second tumor when retinoid treatment was initiated follow- 
ing the surgical removal of the first tumor. Additional studies in pregnant 
and lactating rats indicate that the levels of retinoid receptor in the mammary 
tissue may be directly related to the circulating levels of estradiol in the 
animal. 



Project Officer: Chester V. Piczak, B.S., D. Jane Taylor, Ph.D. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



150 



CONTRACT RESEARCH SUMMARY 

Title: Biomedical Computing Software Support of Breast Cancer Treatment 
Program 

Principal Investigator: Ms. Marlene Dunsmore 

Performing Organization: Information Management Services, Inc. 

City and State: Bethesda, MD 

Contract Number: NOl-CB-14339 

Starting Date: 3/31/81 Expiration Date: 3/30/82 

Goal: To increase the usefulness of the data generated in projects sponsored 
by the Breast Cancer Task Force Treatment Program and in other designated pro- 
jects related to the treatment of human breast cancer. 

Approach: A central file was developed by the initial contractor for three 
areas of study: (1) surgical adjuvant therapy, (2) tumor biomarkers, and (3) 
estrogen receptor assays. The file allows comparisons of the results from 
various studies and provides a data base from which material can be quickly and 
conveniently retrieved. This data file is also intended for testing new ideas, 
identifying groups of subjects suitable for more detailed study, and for pre- 
paring reports to the medical community and the general public. The file is 
not intended to duplicate those at individual institutions, but to prepare data 
for analyses that are not possible at the individual institutions. The current 
contractor will continue this data file. 

Progress: The transition of the Breast Cancer files and belongings to I. M.S. 
went smoothly. Communication with the collaborating institutions has been 
maintained. 



Project Officers: Mary E. Sears, M.D., Ihor Masnyk, Ph.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: $149,445 



151 



CONTRACT RESEARCH SUMMARY 

Title: Isoproteins in Normal, Benign and Malignant Breast Tissues 

Principal Investigator: Dr. Doris Balinsky 

Performing Organization: Iowa State University- 

City and State: Ames, lA 

Contract Number: NOl-CB-84222 

Starting Date: 8/1/78 Expiration Date: 7/31/81 

Goal: To detect, quantitate, and characterize various isozymes or isoproteins 
in normal, benign and neoplastic breast tissue in an attempt to identify "tumor 
markers" which may have significance in the early detection of breast cancer 
or identification of patients with occult dissemination after local treatment. 

Approach: A number of enzymes known to have differing isozyme patterns in dif- 
ferent tissues will be assayed in human normal, benign, malignant and metastatic 
breast tissues. Isozyme patterns of those enzymes having detectable activity 
will be examined by electrophoresis. Subsequently, those isozymes showing dif- 
ferences between normal and malignant tissues will also be examined in sera from 
these patients. If interesting isozyme differences are found between normal and 
cancerous tissues, the isozymes will be purified and characterized. These stud- 
ies could indicate altered metabolic status of the tumor. The data will be cor- 
related with data on estrogen and progesterone receptors on the same material 
and with clinical and histopathologic data. 

Progress: Twenty-three different sets of isozymes have been examined to date 
using cellulose acetate electrophoresis. The isozjnne patterns of lactate dehy- 
drogenase (LDH), malate dehydrogenase (MDH) and the esterases appear the most 
promising as tumor markers. LDH5 generally predominated in breast carcinoma, 
while LDH3 was the major isozyme in normal breast and fibrocystic disease, and 
in ductules derived from normal breast by collagenase digestion. One-fourth of 
the malignant tumors had a predominance of the mitochondrial form of MDH, while 
the cytosolic form predominated in all the normal and benign tissues. Acetyles- 
terases A4 and A5 were low or absent in all the normal tissues, but were a 
major component of many of the tumor tissues, both malignant and benign. Butyr- 
ylesterase B3 was a major component of most of the tissues examined, but was 
low in a few of the malignant tissues. No correlations of enzyme pattern with 
steroid receptors, clinical or pathological status were found. 



Project Officer: D. Jane Taylor, Ph.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



152 



• 



CONTRACT RESEARCH SUMMARY 
Title: Animal and Human Mammary Tumors and Human Cell Culture Bank. Facility- 
Principal Investigator: Dr. Arthur E. Bogden 
Performing Organization: Mason Research Institute 
City and State: Worcester, MA 

Contract Number: NOl-CB-74175 

Starting Date: 6/28/79 Expiration Date: 6/27/81 

Goal: To function as a service facility for the cryopreservation, storage, and 
distribution of biologically characterized and monitored (a) animal and human 
mammary and endocrine related tumors transplantable in_ vivo , and (b) cell lines 
of human and animal breast tumor origin established in in vitro culture, for 
use by the Breast Cancer Task Force and other selected investigators. 

Approach: Cryopreservation of mammary and endocrine related tumors of animal 
and human origin, as well as human breast tumor cell lines: (a) submittal of 
tumors and cell lines by qualified investigators able to furnish pertinent 
background information; (b) biological characterization of transplantable tis- 
sues, both pre- and post-cryopreservation, by determination of growth curves, 
specific effects on host organs, host (syngeneic and xenogeneic) survival, 
serum hormone levels, histology, karyology, response to ablative procedures; 
(c) cell lines tested for viability, plating efficiency, freedom from contami- 
nation and tumorigenicity by nude mouse implantation; (d) characterized tissue 
and cell lines preserved in Linde liquid nitrogen freezers according to well 
recognized, proven procedures. 

Progress: During the period December 1, 1979 to December 1, 1980, 184 tumors 
were shipped to 91 investigators; 41% were of mammary and 8% of anterior pitui- 
tary origin. Two new timiors were received for cryopreservation and characteri- 
zation and 9 human tumors (non-breast) were transferred to the DCT Tumor Bank. 
Breast tumor cell lines DU4475 and MDA-MB-436 have been established as trans- 
plantable solid tumor systems in athymic nudes. There is a current inventory 
of 252 transplantable tumors or sublines stored in 5,130 ampules in liquid N2. 
Eight shipments of hximan and rat a-lactalbumin and/or specific antisera have 
been made. Anti-Type I, II and III collagen antiserum was received and is now 
also available for distribution. Current inventory of cell lines in cryopreser- 
vation in the in vitro cell bank includes 20 human breast tumors, 4 human normal 
epithelial, and 2 mouse and 1 rat mammary tumors. One hundred and eight ship- 
ments, which included 14 frozen ampules and 410 viable cultures, were made to 
74 different laboratories, 16 of which were in foreign countries. During the 
past year there has been a 23% increase in shipments of i_n vivo animal tumor 
systems and a 74% increase in shipments of in vitro human cell lines. A biblio- 
graphy relative to the transplantable tiamors is being distributed upon request. 



Project Officers: Chester V. Piczak, B.S., D. Jane Taylor, Ph.D. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: $148,500 



153 



CONTRACT RESEARCH SUMMARY 
Title: Data Research Analysis for Breast Cancer Treatment Program 



Principal Investigator: 
Performing Organization; 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-74140 
3/21/77 



Ms. Marlene Dunsmore 
Mason Research Institute 
Rockville, MD 



Expiration Date: 3/20/81 



Goal: To increase the usefulness of data produced in projects related to the 
treatment of human breast cancer. 

Approach: A central data file has been developed for three areas of study: 
(1) surgical adjuvant therapy; (2) tumor markers; and (3) estrogen receptor 
assays. The file allows comparisons of the results from various studies and 
provides a data base from which material can be quickly and conveniently 
retrieved. This data file is also implemented for testing new ideas, identify- 
ing groups of patients suitable for more detailed study, and for preparing 
reports to the medical community and the general public. The file is not in- 
tended to duplicate those at individual institutions but to perform analyses 
that are not possible at the individual institutions. 

Progress: Mason personnel have designed a data collection and editing system 
for the Breast Cancer Studies Data Center. Clinical information is abstracted 
and entered via an IBM on-line disk file. The main file update system modifies 
clinical history files with new information and continues to edit for data con- 
sistency. Data concerned with treatment, histopathology, survival, and estro- 
gen receptor status have been stored on 2,804 patients. Personnel have pre- 
pared extensive tabulations of these data for general meetings of the Breast 
Cancer Task Force Committee. They have sent each participating institution an 
analysis of its own data and of the collated information. 

In February 1978 work was begun in support of the Biological Markers Pro- 
ject. The basic data collection and editing systems are operative. Background 
and clinical data have been gathered on 14,349 patients from three institutions. 
Mayo Clinic is serving as the repository for the serum samples. Benign tumor 
and metastatic cancer follow-up information is now being gathered. Research 
investigators have so far requested 8 shipments of serum specimens to test spe- 
cific blood markers. Their laboratory results are being returned to the Data 
Center for evaluation. 



C 



i 



Project Officers: Mary E. Sears, M.D., Ihor J. 
Program: Breast Cancer Treatment 
FY 81 Funds: 



Masnyk, Ph.D. 



154 



CONTRACT RESEARCH SUMMARY 
Title: The Relationship Between Thyroid Disease and Breast Cancer 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Farahe Maloof 
Massachusetts General Hospital 
Boston, MA 



Contract Number: 
Starting Date: 



NOl-CB-84230 
9/30/78 



Expiration Date: 9/29/82 



Goal: To examine the possible association of breast cancer with prior thyroid 
dysfunction. 

Approach: This is a retrospective cohort study designed to measure the rates 
of breast cancer in women with and without prior thyroid disease. A mortality 
study will follow 7,000 women with thyroid disease seen at the Thyroid Clinic 
of the Massachusetts General Hospital between 1925 and 1975, and 7,000 matched 
population controls; 2,000 women seen at the Clinic but without identified thy- 
roid disease will also be followed. Rates will be compared in women with vari- 
ous types of thyroid disease. A morbidity study will follow 1,600 women with 
previous thyrotoxicosis; this will include 500 women who became hypothyroid 
following the treatment and 500 who did not. Rates of breast disease will be 
compared according to mode of treatment as well as externally compared with 
rates from the Connecticut Tumor Registry. 

Progress: Mortality Study: The card file of patients treated at the Thyroid 
Clinic was reviewed. From the 45,000 names, 13,400 women satisfied the study 
criteria, almost twice as many women as had been anticipated, allowing us to 
increase the sample size. This included 3,140 women with no thyroid disease. 
Pertinent medical data and follow-up information have been abstracted from the 
medical records of 9,600 of these women. One control woman was sought from the 
available Massachusetts Residents' List for each thyroid patient who resided 
in the state at the time of diagnosis; 10,200 women who resided in the same 
neighborhood and who were within two years of age of the case were selected. 
The information present in the Resident's List was abstracted, keypunched, and 
entered into the computer for each control woman selected. We recently began 
the follow-up search of the Massachusetts death certificates. So far, 913 
deceased women have been found. As each death certificate is located, a form 
noting the date and cause of death is completed. 

Morbidity Study: A computerized roster of the 1,955 women in the USPHS 
Thyrotoxicosis Follow-Up Study has been compiled. A search of the Massachu- 
setts death certificates found 389 deaths. A questionnaire was mailed to 1,566 
women on April 2, 1979. A second mailing to those who did not respond went out 
May 28, 1979. To date, 81% of the 1,955 women have been found dead or have 
returned a completed questionnaire. Those who have not yet responded are being 
traced through the medical records, telephone books, and Residents' Lists. The 
scheduling and supervision of the clinical examinations have begun. A special 
form to record the results of the physical examination and laboratory tests has 
been designed. So far, 490 patients have been examined. 

Project Officers: Elizabeth P. Anderson, Ph.D., Bruce Nisula, M.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: $78,500 



155 



CONTRACT RESEARCH SUMMARY 

Title: Surgical Adjuvant Chemotherapy in Patients with Breast Cancer 

Principal Investigator: Dr. David L. Ahmann 

Performing Organization: Mayo Foundation 

City and State: Rochester, MN 

Contract Number: NOl-CB-33899 

Starting Date: 6/30/73 Expiration Date: 6/29/81 

Goal: To assess the effects of surgical adjuvant chemotherapy with or without 
radiation therapy in patients with operable but prognostically unfavorable 
breast cancer. 

Approach: Patients with unfavorable breast cancer (node + or unfavorable signs) 
are assigned to treatment programs involving mastectomy followed by adjuvant 
chemotherapy with or without radiation therapy. The three treatment arms are 
a) L-PAM, b) CFP and c) CFP with radiation therapy. Because of the therapeutic 
disadvantage to L-PAM in the premenopausal setting, this treatment arm was 
dropped for this group of patients. The adjuvant chemotherapy was initiated up 
to six weeks in the postoperative period and when radiation is given it is given 
concomitantly with the chemotherapy. Chemotherapy programs are continued for a 
total of ten cycles or upon the appearance of recurrent disease, whichever 
should occur first. 

Progress: The results of these three treatments have been assessed in 293 pa- ^ 
tients, 81 treated with L-PAM, 104 with CFP and 108 with CFP plus radiation 
therapy. Of these patients, 13 (4.4%) withdrew from treatment prior to complet- 
ing the planned ten courses. A total of 211 patients have completed two or more 
years of survival followup. With respect to the premenopausal group of patients, 
L-PAM was found to be distinctly inferior to the utilization of CFP with or with- 
out radiation therapy as a treatment modality expressed both by disease-free in- 
terval and survival. In the postmenopausal group of patients thus far, none of 
the treatments employed have displayed a distinct advantage with respect to dis- 
ease-free interval or survival. Specifically, virtually all patients experienced 
some degree of myelosuppression and no dose response or myelosuppressive effect 
was found to be influential on the disease-free interval or survival with the 
treatment program employed in this study. The dominant site of recurrent disease 
at the time of initial recurrence, however, does show that local and regional 
control was markedly improved with the addition of radiation therapy. Toxicities 
encountered were comparable except for unique toxicities exclusively associated 
with radiation therapy including radiation pneumonitis, esophagitis, pericardi- 
tis, radionecrosis and brachial plexopathy. The major morbidity encountered, 
however, in this group was in one patient necessitating hospitalization for radi- 
ation pneumonitis and two patients undergoing pericardiectomy for pericarditis, 
and one patient with a moderately severe plexopathy. Lymphedema was also ob- 
served about twice as commonly and was about twice as severe in the radiation 
treatment group. 

Project Officer: Mary E. Sears, M.D. ^1 

Program: Breast Cancer Treatment 
FY 81 Funds: 

156 I 



• 



CONTRACT RESEARCH SUMMARY 

Title: NCI Sera Bank Facility for the Breast Cancer Task Force 

Principal Investigator: Dr. Vay Liang W. Go 

Performing Organization: Mayo Foundation 

City and State: Rochester, MN 

Contract Number: NOl-CB-84296 

Starting Date: 9/1/77 Expiration Date: 8/31/82 

Goal: To establish and maintain a storage facility for serum specimens to be 
used by the Breast Cancer Task Force in a program designed to search for bio- 
logical markers in breast cancer. 

Approach: Sera specimens will be secured from breast cancer patients, benign 
disease patients, controls, and a screening population under separate contracts 
setting up a patient resource. The material will be processed, recorded and 
stored in -70 °C deep freezers under easily retrievable conditions with all 
clinical data available. The sera will be used in the search for and verifica- 
tion of new breast cancer markers. 

Progress: Collection and inventory methods have been developed. A special 
vial has been obtained and supplied to the collection areas. An operational 
shipping schedule has been established on a regular basis. Samples have been 
catalogued and systematically stored. Inventory during the forty-fourth month 
of the projects is listed: 

Inventory Status April 1981 No. Patients No. Vials 

Screening Project (Columbia, MO) 11,379 122,080 

Operative Patients (Grand Rapids, MI) 2,978 32,253 

Operative Patients (Wilmington, DE) 575 5,917 

TOTAL: 14,932 160,250 

Collected Since April 1980 No. Patients No. Vials 

Screening Project (Columbia, MO) 1,837 19,119 

Operative Patients (Grand Rapids, MI) 1,077 12,445 

Operative Patients (Wilmington, DE) 93 1,009 

TOTAL: 3,007 32,573 

Since June 4, 1979, 8 coded serum panels have been shipped to investigators 
for evaluation of new breast cancer markers. No. Patients No. Vials 

683 717 



Project Officers: Ihor J. Masnyk, Ph.D., Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: $100,000 



157 



CONTRACT RESEARCH SUMMARY 

Title: Prognostic Factors for Disseminated Cancer in Patients with Early 
Breast Cancer 

Principal Investigator: Dr. Paul Peter Rosen 

Performing Organization: Memorial Hospital 

City and State: New York, NY 

Contract Number: NOl-CB-74097 

Starting Date: 8/1/77 Expiration Date: 7/31/81 

Goal: To determine the frequency of subsequent metastatic breast cancer in a 
group of females with "early" invasive carcinoma and to identify those clinical 
and/or pathologic features or groups of features that are indicators of a high 
or low risk for dissemination of carcinoma. 

Approach: A study population of 250 women with primary infiltrating breast 
carcinomas 1.0-2.0 cm in diameter, and a subset of 100 women with malignant 
tumors 1 cm or less in diameter will be identified in a retrospective study 
(1964-1969) . These patients will have been treated by at least a modified 
radical mastectomy and have been found to have negative lymph nodes. The rela- 
tionships observed between pathologic and clinical data and survival will be 
evaluated. Patients with primary tumors 2 cm or less with positive Ijnnph nodes 
will also be studied to provide a basis for comparing the distribution of vari- 
ous features. 

Progress: A 10-year follow-up study of 382 women with Stage I (T]_NqMq) breast 
carcinoma revealed recurrence and/or death due to cancer in 16%. Among 134 pa- 
tients (35%) with a primary tumor 1.0 cm or less in diameter (Group A), 7% had 
recurrence and 5% died of breast carcinoma. Recurrences were observed in 21% 
of the 248 women with a tumor 1.1 to 2.0 cm in diameter (Group B) and 15% died 
of disease. These differences in recurrence and mortality were statistically 
significant. All recurrences were due to infiltrating duct or lobular carci- 
noma which accounted for 91% of the 382 carcinomas. Most strongly linked to 
recurrence was the finding of tumor emboli in lymphatics of the breast. This 
was found in 23 Group B patients and 10 of them (43%) died of disease. No 
recurrences were observed among the 7 Group A patients with lymphatic emboli. 
Other features associated with a significantly increased risk of recurrence 
were poorly differentiated carcinoma, marked lymphoid reaction to tumor, and 
menarche before age 12 or after age 14. No combination of variables proved to 
identify a subset of patients with an especially increased or low risk of re- 
currence. Stage I patients with lymphatic tumor emboli in the breast surround- 
ing a carcinoma 1.1 to 2.0 cm in diameter have a sufficient risk for recurrence 
to warrant consideration of adjuvant systemic therapy. A very low risk of 
recurrence was observed for the following: any tumor 1.0 cm or smaller; and 
tubular, medullary or colloid carcinoma up to 2.0 cm. Analysis of 142 T]_N2_Mq 
patients is currently under way. 



Project Officers: D. Jane Taylor, Ph.D., Donald E. Henson, M.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



158 



CONTRACT RESEARCH SUMMARY 

Title: Longitudinal Studies of Biologic Markers in Breast Cancer Patients 

Principal Investigator: Dr. Morton K. Schwartz 

Performing Organization: Memorial Hospital 

City and State: New York, NY 

Contract Number: NOl-CB-74206 

Starting Date: 8/1/77 Expiration Date: 7/30/82 

Goal: To assay hximan breast cancer tissue and sera prior to mastectomy and 
throughout the clinical course of the disease for potential markers, to deter- 
mine how effective such markers would be for early detection of breast cancer 
and its recurrence, and to gain an understanding of the relationship between 
the concentration of the marker substances in the individual tumor and in the 
servmi levels of the host. 

Approach: Serxom and tissue specimens from patients entering for diagnosis and 
for primary therapy are assayed for carcinoembryonic antigen (CEA), sialyltrans- 
ferase, phosphohexose isomerase (PHI), spermine, ferritin, lactic dehydrogenase 
(LDH) and its isoenzymes, fl-hCG, y-glutamyltranspeptidase (y-GTP), a-fetoprotein 
and IgA. In tissue alone hormone receptors, total protein, DNA and glucose-6- 
phosphate dehydrogenase and measured and in serum aspartate aminotransferase 
(GOT), alkaline phosphatase, calcium and albumin. Serum specimens are collected 
prior to surgery, before discharge and then sequentially every 3 months the 
first year and every 6 months thereafter. The tissue is a portion of that ob- 
tained for hormone receptor assay at the time of biopsy or mastectomy. Tissue 
and serum biochemical data are compared and changes and differences related to 
the clinical course of the patient as well as demographic and pathology findings. 

Progress: As of December 1, 1980, 393 patients had been entered into the study 
and additional patient accrual was terminated. Collection of serum specimens 
and their assay continues. There have been 56 patients in whom clinical recur- 
rence has been observed. In some of these serum marker elevations preceded the 
clinical observation. However, there were also patients with recurrence in whom 
marker changes were not observed. The statistical technique of logistic regres- 
sion is being used in an attempt to develop a multivariate model to predict 
recurrence from the initial tissue and pre-surgical blood data as well as other 
variables including nodal status and tumor size. 



Project Officer: D. Jane Taylor, Ph.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



159 



Title; 



CONTRACT RESEARCH SUMMARY 

Metabolism of Polycyclic Aromatic Hydrocarbons in the Induction of 
Mammary Tumor 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. A. Monaem El-hawari 
Midwest Research Institute 
Kansas City, MO 



Contract Number; 
Starting Date: 



NOl-CB-84227 
9/1/79 



Expiration Date: 8/31/81 



Goal: To determine whether differences in the disposition, metabolism, and 
macromolecular binding are related to differences in mammary tumor incidence 
caused by polycyclic aromatic hydrocarbons. 

Approach: Carcinogenesis studies are being performed in sexually mature female 
rats (50 day-old) from Sprague-Dawley (SD), Long-Evans (LE) and Wistar-Lewis 
(WL) strains treated with single doses of MC or DMBA. The disposition, metabo- 
lism and DNA binding of these and other related carcinogens were studied in 
female rats of the 3 strains, in male SD rats and in younger (30 day-old), 
older (110 day-old), pregnant and lactating SD rats. 

Progress: After treatment with both carcinogens, WL rats showed the highest 
susceptibility. Tumors were induced readily in SD rats while LE rats were the 
least susceptible. DMBA was more effective than MC as a mammary tumor inducer 
in the 3 strains. Most tumors induced in the WL and SD rats were malignant 
while LE rats developed mainly benign tumors. Disposition studies showed no 
major differences among the 3 strains in the mammary uptake of both carcino- 
gens. A slower rate of clearance was found in the LE rats but the metabolic 
profiles were not different from the other strains. Mammary microsomes metabo- 
lized MC, DMBA and BaP to a variety of oxidative products. Also, freshly iso- 
lated mammary cells were active in converting these compounds to ethyl acetate- 
and water-soluble metabolites which included both K-region and non-K-region 
dihydrodiols. HPLC metabolic profiles from mammary cells differed from those 
of liver cells and subcellular fractions. Also, mammary enzymes catalyzed the 
conversion of the 3 hydrocarbons to metabolites that reacted with exogenous 
DNA. Binding was modified by inducers and inhibitors and was highest to DMBA 
followed by MC and BaP. This correlates with the carcinogenic potency of the 
3 hydrocarbons. MC and DMBA binding to exogenous DNA was highest when cata- 
lyzed by mammary microsomes from 30 day-old rats. This contrasted with the in 
vivo findings in which binding to mammary DNA was highest in the 50 day-old 
animals. 



Project Officer: Chester V. Piczak, B.S. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



^ 



160 



CONTRA.CT RESEARCH SUMMARY 



Title: 



Lipid Levels and Cholesterol Metabolism in Relation to Human Breast 
Cancer Risk 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-84318 
9/30/78 



Dr. Angelos E. Papatestas 
Mount Sinai Medical Center 
New York, NY 

Expiration Date: 9/29/82 



Goal: To evaluate whether dietary or metabolic factors influence breast cancer 
risk by inducing systemic and/or local changes at the target organ affecting 
host's resistance. 

Approach: Matched case-control study. Matching criteria: age, ethnicity, 
menopausal status. Data on past medical and family history, weight, obesity, 
dietary habits, serxmi lipids, fecal steroid metabolites, tissue cholesterol and 
fatty acids and case-control differences in these variables are evaluated. 
Correlation between these parameters and known risk factors, or tumor charac- 
teristics (tumor stage, presence of hormone receptors) are examined. 

Progress: Significant differences between cases and controls were noted in: 



VARIABLES 



# OF 

PAIRS CONTROLS 



MEAN DIE- PAIRED 
CASES FERENCE T-TEST P 



Serum 

Cholesterol (mg/dl) (CHOL) 

CHOL Postmenopausal Women 

Triglycerides (mg/dl) 

Free fatty acids (mEq/L)* 

Fecal Specimens 

Total steroids 

Neutral steroids 

% Cholesterol degraded 

Dietary Intake 

% of calories from MFA** 



140 


193+36 


204+40 


-11+47 


2. 


,71 


<0.01 


49 


202+37 


226+35 


-24+46 


3. 


.71 


<0.001 


135 


9^63 


119+67 


-20+86 


2. 


,71 


<0.01 


74 


.30+.12 


.35+. 15 


-.05+. 17 


2, 


.49 


<0.025 


39 


44+31 


61+36 


-17+47 


2. 


,18 


<0.05 


39 


29+23 


43+29 


-14+37 


2, 


.14 


<0.05 


39 


52+32 


68+21 


-16+39 


2- 


.58 


<0.02 



11 



16+3 



12+4 



+4+6 



2.26 <0.05 



*Premenopausal women 



**Monounsaturated fatty acids 



These differences were present both in matched pairs of cases to controls with 
benign breast disease and pairs of cases to controls without benign breast 
disease. The magnitude of the differences varied in relation to ethnicity and 
menopausal status. The findings that serum lipids and fecal steroid metabo- 
lites are higher in cases compared to controls may partly reflect the observed 
differences in the type of dietary fat intake. Other factors (physical activ- 
ity, psychosocial stress) affecting serum levels need further investigation. 
As tumor load increases did not correlate with lipid levels the observed dif- 
ferences probably precede rather than follow the appearance of breast cancer. 



Project Officers: Elizabeth P. Anderson, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: $80,000 



Kenneth Lippel, Ph.D. 



161 



CONTRACT RESEARCH SUMMARY 



Title: Interrelationships Among Diet, Steroid Hormone Metabolism, and Human 
Breast Cancer 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number; 
Starting Date: 



NOl-CB-84229 
9/30/78 



Dr. Marvin A. Kirschner 

Newark Beth Israel Medical Center 

Newark, NJ 



Expiration Date: 9/29/81 



Goal: To investigate steroid hormone production rates and metabolism in rela- 
tion to obesity and dietary patterns. 

Approach: This will be an in-depth study of androgen, estrogen and Cortisol 
production, metabolism, and excretion in a cohort of 40 obese women undergoing 
drastic weight reduction and diet manipulation. The women will be studied 
prior to weight loss, after weight loss and stabilization on a high protein, 
low fat diet, and after isocaloric exchange to a more typical high carbohydrate, 
high fat diet. Values will be compared under three sets of conditions and fur- 
ther compared with those in lean, age-matched controls. 

Progress: Baseline studies have been performed in 54 obese and 32 normal women, 
of which 34 and 22, respectively, have been worked up. 



Obese 



Normal Controls 



p Value 



MCR Androstenedione (A) 

Plasma 

\ Production Rate 

ConversionA— >E]^ 

Conversion^— 'Testosterone 

Urinary E^ Prod. Rate 



2140 


<0.001 


121 


N.S. 


2.38 


<0.05 


1.51% 


<0.05 


12.14% 


<0.05 


98 


<0.05 



3143 L/day 
110 ng/dl 
3.53 mg/day 
2.17% 
7.59% 
139.0 ^g/day 

These data demonstrate increased clearance and production rates of A, as well as 
altered metabolism of A~ to both testosterone and E^. Urinary E^ production 
rates are elevated in obese women, due largely to increased extragonadal metabo- 
lism of A to E]^. To date, 32 normal volunteers have been studied during normal 
diet, high protein diet and high carbohydrate diet. Analyses of basal dietary 
patterns (previous week recall) showed no major differences in control vs obese 
women, except in total caloric consumption. Hormonal studies reveal no signifi- 
cant differences in A and E^ production, and peripheral metabolism on these 
differing diets. Our major problems continue to be a) high patient drop-out 
prior to weight loss to ideal weight, and b) instability of weight once basal 
levels are reached, requiring longer periods of stabilization prior to repeat 
studies. 



Project Officers: Elizabeth P. Anderson, Ph.D., Lynn Loriaux, M.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



16 2 



i 

i 
' i 



CONTRACT RESEARCH SUMMARY 

Title: Relation of Intestinal Flora to Breast Cancer in Humans 

Principal Investigator: Dr. Sherwood L. Gorbach 

Performing Organization: New England Medical Center 

City and State: Boston, MA 

Contract Number: NOl-CB-74104 

Starting Date: 7/1/77 Expiration Date: 12/31/80 

Goal: To determine the influence of dietary factors on the metabolism of 
estrogens by bacteria in the large bowel. 

Approach: The estrogen profiles are determined in the serum, urine, and feces 
of young and older women eating "Western" or vegetarian diets and in women with 
a history of breast cancer. In addition, serum SHBG and prolactin are being 
determined. Novel fecal metabolic transformation of estrogens and the possible 
role of these transformations in the etiology of breast cancer are being inves- 
tigated. 

Progress: The total of 54 subjects in 5 groups have completed four 72-hour 
collections of urine, feces, and dietary records plus 3 daily blood samples 
within a one-year period. The laboratory analysis of all the samples has been 
completed and statistical analysis is now being completed. The main conditions 
being analyzed are ovarian state (young menstruating women versus postmeno- 
pausal women) and diet (vegetarian versus omnivores). A fifth group of post- 
menopausal subjects with breast cancer has also been studied. Analysis of the 
dietary data indicates that the young omnivores consume 41.1% of their calories 
as fat and the old vegetarians 30.2% of their calories as fat. Protein intake 
is similar but dietary fiber is higher in the vegetarian, approximately 30 g a 
day versus 12-18 g a day in the omnivores. Analysis of the dietary records 
includes 27 separate nutrients with comparison of means as well as the analysis 
of variation within each group. The fecal data indicate that vegetarians have 
almost a two-fold increase in wet and dry fecal weight. Sex hormone binding 
globulin was analyzed in the seriim and showed that the breast cancer group had 
half the level of the other four groups. In contrast, morning and mid-day 
serum prolactin determinations showed no difference between groups. The data 
on estrogen levels in plasma, urine, and feces are in the process of being ana- 
lyzed. Fecal excretion of conjugated estrogens is increased 2-3-fold in vege- 
tarians and the urinary excretion of estriol-3-glucuronide is higher in omni- 
vores. This result further supports the other findings that omnivores absorb 
more estrogens from the intestinal tract. 



Project Officers: Elizabeth P. Anderson, Ph.D., Carl Smith, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



163 



Title; 



CONTRACT RESEARCH SUMMARY 

Produce and Identify Antibodies to Collagens/Procollagens and/or 
Related Enzymes 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Burton Goldberg 

New York University Medical Center 

New York, NY 



Contract Number; 
Starting Date: 



NOl-CB-84314 
9/1/78 



Expiration Date: 8/31/81 



Goal: To produce specific antibodies to the carboxyterminal nonhelical 
sequences of human type I procollagen in sufficient quantities for storage and 
distribution to other investigators. 

Approach: Culture human skin fibroblasts (CRL1121) in roller bottles and iso- 
late the procollagen in the culture medium, then subject it to specific bacte- 
rial collagenase digestion. Purify the carboxyterminal fragment by ion exchange 
chromatography, then produce antibodies in appropriate animal species (rabbit, 
goat, sheep). Purify the antisera by absorption techniques and/or affinity 
chromatography and confirm their specificity by Ouchterlony technique, Immuno- 
electrophoresis, passive hemagglutination, and radioimmunoassay. 

Progress: To date 120 liters of medium from confluent cultures of human skin 
fibroblasts were obtained. Approximately 1 gram of procollagen-enriched lyophi- 
late was harvested and a portion was subjected to collagenase digestion to gene- 
rate the non-helical carboxyterminal propeptide fragment from procollagen. The 
propeptide fragment was purified by ion exchange chromatography on DEAE-cellu- 
lose and 50 mg of the pure protein was prepared. One sheep and 7 rabbits were 
immunized with the preparation. Some rabbit antisera have titers as high as 
1/80,000 and sheep antisera have titers 1/10,000 to 1/14,000 by radioimmunoas- 
say. A new immunization protocol raised the titers to 1/40,000. Affinity puri- 
fied IgG (3.5 mg) was recovered from 30 ml of rabbit antiserum which had an 
original titer of 1/60,000. The purified IgG reacts specifically with the pro- 
peptide antigen and lacks cross-reactivity for native human collagen. IgG gives 
strong and specific indirect immunofluorescence with cultured human fibroblasts 
when used at a concentration of 2.8 /ig/ml. 

Presently, the stocks of affinity-purified IgG are as follows: 25 ml of 
rabbit IgG at a concentration of 142 ^lg/ml in PBS and 10 ml of sheep IgG at a 
concentration of 142 A^g/™! in PBS. In the current year plans are to at least 
double the amounts of affinity-purified antibodies on hand and to run additional 
tests of their specificity for the carboxyterminal propeptides of type I human 
procollagen. 



Project Officer: Chester V. Piczak, B.S. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



< 



164 



CONTRACT RESEARCH SUMMARY 

Title: Epidemiology of Minimal Breast Cancer in Initially Asymptomatic Women 

Principal Investigator: Dr. Bernard S. Pasternack 

Performing Organization: New York University Medical Center 

City and State: New York, NY 

Contract Number: NOl-CB-74103 

Starting Date: 7/15/77 Expiration Date: 6/30/81 

Goal: Investigate by means of risk factor analysis whether minimal breast can- 
cer in initially asymptomatic women is essentially different from clinical 
breast cancer or whether it is an early manifestation of the same disease; 
identify risk factors associated with tumor aggressiveness. 

Approach: We have obtained and will analyze demographic and medical data re- 
corded at the Guttman Breast Diagnostic Institute, which since 1968 has been 
offering free breast examinations to women living in New York City. The study 
group will consist of an estimated 874 valid cases out of a total of 1,386 
detected through November 21, 1979, and approximately 2,168 valid controls out 
of a random selection of 2,700. After exclusions due to symptomaticity, 478 
cases and 1,712 controls will remain. Two independent histopathologic diagno- 
ses are being performed; one at New York University Medical Center (NYU) and 
the other at Saint Barnabas Medical Center (SB). Cases will be compared 
according to three tumor criteria: a) level of invasiveness, b) level of 
aggressiveness, and c) histologic type. Comparison of all cases to controls 
should result in estimates of relative risk within the range found at other 
screening centers, whereas the comparison of clinical cases to controls should 
provide estimates within the same order of magnitude as registry data. 

Progress: Protocols have been established for all phases of the study: col- 
lection of demographic and screening data, pathology review, and mammography 
review. Basic identification forms have been coded, keypunched, and filed for 
all 1,386 cases, and 2,594 controls. Examination code forms for about 3,980 
participants have been coded, keypunched, and filed. Initial and recall his- 
tories have been coded, keypunched, and filed for about 3,850 participants. 
Pathology material has been requested for 813 cases and received for 625 
(76.9%). Material for 58 cases is definitely unavailable, due to closed hos- 
pitals and lost slides, and for 45 cases will not be requested due to prior 
mastectomy. Six hundred fifty-eight cases remain outstanding. Mammography 
review has been performed for 417 cases and 87 controls, the NYU pathology 
review for 538 cases, and the SB pathology review as well for 529 cases. Of 
those for which two reviews have been completed, the initial rate of consensus 
is 69.2%. Consensus review has been able so far to resolve all remaining dif- 
ferences. Identification of inconsistencies in the data by means of specif- 
ically designed computer programs is being performed for the 19,540 data 
records computerized thus far. These inconsistencies are being resolved by 
examination of participants' files at both NYU and the Guttman Institute. 

Project Officers: Elizabeth P. Anderson, Ph.D., Benjamin Hankey, Sc.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



165 



CONTRACT RESEARCH SUMMARY 

Title: Development of an Assay for Genetic Damage to Mammary Gland Cells 

Principal Investigator: Dr. Steven D'Ambrosio 

Performing Organization: Ohio State University 

City and State: Columbus, OH 

Contract Number: NOl-CB-84226 

Starting Date: 9/1/78 Expiration Date: 8/31/81 

Goal: To develop an inexpensive, rapid, and quantitative assay for detecting 
genetic damage and its repair in mammary cells in vivo following exposure to 
carcinogenic agents. 

Approach: Measure DNA damage directly by an assay using S]^ endonuclease. 
Determine changes in the size of nonradiolabeled DNA resulting from endonucle- 
ase treatment using alkaline or neutral sucrose gradient sedimentation. After 
sedimentation, the nonradiolabeled DNA will be quantitated by a spectrof luoro- 
metric technique. Quantitate the mammary epithelial cell DNA damage by double 
label autoradiography. DMBA sensitive and resistant rat strains and noncarci- 
nogenic and less carcinogenic analogs, 2-FL-DMBA and 5-FL-DMBA, will be used 
for these studies. 

Progress: Since the inception of the contract this laboratory has: a) devel- 
oped techniques for the isolation of mammary nuclei essentially free of conta- 
minating lipids; b) developed a technique that permits sedimentation and detec- 
tion of nanogram quantities of nonradiolabeled mammary DNA; c) quantitated the 
molecular weight of such DNA (determination of the number of breaks induced); 
d) synthesized DMBA and its fluorinated analogs and ENU; e) developed methods 
for separation and purification of these cold and radiolabeled compounds; f) 
quantitated the number of DNA-carcinogen adducts produced in mammary gland and 
compared it to that produced in other tissues; g) determined the level of uri- 
nary metabolites produced by radioactive and nonradioactive DMBA and its fluo- 
rinated analogs and h) begun to correlate the types and quantities of damage 
to the carcinogenic potential of the compounds tested. 

These studies have led to the development of an inexpensive, rapid and 
quantitative assay for detecting in vivo genetic damage of mammary gland cells. 
Further experiments are in progress and should be completed during year three 
of this contract to determine not only the amount of DNA damage induced but 
also to correlate the rate of removal with the tumor susceptibility of carcino- 
gen and noncarcinogen compounds tested as a function of both rat strain, dose, 
age and location of mammary gland. 



< 



Project Officer: Chester V. Piczak, B.S. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



166 



CONTRACT RESEARCH SUMMARY 

Title: Suppression of Endocrine Function by Systemic Agents for Breast Cancer 
Therapy 

Principal Investigator: Dr. Richard J. Santen 

Performing Organization: Pennsylvania State University 

City and State: Hershey, PA 

Contract Number: NOl-CB-53851 

Starting Date: 5/15/75 Expiration Date: 5/14/82 

Goal: To produce suppression of endocrine function with aminoglutethimide (AG), 
to establish the mechanism of steroid inhibition, and to compare the effects of 
surgical adrenalectomy, antiestrogen therapy, and aminoglutethimide in human 
breast carcinoma. 

Approach: Female patients with inoperable, recurrent, or metastatic breast car- 
cinoma and whose tumors are either estrogen receptor positive or unknown are 
selected for study. Women are randomized into two separate therapeutic trials: 
a comparison of medical adrenalectomy (AG + hydrocortisone) vs. surgical adrena- 
lectomy, and of AG vs. the antiestrogen, tamoxifen. Extensive endocrine studies 
evaluate the effects of AG on extra-adrenal estrogen production, androgen, pro- 
gestin, and prolactin secretion, and steroid metabolism. 

Progress: Women with metastatic breast cancer are entered into protocols uti- 
lizing surgical adx, medical therapy with aminoglutethimide (AG) and tamoxifen 
(Tam). In a randomized trial of 96 patients, a 53% objective response rate to 
AG plus hydrocortisone (HC) and a 43% regression rate to surgical adx were ob- 
served. Medical treatment with AG and surgical adx produced a similar reduction 
of estrogen levels, whereas androgen secretion was preserved in the medical 
group. Four women treated initially with surgical adx experienced additional 
estrogen suppression upon addition of AG and in 2, partial objective tumor 
regression resulted. Systemic studies of adrenal reserve immediately after 
stopping AG revealed normalization of basal Cortisol levels and return of stress 
responses within 72 hr. A randomized trial of AG-HC vs. the antiestrogen, 
tamoxifen, entered 61 patients. Forty-eight percent responded objectively to 
AG-HC and 42% to Tam (p = NS). However, bone lesions appeared more favorable 
to AG therapy (7/13 CR + PR = 54%) than to Tam (4/18 CR + PR = 22%). Cross-over 
responses to Tam in AG-HC -resistant patients occur infrequently but Tam-resis- 
tant women appear to have secondary responses to AG-HC. Hormonal levels in 147 
AG-treated patients revealed equal estrogen suppression in objective responders 
as in patients with progressive disease but DHEA-S and androstenedione levels 
were higher during treatment in patients with progressive disease. Overall, 
these studies suggest that AG-HC is a logical alternative to surgical adx and 
produces clinical responses which probably differ from those induced by Tam. 



Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: $72,000 



167 



CONTRACT RESEARCH SUMMARY 

Title: Prognostic Significance in Breast Cancer of Regional Lymph Node Immune 
Response 

Principal Investigator: Dr. Susanna Cunningham-Rundles 

Performing Organization: Sloan-Kettering Institute 

City and State: New York, NY 

Contract Number: NOl-CB-84228 

Starting Date: 9/1/78 Expiration Date: 8/31/81 

Goal: Evaluation of the relevance of regional lymph node immune response to 
prognosis in breast cancer. This will require an assessment of cell-mediated 
immune reactivity in well-standardized in vitro assays to defined antigens 
associated with the development of disease. 

Approach: Immunological studies will be performed on Ijnnph node cells of 100 
patients with primary operable breast cancer, using a total of 4-6 lymph nodes 
per patient including both negative and positive nodes, ideally from each level 
(I-III). In addition, control lymph nodes will be obtained from 50 other pa- 
tients. The assays to be done on both peripheral blood and lymph node mononu- 
clear cells include leukocyte migration Inhibition factor (LMIF) and lymphocyte 
transformation tests, the latter comprising PHA, Candida, E^. coli , T-antlgen 
and MuMTV. A third class of newer tests includes natural killer cells, sup- 
pressor cells, cell surface markers and monocyte function assays. Clinical and 
demographic data as well as histopathologic data on tumor size and histologic 
type will be obtained. Three-year follow-up information on recurrence and sur- 
vival status will be recorded. Correlation of all factors will be statis- 
tically analyzed. 

Progress: Principal findings are as follows: 30% of patients with breast can- 
cer had reduced or negative NK activity of PBL toward K562 target cells. About 
40% of patients with tumor positive nodes and 25% of patients with tumor nega- 
tive nodes had negative PBL NK activity. Regional lymph node cells (RLNC) from 
21% of patients had significant NK activity toward K562 in contrast to control 
RLNC including RLNC from patients with benign lesions where positive NK activ- 
ity was not observed. Increased numbers of Ty lymphocytes were found in 70% 
of patients' RLNC and 40% of PBL; these increases did not have a simple corre- 
lation with NK activity. Neither PBL nor RLNC were found to have suppressor 
cell activity in either autologous or allogeneic MLR. RLNC were markedly 
stronger as stimulating cells in MLR proliferative responses to mitogens, anti- 
gens and putative disease-related antigens were different in patients with tu- 
mor positive and negative nodes, suggesting tumor influence on RLNC activation. 



( 



Project Officers: D. Jane Taylor, Ph.D., Donald E. Henson, M.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



168 



CONTRACT RESEARCH SUMMARY 

Title: Epidemiologic Characteristics of Medullary and Lobular Breast Cancer 

Principal Investigator: Dr. Paul P. Rosen 

Performing Organization: Sloan-Kettering Institute 

City and State: New York, NY 

Contract Number: NOl-CB-63997 

Starting Date: 6/30/76 Expiration Date: 12/29/80 

Goal: To identify and describe morphologic features of human female mammary 
carcinoma which are significantly related to known risk factors for the devel- 
opment of such carcinomas. 

Approach: Detailed epidemiologic information was obtained by interviews from 
women with mammary carcinoma and correlated with pathologic features of the 
tumors. The analysis of the data emphasized comparison of medullary and lobu- 
lar carcinoma with duct carcinoma. 

Progress: A total of 1,227 patients have been accessioned into the study. 
Among the cases reviewed, 81 women were found to have only medullary carcinoma, 
38 only in situ lobular carcinoma, and 59 only infiltrating lobular carcinoma. 
In addition, 42 patients had lobular carcinoma (38 invasive, 4 noninvasive) 
combined with another type of carcinoma, and 11 patients had medullary carci- 
noma associated with another type of carcinoma. Interviews have been conducted 
with 98% of the patients. 

Patients with medullary carcinoma had a significantly lower mean age at 
diagnosis than those with duct and lobular carcinoma. There was a dispropor- 
tionately high number of non-whites in the medullary carcinoma group. A sec- 
ondary correlation with medullary carcinoma was a higher frequency of birth in 
the southern part of the United States as well as in South and Central America. 
The number of Oriental women was too small for analysis. Patients with medul- 
lary carcinoma tended to weigh more at diagnosis than those with other types 
of carcinoma, but the differences were not significant when evaluated in terms 
of age and optimum weight. No correlation between height or height-weight 
indices and tumor type was observed. Preliminary analysis of methods of clin- 
ical examination indicates significant relationships between the frequency of 
physician and mammographic examination and tumor stage. No correlation between 
frequency of self-examination and tumor histology has been found. Overall, 32% 
of patients had at least one female relative treated for breast cancer, ranging 
from 29% for patients with infiltrating duct carcinoma to 42% for those with 
lobular carcinoma in^ situ . There was a strong association of medullary carci- 
noma with positive maternal history of breast cancer while lobular carcinoma 
was associated with sister pedigrees. 



Project Officers: Elizabeth P. Anderson, Ph.D., Roger Connelly 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



169 



CONTRACT RESEARCH SUMMARY 
Title: Biochemical Analysis of Human Breast Cyst Fluid (BCF) 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number; 
Starting Date: 



NOl-CB-53853 
6/30/75 



Dr. Morton K. Schwartz 
Sloan-Kettering Institute 
New York, NY 



Expiration Date: 9/14/82 



Goal: To establish the biochemical and immunochemical composition of human 
BCF; to search for a marker which might be useful in understanding the forma- 
tion of cysts, their treatment, and the risk of patients with cysts developing 
cancer. 

Approach: BCF from patients with cystic mastopathy will be analyzed for poly- 
peptide and steroid hormones, tumor-associated antigens, enzymes, lipids, min- 
erals, and trace elements. These data will be correlated with each other and 
with the demographic information. Fluid will be analyzed from patients with 
recurrent cysts and clinical follow-up will be obtained on all patients. 

Progress: Patient entry has been completed. There have been 1,044 patients 
(1,830 specimens) entered into the study. About 25% of the patients have had 
recurrent cysts and additional specimens submitted for analysis since their 
initial aspiration. As previously noted, the concentration in many BCF speci- 
mens of CEA, a-fetoprotein, fl-subunit hCG, calcium, copper, zinc, total protein,^ 
albumin, cholesterol, phosphohexose isomerase, y-glutamyltranspeptidase, B-glu- 
curonidase, lactic dehydrogenase, amylase, dehydroisoandrosterone, androsterone, 
and their sulfates are many-fold the concentrations of these constituents found 
in serum. Correlation of BCF concentrations for many of these constituents in 
the same patients was found to be significant. The specimen variation was much 
less in specimens from the same patients obtained either at the same time or on 
subsequent aspiration than that from patient to patient. Notwithstanding the 
large BCF to serum gradient, isotope studies have indicated a flow influx and 
efflux of the steroids into the cyst fluid. The fluid appears to contain mate- 
rial both of a secretory nature and that resulting from cell breakdown. BCF 
contains low but detectable levels of terminal deoxynucleotidyl transferase 
(TdT) and DNA polymerases were observed in BCF and in supernatant from cells 
from BCF maintained in culture media. Preliminary biostatistical analysis has 
indicated that in the 40-65 years age group where the cyst occurrence is most 
prominent, the incidence of breast cancer in cyst patients is 625 per 100,000 
as compared to 144 per 100,000 from estimates in normal women. Only follow-up 
of patients is continuing to determine the true incidence of breast cancer in 
cyst patients and to establish whether this incidence is related to the biochem- 
ical findings. 



Project Officers: Bernice T. Radovich, Ph.D., R. Quentin Blackwell, Ph.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



170 



ii 



CONTRACT RESEARCH SUMMARY 

Title: Growth and Passage of Primary Culture of Normal Mammary Epithelial 
Cells 

Principal Investigator: Dr. Dayton S. Misfeldt 

Performing Organization: Leland Stanford Jr. University 

City and State: Stanford, CA 

Contract Number: NOl-CB-74094 

Starting Date: 7/1/77 Expiration Date: 6/30/81 

Goal: To define conditions for the growth and passage of normal mammary epi- 
thelial cells in culture. 

Approach: Conditions for growth and passage are determined by colony and cell 
count with an established mouse mammary cell line (NMUMG) and primary cultures 
of BALB/c mouse mammary gland. 

Progress: Conditions for growth and passage of mammary epithelial cells must 
be considered in the context of a mixed cell population obtained during mammary 
gland dissociation. Differential stimulation of epithelial growth and fibro- 
blast suppression has been investigated: 1) The addition of cholera toxin 
results in a 50% inhibition of fibroblast growth without suppression of epithe- 
lial growth. 2) Serum-derived plasma which attempts to remove the platelet- 
derived growth factor, a powerful stimulus for fibroblast growth, inhibits 
fibroblast growth to a greater extent than it inhibits epithelial growth. 3) 
Collagen as a culture substratum limits fibroblast overgrowth of the epithelial 
cell cultures. Eight epithelioid appearing clones have been isolated and pas- 
saged from primary mouse mammary tissue. However, chromosomal analysis of 
three has shown them to be aneuploid. The use of kidney epithelial feeder 
layers did not increase cloning efficiency of the Namru mammary cell line, so 
rat mammary epithelial cells are currently being tested as feeder cultures. 
If they increase Namru cloning, they will be used to obtain clones of primary 
mouse epithelial cells. 



Project Officers: Chester V. Piczak, B.S., D. Jane Taylor, Ph.D. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



171 



CONTRACT RESEARCH SUMMARY 

Title: Longitudinal Studies of Biologic Markers in Breast Cancer Patients * 

Principal Investigator: Dr. Howard H. Sussman 

Performing Organization: Stanford University School of Medicine 

City and State: Stanford, CA 

Contract Number: NOl-CB-74086 

Starting Date: 8/1/77 Expiration Date: 7/30/82 

Goal: To assay human breast cancer tissue and sera, prior to mastectomy and 
throughout the clinical course of the disease, for potential markers to deter- 
mine how effective such markers would be for early detection of breast cancer 
and its recurrence, and to gain an understanding of the relationship between the 
concentration of the marker substances in the individual tumor and in the serum 
levels of the host. 

Approach: Tumor and sera of patients presenting initially for diagnosis and 
primary therapy (Category 1) will be assayed for the fetoplacental proteins, 
i.e., placental alkaline phosphatase (PAP), human chorionic gonadotropin (hCG) 
a and fl, and carcinoembryonic antigen (CEA); and tests on sera will be repeated 
sequentially every three months. Patients developing recurrences (Categories 2 
and 3) will be Included with a study population from the Northern California 
Oncology Group. Another objective of the study will be to identify new ectop- 
ically synthesized placental membrane proteins, which may be common to both tro- 
phoblasts and neoplastic breast tumor cells. ^, 

Progress: Serum assays have been continued on the 125 patients participating 
in the study. One patient from Category 1 has relapsed into Category 2. Three 
of her four marker levels increased, with two of these becoming abnormal, in the 
period of three to eight months prior to the diagnosis. The patient has since 
declined to be followed. A receptor specific for transferrin has been found in 
neoplastic breast ttmiors. Microsomes from these txmiors demonstrate 11-35% bind- 
ing of transferrin, while those from normal breast tissue show only 2-3% binding. 
A radioimmunoassay has been set up to detect the level of transferrin receptor 
on cell membranes. Preliminary results, expressed as ^g receptor/mg protein, 
show normal breast (0.2) and a benign male breast (0.4) to have low levels, 
while placenta (33.6) has a high level. Eleven breast cancer tumors have been 
assayed and those levels fall between these two limits. Data on the 400 sub- 
jects from whom we have at least one blood sample, including controls, is con- 
tinuing to be collected and entered into a computer for future evaluation. 



Project Officers: D. Jane Taylor, Ph.D., Donald E. Henson, M.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: $27,000 



172 



CONTRACT RESEARCH SUMMARY 

Title: Morphological Properties of Normal and Abnormal Human and Rodent 
Mammary Tissue 

Principal Investigator: Dr. Elinor Spring-Mills 

Performing Organization: State University of New York 

City and State: Syracuse, NY 

Contract Nvunber: NOl-CB-84239 

Starting Date: 8/14/78 Expiration Date: 8/13/81 

Goal: To evaluate structural differences among normal, dysplastic, and cancer- 
ous mammary glands. 

Approach: To describe, quantitate, and compare certain morphological and func- 
tional characteristics of normal, hyperplastic, and carcinomatous human and 
mouse mammary glands in vivo and in vitro . Human tissues obtained at biopsy, 
mastectomy and reduction mammoplasty will be prepared for light microscopy 
(LM), transmission electron microscopy (TEM), scan electron microscopy (SEM), 
and quantitative electron microscopy (QEM) or morphometry. Animal tissue from 
nonpregnant (pubertal and adult), pregnant, and multiparous C3H mice will be 
studied. 

Progress: During the past year, tumors and proliferative lesions from 40 women 
have been explanted and cultured. Survival has been poor to excellent in syn- 
thetic medium for 6-42 days. The effects of no hormones, insulin, hydrocorti- 
sone, ^prolactin, glutamine and vitamin A, alone and in combination, have been 
tested and evaluated at the light microscopy level. A large computer program 
is being written to analyze the interrelationships between the responses of the 
patient's tissues to the organ culture conditions, the pathological diagnosis 
of the breast lesion, the patient's zero time serum hormone levels for prolac- 
tin, progesterone, estrone and estradiol, and other risk factors. This program 
is to generate a model for testing the relationship of particular variables to 
the onset and subsequent course of certain breast diseases and the value of the 
culture systems for assessing tissue responsiveness to hormones, tissue poten- 
tial for proliferation and growth, etc. A pilot immunocytochemical study of 
immunoreactive insulin in human breast tumors is under way. 

The mouse project has been expanded because they found that morphologic 
alterations related to HAN and other lesions of undetermined biologic signifi- 
cance are appearing in the 2 1/2-3 month old C3H/HeJ mice. The breasts, ova- 
ries, uteri, adrenals and oviducts from C3H/HeJ mice have been studied with the 
LM, SEM, and TEM. To better evaluate the data, parallel studies have been 
under way since April 1980 on C3HeB/FeJ mice which have low mammary tumor 
incidence. 



Project Officer: Chester V. Piczak, B.S. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



17; 



CONTRACT RESEARCH SUMMARY 
Title: Purification of Estrogen Receptor in Tangible Amounts 



Principal Investigator: 
Performing Organization: 
City and Country: 

Contract Number: NOl-CB-64074 
Starting Date: 9/30/76 

Goal: To isolate and purify the 
milligram quantities. 



Dr. Giovanni Alfredo Puca 
Universita di Napoli 
Napoli, Italy 



Expiration Date: 5/29/81 
"native" form of the estrogen receptor in 



Approach: Carry out large-scale tissue disintegration and centrifugation for 
isolation of estrogen receptor from calves' uteri. Purify the "native" form of 
estradiol receptor by a combination of affinity chromatography on new adsorbents 
and conventional separation methods. 

Progress: The procedure for the purification of the native form of estradiol 
receptor of calf uterus and recently published (Journal of Steroid Biochemistry, 
12, 105, 1980) was utilized with one modification to render the purification 
feasible when large volumes of cellular extract have to be handled. The tissue 
homogenate is now centrifuged in a refrigerated centrifuge because the capacity 
is much larger than the ultracentrifuges. The low speed supernatant is then 
incubated batchwise with heparin-agarose. After elution of the estradiol bind- 
ing activity by heparin, the protein solution is then subjected to high speed 
centrifugation in an ultracentrifuge. Since the volume of this eluate is usu- 
ally reduced to one tenth the volume of the original cellular extract, the 
ultracentrifugation step becomes much easier. 

The immobilization of receptor on the estradiol containing derivative is 
carried out batchwise to shorten the time of incubation and to avoid loss of 
estradiol binding activity. The estradiol derivative is then packed in a column 
and washed very extensively with various cycles of low and high salt (2 M KCl) 
buffers. The receptor is eluted from the column in a very sharp peak with a 
buffer that contains, in addition to estradiol and chaotropic salt, 10% of 
dimethyl formamide. Sodium dodecyl sulfate polyacrylamide gel electrophoresis 
showed that, after these two simple affinity chromatography steps, receptor was 
at least 70% pure. Depending on the amount of the binding activity originally 
present in the extract, in the last three preparations 60, 96, and 380 /ig of 
pure receptor starting from 450 gm of calf uteri was obtained. 






Project Officers: Chester V. Piczak, B.S., D. Jane Taylor, Ph.D. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



4 



174 



CONTRACT RESEARCH SUMMARY 

Title: Correlations Between Morphology and Epidemiology of Breast Cancer 

Principal Investigator: Dr. Bjdrn Stenkvist 

Performing Organization: University Hospital, Uppsala 

City and Country: Uppsala, Sweden 

Contract Number: NOl-CB-53968 

Starting Date: 6/30/75 Expiration Date: 6/30/81 

Goal: To ascertain possible correlations between morphologic variables such as 
cytologic and histopathologic classifications and established epidemiological 
risk factors of breast cancer. 

Approach: Within a clearly defined area and population (4 Swedish counties) 181 
breast cancer cases occurred during a defined study period (5 months). From 179 
of the patients both epidemiological and morphological data could be obtained. 
From 179 exactly age-matched controls epidemiological data were recorded in the 
same way. The tumors were characterized by computerized cytometry of the cell 
population in the tumors as well as by conventional morphology, resulting in an 
objective atypia measure. 

Progress: In collaboration with Dr. Alan Morrison, Harvard School of Public 
Health, files of the epidemiologic and morphologic data have been established 
on the DEC-20 computer at the Sidney Farber Cancer Institute. Programs have 
been written to select subsets of this information for specific analyses.- Pre- 
liminary analysis has been done to verify the programming and confirm the feasi- 
bility of the computational procedures. These analyses have involved both com- 
parisons of suspected risk factors between cases and controls and comparisons 
of risk factors among groups of cases defined according to histologic character- 
istics. Detailed epidemiologic analysis of the data is beginning. 

The patients have been followed up for the first four years postmastectomy . 
A "malignancy grade" for breast carcinomas has been developed, based on image 
analysis and pattern recognition. This measure of malignancy grade had a high 
prognostic significance and was applicable as an index of recurrence risk in 
each individual case. 



Project Officer: Elizabeth P. Anderson, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



175 



CONTRACT RESEARCH SUMMARY 

Title: Biologic Characterization of "Premalignant" Human Mammary Epithelial 
Hyperplasias 

Principal Investigator: Dr. Hanne M. Jensen, M.D. 

Performing Organization: University of California 

City and State: Davis, CA 

Contract Number: NOl-CB-84316 

Starting Date: 9/18/78 Expiration Date: 9/17/81 

Goal: To establish if angiogenesis is a reliable criterion for precancer of 
the human breast and to characterize by biochemical and immunological markers 
the type(s) of lesions that possess angiogenic capacity from those that do not. 

Approach: Fresh, parenchymal structures and hyperplastic lesions stained with 
methylene blue chloride are isolated and studied for angiogenic potential by 
transplantation onto the iris of female rabbits. Six micron thick frozen sec- 
tions of parts of isolated structures and lesions are stained with fluorescent 
labeled anti-human immunoglobulins G, A and M, to characterize them immunolog- 
ically and with fluorescent labeled estrogen and progesterone to detect hormone 
receptors. 

Progress: From 91 breast specimens 1,077 transplants derived from 784 struc- 
tures and lesions were studied; 200 transplants became nonexperiments because 
they contained no epithelium or floated off the iris. Statistical evaluation 
indicated that 5 successful transplants from 5 different lesions from a given 
case were needed to prevent false negative data for angiogenesis. There were 
409 successful transplants of normal and atypical lobules; 54 of 362 (15%) nor- 
mal lobules were angiogenic. The frequency of angiogenesis was equal in cancer 
associated and noncancerous cases. Four of 23 (17%) atypical lobules from non- 
cancerous biopsies were angiogenic. Fourteen of 23 (61%) atypical lobules from 
cancer associated breasts were angiogenic. The probability of the observed 
differences in angiogenic potential of atypical lobules occurring by chance is 
less than 0.01. A higher proportion of lobules from cancer cases were atypical 
[23 of 95 (24%)] than those from benign cases [23 of 314 (7%)]; chance proba- 
bility is less than 0.0001. Atypical lobules were derived from 10 women with 
cancer. Nine of these had one or more angiogenic atypical lobules. Atypical 
lobules were obtained from eight women without cancer. Four of these had one 
or more atypical lobules that were angiogenic. The probability of this occur- 
ring by chance is greater than 0.05 but less than 0.1. Presumably these four 
women have greater risk for developing cancer at a later date. Most angiogenic 
atypical lobules secrete immunoglobulins and those found in women after age 60 
appear strongly positive for estrogen receptors (ER) if indeed the fluorescent 
estrogen is specific. It is hypothesized that postmenopausally all hormone 
dependent tissues and lesions with low ER content regress due to lack of stimu- 
lation but those with high ER content persist. 



Project Officers: D. Jane Taylor, Ph.D., Donald E. Henson, M.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



176 



CONTRACT RESEARCH SUMMARY 

Title: Growth and Passage of Primary Cultures of Normal Mammary Epithelial 
Cells 

Principal Investigator: Dr. Hideo Masui 

Performing Organization: University of California 

City and State: La Jolla, CA 

Contract Number: NOl-CB-74188 

Starting Date: 8/1/77 Expiration Date: 7/31/81 

Goal: To isolate and maintain growth of pure normal mammary epithelial cells 
in culture. 

Approach: Determine the hormonal and nutritional requirements of established 
animal and human mammary tumor cell lines. Proceed with similar studies on 
primary cultures of human mammary tumors obtained from autopsies and mastecto- 
mies. Determine the hormonal content, blood serum components, and growth fac- 
tors (ovarian, nerve, epidermal, etc.) to prepare a defined media to maintain 
growth and passage of normal mammary epithelial cell lines. 

Progress: Normal human mammary epithelial cells were supplied by Dr. Martha 
Stampfer which were obtained from reduction mammoplasty. The mammary tissue 
was treated with a digestion mixture of collagenase and hyaluronidase to 
release organoid structure-containing epithelial cells and myoepithelial cells. 
The organoids were collected by filtration and centrifugation and then cryopre- 
served in liquid nitrogen until use. These normal mammary epithelial cells 
have been grown in a complex medium which is a mixture of F12 and DME (1:1) 
supplemented with 5% fetal bovine serum, 4.5 g/L glucose, 5 /ig/ml insulin, 5 
/ig/ml epidermal growth factor, 5 fig/ml cholera toxin, 5 x 10~-^*^ M hydrocorti- 
sone, estradiol, dihydrotestosterone and triiodothyronine and then mixed with 
an equal volume of conditioned medium obtained from confluent cultures of human 
fetal epithelial cells from intestine or human adult bladder. One problem is 
that normal cells soon become senescent. Therefore, the supply of these cells 
is limited. Conditioned media from some hvmian tumor cell lines have been used 
since tumor cell lines do not show senescence and the supply is unlimited. 
Various human tumor cell lines were established from heterotransplanted human 
tumors in athymic mice which include five colon carcinoma lines, four lung car- 
cinoma cell lines, an astrocytoma and pheochromocytoma cell lines, and a mam- 
mary tumor cell line. All tumors were from different patients. These human 
tumor cells were grown to confluency and conditioned media were prepared to 
determine their effects on the growth of normal human mammary cells. The con- 
ditioned media from human colon carcinoma and human lung carcinoma cell lines 
seem to be satisfactory. These human carcinoma cell lines require a low con- 
centration of seriim (0.5%), when supplemented with hormones and factors; there- 
fore, attempts will be made to eliminate serum completely. Factors in the 
serum-free conditioned media will be characterized to determine which are best 
for supporting growth of normal human mammary epithelial cells in culture. 



Project Officer: Chester V. Piczak, B.S. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



177 



CONTRACT RESEARCH SUMMARY 

Title: Therapy of Patients with Stage II Carcinoma of the Breast 

Principal Investigator: Dr. Armando Guiliano 

Performing Organization: University of California 

City and State: Los Angeles, CA 

Contract Number: NOl-CB-43917 

Starting Date: 6/17/74 Expiration Date: 6/16/81 

Goal: To evaluate in patients with Stage II carcinoma of the breast the effec- 
tiveness of postoperative adjuvant chemotherapy and two forms of adjuvant 
chemo immunotherapy . 

Approach: Patients with positive axillary lymph nodes were randomly assigned 
to one of the following treatment schedules: (1) cyclophosphamide, methotrex- 
ate, and 5-fluorouracil (CMF); (2) CMF plus BCG; or (3) CMF plus BCG plus tumor 
cell vaccine (TCV). The tumor cell vaccine consisted of irradiated allogeneic 
breast carcinoma cells grown in tissue culture. Chemotherapy is administered 
in 12 cycles for 64 weeks. Immunotherapy was given concurrently and for one 
year following chemotherapy. BCG is administered by Tine technique; tumor cell 
vaccine was administered intradermally. Treatment failure consists of either 
metastases or development of carcinoma in the contralateral breast. The pa- 
tients are evaluated before treatment and at intervals throughout the study. 

Progress: One hundred thirty-one patients have been entered into the protocol 
since June 1974. The longest follow-up is 79 months. The recurrence rate for 
the CMF group is 6/37 (16.2%) at a 43-month mean follow-up. This group is 
being carefully studied to determine if prognostic factors are the same as in 
the CMF -BCG and CMF-BCG-TCV group. The recurrence rate for the CMF + BCG group 
is 23/53 (43%) and for the CMF + BCG + TCV group, 14/41 (34%) at a 59-month 
mean follow-up. These figures are all significantly better than the recurrence 
rate for untreated historical controls. There is no significant difference for 
recurrence and survival between the pre- and postmenopausal groups. 



Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: 



178 



i 



CONTRACT RESEARCH SUMMARY 

Title: Epidemiology of Benign Breast Disease 

Principal Investigator: Dr. Gary H. Spivey 

Performing Organization: University of California 

City and State: Los Angeles, CA 

Contract Number: NOl-CB-74202 

Starting Date: 8/15/77 Expiration Date: 8/14/82 

Goal: To examine the epidemiology of major types of benign breast disease and 
to compare identifiable risk factors for benign diseases with known risk factors 
for breast cancer. To examine a histologic classification scheme based on grad- 
ing of cellular atypia in comparison to the conventional histologic classifica- 
tion. 

Approach: Women with first biopsies for benign breast disease in participating 
hospitals are identified from pathology records and admitted to the study. They 
are selected to maximize representation of various histologic subtypes. Biopsy 
slides of participants are reviewed by two pathologists and classified according 
to two classification systems. A subcategory of women with breast cancer is 
included. Two types of controls are used, a hospital control and a friend con- 
trol. Information on epidemiologic characteristics is obtained by a question- 
naire covering a wide range of topics including environmental influences; 
familial cancer patterns; medical, maturation, and reproductive history; and 
other endocrine-related subjects. Epidemiologic characteristics of different 
histologic types of benign breast disease will be compared to each other and to 
those of breast cancer. 

Progress: Identification, recruitment, and interview of study subjects are 
almost complete. To date, 2,190 subjects have been interviewed, 121 have been 
assigned to interview, and 76 are still being recruited. We are currently 
coding, keypunching, and cleaning the remaining data and beginning preliminary 
analyses on a subset of the study population. At this time we have 1,879 ques- 
tionnaires on-line and cleaned. 



Project Officers: Elizabeth P. Anderson, Ph.D., B. J. Stone, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: $22,500 



179 



CONTRACT RESEARCH SUMMARY 

Title: Prediction of Hormone Dependency in Human Breast Cancer 

Principal Investigator: Dr. Elwood V. Jensen 

Performing Organization: University of Chicago 

City and State: Chicago, XL 

Contract Number: NOl-CB-43969 

Starting Date: 6/16/66 Expiration Date: 6/15/82 

Goal: To develop and improve techniques that will increase the predictability 
of response of human breast cancers to endocrine therapy. 

Approach: Since 1966, Dr. Jensen has been studying estrogen receptors (ER) in 
human breast cancer tissues and the relationship between ER content and response 
to endocrine manipulative treatments. Now that the predictive value of ER mea- 
surements has been established, attention is being directed toward development 
of simple, inexpensive assay procedures for ER in breast cancers and evaluation, 
of the ability of an ER assay on the primary tumor, carried out at the time of 
mastectomy, to predict subsequent response to endocrine therapy. 

Progress: Fusion of mouse myeloma cells with splenic l3nnphocytes from a Lewis 
rat, immunized with purified estrogen receptor from MCF-7 cancer cell cytosol, 
yielded hybridomas secreting antibodies to human estrophilin. After cloning by 
limiting dilution, three hybridoma cell lines were isolated, each secreting a 
monoclonal antiestrophilin antibody that recognizes a different antigenic deter- ^| 
minant on the receptor molecule. A combination of two of these antibody prepa- 
rations, one used to adsorb the receptor on polystyrene beads and the second 
labeled with 125]; ^q measure the amount of receptor thus bound, provides an » 
effective sandwich system for the radioimmunometric assay (IRMA) of estrophilin ■ 
in breast cancers. In a preliminary evaluation with ten human breast cancer ™ 
cytosols, the same relative receptor contents were found with the IRMA procedure 
as by sedimentation in sucrose gradients, although the absolute values obtained 
by the immunochemical technique were somewhat higher. The controlled pore glass 
bead procedure also has been compared with sucrose gradient analysis for more 
than 300 human breast cancer cytosols; again the same relative receptor contents 
were observed with the CPG method giving the same or, in some cases, higher ab- 
solute values. Clinical data abstraction for the breast cancer patients whose 
tumors were analyzed at the University of Chicago from 1966 to 1976 is in its 
final stages, and our systematic correlation of multiple parameters to provide 
maximum information will soon be feasible. Meanwhile, summaries of clinical 
abstracts are being furnished to the National Cancer Institute at the rate of 
about ten each month. 



Project Officers: Mary E. Sears, M.D., Ihor J. Masnyk, Ph.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: $25,000 



180 



CONTRACT RESEARCH SUMMARY 

Title: Detection of Immune Complexes in Sera of Patients with Breast Cancer 

Principal Investigator: Dr. M. Edward Medof 

Performing Organization: University of Chicago 

City and State: Chicago, IL 

Contract Number: NOl-CB-84224 

Starting Date: 9/1/78 Expiration Date: 8/30/81 

Goal: To assay sera from patients with breast cancer, benign breast disease, 
and normal volunteers for immune complexes (ICs) employing multiple techniques 
for IC quantitation, then to correlate levels where applicable with presence 
or absence of tumor (or benign disease), tumor progression, response to thera- 
peutic intervention and clinical course; to determine how hormonal status, past 
history and other clinical factors influence results, and to establish the 
relationship between IC data and pathologic features and other laboratory 
parameters. 

Approach: Four detection techniques for ICs (Raji cell assay, conglutinin- 
binding assay, solid phase Clq assay, and Clq-PEG precipitation assay) are 
established in this laboratory and will be run on sera from new patients with 
breast cancer (20-30) and benign breast disease (40-50). Serial samples from 
patients receiving and not receiving chemotherapy and/or Tamoxifen following 
mastectomy will be studied. Efforts will be made to obtain sera from other 
investigators in the breast cancer study group, assay these sera, and correlate 
IC data with other markers. Serum samples will be assayed for soluble and in- 
soluble, complement bearing and IgG-Fc accessible ICs. Data will be correlated 
with type of mastectomy, stage of patients, histologic type of tumor, including 
estrogen receptor positivity, histology of regional lymph nodes, as well as 
presence or absence of CEA and K-casein, fl-human chorionic gonadotropin (fl-HCG) 
and prolactin in selected cases. 

Progress: Systems are operational for collection, coding, and storage of blood 
samples; assembling pertinent clinical data; and for data analysis. IC assays 
have been perfected and standardized. Measurements have been made on sera from 
more than 400 patients. Epidemiological analyses of the various patient popu- 
lations have been made, including: analysis according to age, race, age at 
menarche and menopause, where applicable, pregnancy data and risk, factors. 
Risk factors include family history of cancers, duration of nursing, years of 
birth control pills, use of postmenopausal estrogens and exposure to smoking 
and carcinogens. Correlative analyses between IC levels and clinical data and 
other laboratory parameters have been made. Analyses so far indicate that 1) 
the Raji assay is most effective in differentiating cancer and noncancer 
groups, 2) ICs are present in certain types of benign disease, 3) there may be 
a correlation with estrogen receptor content, and 4) IC levels are affected by 
certain types of past history. 



Project Officers: D. Jane Taylor, Ph.D., Donald E. Henson, M.D, 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



181 



CONTRACT RESEARCH SUMMARY 

Title: Role of Dietary Factors and Non-contraceptive Estrogens in Breast 
Cancer 

Principal Investigator: Dr. Abraham M. Y. Nomura 

Performing Organization: Cancer Center of Hawaii 

University of Hawaii 
City and State: Honolulu, HI 

Contract Number: NOl-CB-53884 

Starting Date: 6/30/75 Expiration Date: 6/29/81 

Goal: To determine in postmenopausal women if there is an association of 
breast cancer with specific dietary factors and/or the use of menopausal 
estrogens. 

Approach: A case-control approach is being utilized with a personal interview 
to collect information on past history of drug usage (including menopausal 
estrogens) and usual weekly dietary intake. Breast cancer cases from three 
selected populations (ages 45-74) are identified: (1) the Caucasians in 
Hawaii, who are at high risk for breast cancer; (2) the Japanese in Hawaii, who 
are at intermediate risk; and (3) the Japanese in Fukuoka, Japan, representing 
a low-risk population. For each case, a neighborhood and hospital control, 
matched by race and age, is interviewed. Attempts are made to verify the drug 
history with the subject's personal physicians, and a subsample of the study 
population receives a repeated dietary interview to assess its reliability. 

Progress: The study originally aimed to interview 200 cases and 400 controls 
in each of the three ethnic groups. The Fukuoka portion of the study has been 
completed with the recruitment of 213 cases, and their respective hospital and 
neighborhood controls. The percentages of refusals were 1%, 4%, and 13% for 
cases, hospital controls, and neighborhood controls, respectively. In Hawaii, 
interviews have been completed on 187 cases, 180 hospital controls, and 177 
neighborhood controls among the Japanese and 163 cases, 159 hospital controls, 
and 141 neighborhood controls among the Caucasians, as of October 31, 1980. 
The current percentages of refusals are as follows: 15%, 18%, and 14% for the 
Japanese and 18%, 28%, and 15% for the Caucasian cases, hospital controls, and 
neighborhood controls, respectively. 



Project Officers: Elizabeth P. Anderson, Ph.D., David Levin, M.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: 



182 



ill 



CONTRACT RESEARCH SUMMARY 

Title: Effect of Chemotherapy-induced Endocrine Alterations on Stage II Breast 
Cancer 

Principal Investigator: Dr. Tapas K. DasGupta 

Performing Organization: University of Illinois 

City and State: Chicago, IL 

Contract Niamber: NOl-CB-84221 

Starting Date: 7/1/78 Expiration Date: 6/30/81 

Goal: To determine if the prolongation of the clinically disease-free interval 
produced by adjuvant chemotherapy is a result (1) of direct anti-cancer actions, 
(2) indirect action via endocrine organ changes, and (3) combination of 1 and 2. 

~ Approach: In the clinical study, premenopausal women with operable stage II 
breast cancer are registered to obtain preoperative hormone profiles. If 1 or 
more axillary nodes contain histologically proven metastases, the patient enters 
the adjuvant chemotherapy program. Estrogen (ER) and progesterone (PGR) recep- 
tor assays are performed. Study patients receive 12 courses of cyclophosphamide 
(Cytoxan), methotrexate, and 5-fluorouracil (5-FU). Blood samples are drawn 
during the first 2 weeks of each 28 day cycle of chemotherapy for 12 consecutive 
cycles during CMF administration and every 3 months thereafter to determine hor- 
mone levels. In animal studies 1-methyl-l-nitrosourea (NMU) is used to induce 
mammary tumors in BUF/N rats. The effects of CMF on tumor dynamics and hormone 
levels will be determined. 

Progress: To date 3 of 34 (9%) evaluable patients receiving CMF chemotherapy 
have failed. None have completed treatment. All were ER~. Table 1 shows the 
incidence of ER and PGR in stage II patients. Of 34 tumor cytosols 26 (76%) 
have been assayed for PGR. Nineteen of 34 tumor cytosols (56%) were ER"^ (>3 
fm/mg protein) . 

Table 1. Receptor Distribution in Stage II Breast Cancer Cytosols 

+ + +- -+ -- +NA - NA 

ER PGR ER PGR ER PGR ER PGR ER PGR ER PGR 



7/26 (27%) 8/26 (31%) 0/26 11/26 (42%) 4/8 (50%) 4/8 (50%) 

NA: Not assayed 

Tumor and cytosol ER incidence and txmior latency in BUF/N rats are depen- 
dent on the day of the estrous cycle NMU is administered. Ovariectomy produced 
a complete response (25%) and partial response (47%) within 2 weeks. As a sin- 
gle agent, only Cytoxan produces a significant alteration in estrous cyclicity 
and reduction in tumor growth taken as a percentage of pre treatment volumes. 
Cytoxan does not significantly affect tumor growth in ovariectomized rats. The 
effect of Cytoxan, methotrexate and 5-FU as single agents and in combination 
on tumor growth and hormone levels in intact and ovariectomized animals is cur- 
rently being evaluated. 

Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: 

183 



Title; 



CONTRACT RESEARCH SUMMARY 

Benign and Non-invasive Breast Lesions in Groups at Different Risk for " 
Breast Cancer 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Sue Amelia Bartow 
University of New Mexico 
Albuquerque, NM 



Contract Number; 
Starting Date: 



NOl-CB-84231 
9/30/78 



Expiration Date: 9/29/82 



Goal: To investigate whether population groups with low incidence of breast 
cancer also have a low prevalence at autopsy of benign breast lesions. 

Approach: Combining the resources of the population-based New Mexico Tumor 
Registry, the State's Medical Investigator's Office and the Medical School 
Departments of Pathology and Radiology, we propose to examine and compare benign 
breast lesions in three ethnic groups in an autopsy series — Anglos, Spanish 
Americans, and American Indians. These three groups are all at differing risk 
for breast cancer. The incidence rates and histopathologic distributions of 
breast cancers are already known from Tumor Registry data. The autopsy series 
to be examined will include about 500 women in three years, 230 Anglo, 140 
Spanish American, and 130 American Indian. Breast lesions will be classified 
pathologically. Radiological studies of the specimens by both clinical mammog- 
raphy and high resolution techniques will be compared with histopathological 
findings. Medical, reproductive, and related history of each case will be ob- ^ 
tained from the family physician or medical records if at all possible. ■ 

Progress: A total of 212 cases have been accessioned into the study (127 Anglo, 
52 Spanish, and 33 American Indian). Autopsies are now being preferentially 
performed on the low risk Spanish and Indian women. Medical histories on 182 
cases include 80 complete interviews, 5 incomplete, 39 pending, and 58 unable 
to complete. Radiologic analysis of 178 cases indicates the following: preval- 
ence of "high risk" patterns by Wolfe's classification is greater , in the Anglo 
than in the Indian population; high risk patterns are more often present in the 
>40 age group in Anglo women than in Spanish/ Indian women. Morphologic analysis 
of 100 cases showed the following trends: earlier and more complete involution 
of the non-fat breast tissue in Indian and Spanish women as compared to Anglo 
women; presence of cystic change and ductal epithelial proliferative changes in 
all three ethnic groups, increasing with age. Correlation of morphologic find- 
ings with breast parenchymal patterns of Wolfe's mammographic classification 
groups shows the patterns to be the result of varying degrees of periductal and 
intralobular fibrosis. Presence of ductal epithelial proliferative changes does 
not appear to correlate significantly with the "higher risk" mammographic breast 
patterns. Computerization of the data, now under way, will be necessary to ana- 
lyze more completely the data being collected. 



Project Officers: Elizabeth P. Anderson, Ph.D., Louise Brinton, Ph.D. 
Program: Breast Cancer Epidemiology 
FY 81 Funds: $30,000 



184 



f 



CONTRACT RESEARCH SUMMARY 

Title: Methods to Predict Chemotherapy Sensitivity 

Principal Investigator: Dr. Russell Hilf 

Performing Organization: University of Rochester 

City and State: Rochester, NY 

Contract Number: NOl-CB-74204 

Starting Date: 7/15/77 Expiration Date: 7/14/82 

Goal: To develop a reliable prognostic test that will predict the efficacy of 
chemotherapeutic agents in the treatment of individual breast cancer patients. 

Approach: This project will perform a battery of selected enzymes in primary 
and metastatic breast cancer specimens and will develop mathematical models 
based on the biochemical profile. The enzymes include lactate dehydrogenase, 
pyruvate kinase, glucosephosphate isomerase, isocitrate dehydrogenase, phospho- 
glucomutase and glucose 6-phosphate dehydrogenase; DNA and protein levels will 
be measured. Hormone receptor assays and histopathology will also be performed. 
Data concerning the patients' biological and oncological histories will be 
acquired, evaluated and stored. Analyses of the combined data will test the 
predictive accuracy of the mathematical model constructed from the biochemical 
profile. 

Progress: Accession of cases for the project continues at the anticipated rate 
as follow-up data on response or recurrence are obtained and entered into the 
computer file from the forms devised for this study. More than 350 cases have 
been identified as eligible according to the criteria established at the outset. 
A logistic regression model has been developed, based on the activities of LDH 
and the product of the activities of PK x GPI x ICD. We have 93 patients with 
advanced disease, classified according to a 50% probability estimate as the cut- 
off for response vs. no response. For non-re sponders, the model correctly clas- 
sified 58/61 (95%) and for responders 22/32 (69%), an overall accuracy of 80/93 
(86%). We have identified 82 patients receiving adjuvant therapy; 54 have had 
recurrences. Patients recurring have demonstrated low enzyme activity profiles. 
We have now applied a statistical method of Cox in which biochemical parameters 
act as regressor variables in the hazard function and are related to time to 
recurrence. Employing maximum log-likelihood criteria for model selection, we 
have developed models that gave a negative coefficient for GPI indicating that 
higher GPI activity suggests decreasing hazard, i.e., recurrence. Additional 
factors were found to be menopausal status. A further extension to patients 
receiving no therapy (about 160 cases) is under way and here we are finding that 
enzymes and DNA may have prognostic implications. In each of these clinical 
groups, ER status does not appear to offer prognostic value for disease outcome 
in patients receiving cytotoxic chemotherapy. 



Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: 

185 



CONTRACT RESEARCH SUMMARY 
Title: Biochemical Mechanism of Endocrine-induced Breast Cancer Regression ^ 



Principal Investigator: 
Performing Organization: 



City and State: 

Contract Number; 
Starting Date: 



NOl-CB-23862 
6/1/72 



Dr. William L. McGuire 

University of Texas Health Science 

Center 

San Antonio, TX 



Expiration Date: 5/31/82 



Goal: To obtain new information of the hormonal control of breast tumor growth 
and to identify those breast cancer patients whose disease will respond to hor- 
monal manipulation. 

Approach: To provide correlations of estrogen and progesterone receptor mea- 
surements with prognosis at the time of mastectomy and response to endocrine 
therapies. To evaluate histochemical methods of measuring steroid receptors. 

Progress: Clinical correlations of steroid receptor in primary breast cancer 
and prognosis. — We have completed our double blind study with Dr. Ed Fisher 
in Pittsburgh correlating ER content and histopathologic variables. We find 
several histologic variables that correlate with estrogen receptor status. 
Highly statistically significant correlations between ER content and histologic 
grade and nuclear grade, tumor necrosis, elastosis and Ijmiphoid cell infiltra- 
tion were observed. We conclude that ER is a biochemical marker for the degree 
of differentiation of hiiman breast cancer providing in part a rationale for the 
observed differences in biologic behavior between receptor positive and nega- 
tive tumors. 

We are concentrating our efforts in obtaining clinical follow-up data in pa- 
tients in whom we have assayed ER and PgR. The coming year will be devoted to 
increasing the follow-up and analyzing the data. 

Clinical Follow-up of Breast Cancer Data 



San New 

Antonio Downstate Cleveland Orleans 



All cases assayed for receptor 
# cases with clinical follow-up 



1718 
1095 



676 
201 



1357 
1037 



311 
178 



Total 

4062 

2511 

62% 



Immunocytochemistry — We have tested probes that might detect ER using cyto- 
chemical techniques. So far none is successful and we have found many that 
other people use are useless. Recently we have obtained some new steroid con- 
jugates that might avoid some of the current difficulties and are currently 
testing them. 



Project Officer: Mary E. Sears, M.D. 
Program: Breast Cancer Treatment 
FY 81 Funds: -$112,600 



186 



CONTRACT RESEARCH SUMMARY 

Title: Risk Associated with In Situ Carcinoma and "Precancerous" Maranary 
Hyperplasias 



Principal Investigator: 
Performing Organization: 



City and State: 

Contract Number: 
Starting Date: 



NOl-CB-74098 
7/15/77 



Dr. David L. Page 

Vanderbilt University School of 

Medicine 
Nashville, TN 



Expiration Date: 4/14/81 



Goal: To compare subgroups of females regarding their incidence of breast car- 
cinoma during a 10 to 25 year interval after biopsy diagnosis of ^n situ carci- 
noma and mammary epithelial hyperplasias. 

Approach: This will be a retrospective cohort study in which slides have been 
reviewed and classified on over 11,000 breast biopsies performed during 1952- 
1968. The study population with proliferative disease will then be contacted 
and follow-up information on each subject obtained regarding subsequent devel- 
opment of mammary carcinoma, as well as other pertinent clinical and epidemio- 
logical data. The control group will be composed of patients with biopsy but 
no evidence of proliferative lesions. An effort will be made to identify those 
specific breast lesions or combinations of lesions which are associated with 
an increased risk of development of breast cancer. Non-anatomic risk factors 
will also be correlated. 

Progress: There have been 3,519 contact letters mailed with 2,807 completed 
questionnaires on file. Operating surgeons and their office personnel have 
given invaluable aid in these efforts. A data quality assurance program has 
been designed and implemented. Two programs for statistical analysis have been 
written and are in place for final analysis. Histologic classification is com- 
plete for all biopsies in the study. The total number of biopsies with hyper- 
plastic lesions or carcinoma in situ is 2,735. One hundred sixty-six biopsies 
are identified with carcinoma in situ . One hundred sixty biopsies with atypi- 
cal lobular hyperplasia and 690 biopsies with severe hyperplasia of ductal type 
are identified. There are 1,418 biopsies demonstrating ductal hyperplasia of 
moderate degree. 

We have completed review and follow-up of 28 patients with ductal carcinoma 
in situ treated by diagnostic biopsy only. Three of these patients were fol- 
lowed less than three years, dying of causes unrelated to breast disease or 
treated by bilateral mastectomy. Of the remaining 25 patients treated with 
biopsy only, 28% developed invasive breast carcinoma with an average follow-up 
of 16 years. All invasive breast carcinomas appeared in the same breast demon- 
strating in situ carcinoma in the biopsy. Six of these were in the same qua- 
drant and one was in an adjacent quadrant. The average interval to invasive 
carcinoma development was 6.1 years (range 3 to 11 years). The average age at 
biopsy for all women was 52. Four of the seven women developing invasive car- 
cinoma have developed distant metastases, with three of them dying of metastatic 
breast cancer. 

Project Officer: D. Jane Taylor, Ph.D. 
Program: Breast Cancer Diagnosis 
FY 81 Funds: 



187 



CONTRACT RESEARCH SUMMARY 

Title: Biologic Markers in Breast Cancer: Patient Resource 

Principal Investigator: Dr. Leslie W. Whitney 

Performing Organization: Wilmington Medical Center 

City and State: Wilmington, DE 

Contract Number: NOl-CB-74137 

Starting Date: 9/1/77 Expiration Date: 8/31/81 

Goal: To develop a Breast Cancer Task Force specimen resource for blood from 
breast cancer patients and controls to be used in a search for and verification 
of new breast cancer markers. 

Approach: Thirty milliliters of blood were collected from all project partici- 
pants with malignant breast lesions and all benign patients who developed malig- 
nant breast lesions during the first year follow-up. These specimens are pro- 
cessed, shipped, and stored at -70 °C at an NCI-designated blood bank facility 
with appropriate clinical data. 

Progress: In the first contract year (9/1/77 - 8/31/78) 65 patients were en- 
tered into the study, 27 of whom have malignant disease. From these 27 malig- 
nant patients we have completed a 3-year follow-up on 21. Of the remaining 6, 
2 have expired, 1 has moved out of state, 1 refused, and 2 are unable to have 
blood drawn at the present time. In the second contract year (9/1/78 - 8/31/79), 
248 patients were entered, 80 of whom have malignant disease. We have obtained 
a 2-year follow-up on 67 malignant patients; of the remaining 13, 5 have refused ■ 
6 have expired and we are unable to contact the remaining 2. 

For the 206 benign patients in the studdy, a follow-up questionnaire with 
cover letter was sent to all. A 1-year follow-up was completed on 198 patients 
either by direct mail return, telephone contact with the patient if no mail 
return, or contact with doctor's office or hospital clinic if we were unable to 
contact the patient. One patient refused follow-up and 7 had no doctor contact 
and have moved leaving no foirwarding address. 

From 575 patients, 5,917 vials of serum have been shipped to the storage 
facility at Mayo Clinic. 






Project Officers: Ihor J. Masnyk, Ph.D., Mary E. Sears, M.D. 

Program: Breast Cancer Treatment im'\ 

FY 81 Funds: 



I 



CONTRACT RESEARCH SUMMARY 

Title: Produce and Identify Antibodies to Collagens/Procollagens and/or 
Related Enzymes 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Heinz Furthmayr 
Yale University 
New Haven, CT 



Contract Number: 
Starting Date: 



NOl-CB-84225 
9/18/78 



Expiration Date: 9/17/81 



Goal: To prepare immunochemically defined antibody reagents to collagen of 
types I, II, III, IV (basement membrane), AB (fetal collagen), procollagens of 
types I and III and mouse collagen of types I, III and IV. 

Approach: Studies include: (1) Isolation and purification of collagens from 
calf, human, and mouse tissue for immunization, affinity columns, and serolog- 
ical testing; 2) immunization of rabbits and mice; 3) serological screening of 
antisera during immunization and after final bleeding; 4) obtaining monoclonal 
antibodies to type IV collagen and type I and III procollagen by the somatic 
cell hybridization technique; (5) isolation and purification of antibodies spe- 
cific for each collagen; and (6) characterization of the isolated antibodies. 

Progress: In addition to progress reported in the previous two years, small 
amounts of the aminoterminal propeptide of type I and type III collagen were 
obtained to be used for producing monoclonal antibodies. Type IV collagen 
(pepsin extract) from calf lung and human placenta and a fragment (E-chain) 
presvimably derived from basement membrane collagen were isolated. The prepara- 
tions, in addition to biochemical characterization, were analyzed by the new 
technique of rotary shadowing, which has been introduced in the laboratory 
recently. Groups of 10 rabbits each were immunized with mouse collagen type I 
and type III and pure antibodies were isolated to mouse type I thus far. Cur- 
rently, work is progressing on the isolation of mouse type III antibodies. 
Antibodies were produced and isolated to the "E-chain," which react with base- 
ment membranes in immunofluorescence experiments. During the past eight months 
they have been working on the application of the monoclonal antibody technique 
and have obtained 9 clones of cells which synthesize IgM antibodies to calf 
type IV (lung) collagen and four of these monoclonal antibodies react by tissue 
fluorescence. These antibodies at present do not appear to cross-react with 
human basement membranes as shown by serological or by immunofluorescence 
tests. Characterization of monoclonal antibodies to type IV collagen from 
human lung is being done. Hybrid cells were produced which synthesize anti- 
bodies to human type V (AB2) collagen. These hybrid cells have not yet been 
cloned, however. Mice were immunized with propeptide type I and the first 
fusion experiment is planned in the near future. 



Project Officer: Chester V. Piczak, B.S. 
Program: Breast Cancer Experimental Biology 
FY 81 Funds: 



189 



190 



CANCER DIAGNOSIS RESEARCH PROGRAM 



DESCRIPTION 

The Cancer Diagnosis Research Program emphasizes research in early detec- 
tion, diagnosis (which includes staging and prognosis), tumor localization, 
and monitoring the changes during therapy or progression of disease. The pro- 
gram also seeks to apply the knowledge obtained to appropriate populations for 
clinical evaluaation. Projects in these areas are frequently concerned with 
improvement of existing techniques as well as the development of new tests and 
procedures. Many of the projects in the Cancer Diagnosis Research Program have 
begun in a more basic area such as, Tumor Biology, Tumor Immunology, or General 
Medical Sciences for instrument development. As a basic concept becomes poten- 
tially useful in one of the areas of cancer diagnosis the project may be trans- 
ferred to the Cancer Diagnosis Research Program. In the earliest stages, diag- 
nostic research is not necessarily site (or even disease) oriented. 

The major objective of the Program is to recognize or detect cancer at the 
earliest possible stage to allow appropriate therapy to begin. Early detec- 
tion and early treatment should improve the chances for the control of cancer, 
decrease mortality from the disease and increase survival and quality of life 
of those with cancer. Additionally, early detection is providing greater un- 
derstanding of the natural history of different types of cancer in the early 
stages of disease. 

Since the techniques of cancer detection and diagnosis are useful for many, 
or all, types of cancer the divisions of the program are not by cancer type or 
organ site, but by technical discipline. The Diagnosis Research Program con- 
sists of projects in eight disciplinary categories: Biochemistry, Immunodiag- 
nosis, Cytology, Pathology, Radiological Imaging, Non-Radiological Imaging 
(Ultasound, Nuclear Magnetic Resonance, Thermography and Microwave), Nuclear 
Medicine and those projects that are clearly Multiple Disciplinary. 

The distribution of contracts and grants currently included (FY1980) in 
the Diagnosis Research Program into the various categories in summarized in 
the following table. Each category is discussed in the sections following the 
table. 



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192 



CANCER DIAGNOSIS RESEARCH PROGRAM 
1. BIOCHEMISTRY 

Biochemical methods to improve diagnosis and detection of cancer involve the 
study of a variety of substances such as hormones, enzymes, proteins and meta- 
bolic products in the circulation and in other biological fluids, as well as 
the study of surface characteristics of tumor cells and chemical characteri- 
zation of tumor cells. 

Under hormonal studies, the measurement and mode of action of the multiple 
forms of human growth hormone are being studied under grant CA-14025. The 
objective of grant CA-23848 is to develop a simple and rapid method of detec- 
ting estrogen receptors in the endometrium and mammary gland using fluorescein- 
labeled hormones and fluorescence microscopy. The results are being correlated 
with the clinical response of the patients to endocrine therapy. Identifica- 
tion, isolation and characterization of thyroid hyperfunction in patients with 
trophoblastic tumors, especially hydatidiform mole, are being pursued under 
grant CA-31218. A series of experiments have been proposed in grant CA-29062 
to evaluate the clinical use of a material which is cross-reactive to vasoactive 
intestinal peptide (VIP) as a biochemical marker to refine the diagnosis of 
acute leukemias. 

A variety of enzymes are being studied for their correlation with different 
cancers. Histaminase and an isozyme of alkaline phosphatase (FHAP) are being 
investigated as general cancer markers in grant CA-26652 and contract CB-74173, 
respectively. The biochemistry of isozyme 5 of acid phosphatase and its clini- 
cal implications in the diagnosis and prognosis of prostatic cancer and of 
hairy-cell leukemia will become topics of investigation for a new grant CA- 
31187. Studies of 5'-nucleotide phosphodiesterase which has been shown to be 
elevated in patients with known hepatic metastases from mammary carcinoma will 
be continued under grant CA-25376. A basis for characterizing and classifying 
human lung tumors and revealing essential biochemical pathways vulnerable to 
chemotherapy by means of quantitative enzyme profiles is being sought under 
grant CA-25016. Enz3rmes are being determined in plasma, blood cells and liver 
biopsy samples in grant CA-22065 to ascertain their predictive value in diag- 
nosis and in monitoring of lymphoma patients during therapy. Serum, urine and 
CSF ribonucleases in tissues and body fluids of healthy subjects and breast 
cancer patients will be isolated, characterized and correlated with disease 
status under grant CA-19606. In grant CA-25548 human tumor cell lines will be 
characterized by isozyme phenotyping to confirm whether they are bona fide 
representatives of the tumors from which they were derived. Differentiation 
of lymphocytes by the enzyme marker, terminal deoxynucleotidyl transferase, 
(TdT) is being investigated under grant CA-22599 by determination of the re- 
lationship of presence or absence of the enzyme to particular cell sets and to 
tumors representing those cell sets. 

Proteins as discriminants of cancer are being investigated under several 
grants. Protein analysis of pancreatic secretions is being probed for diag- 
nostic potential in pancreatitis and pancreatic cancer under grant CA-14380. 
Most of the research effort in grant CA-19634 is devoted to a characterization 
of lipoprotein differences, qualitative and quantitative, between mammary car- 
cinoma patients, normal individuals with a family history of cancer and those 



193 



with a negative family history of breast cancer. Intercellular matrix proteins 
are being extracted from human chondrosarcomas in grant CA-23945 and compared 
with normal articular cartilage to evaluate degree of malignancy and possibly 
grade chondrosarcomas biochemically as well as morphologically. A new investi- 
gator research grant CA-30667, recently awarded, aims to exploit the differ- 
ences in lectin binding characteristic of the mucin found in normal colon and 
abnormal colon, malignant and premalignant , to predict the risk of developing 
colon cancer. 

Urinary nucleoside breakdown products of tRNA are being measured in cancer 
patients in grant CA-25210 to explore the molecular mechanism responsible for 
increased excretion in cancer patients. In grant CA-14185 modified nucleosides 
derived from the turnover of human RNA and from the nucleic acid anabolic pro- 
cesses and present in urines of cancer patients will be evaluated as possible 
quantitative tumor markers. The values of guanosine monophosphate (cyclic 
GMP), and adenosine monophosphate (cyclic AMP) in plasma and urine of patients 
with precursor lesions and gynecologic neoplasms are being determined in grant 
CA-25357 to ascertain their usefulness in screening, detection, diagnosis, 
treatment and follow-up of patients with gynecologic neoplasms. 

A "myasthenic" substance found in small cell lung cancer tissue is being 
studied as a possible aid in detection of lung cancer under grant CA-22885. 
The aim of CA-21958 is to compare quantitatively and qualitatively the serum 
heteroglycan fractions of tumor bearing animals with those from controls to 
ascertain whether these can be utilized for detection of malignancies and for 
monitoring of tumor burden. Electron paramagnetic resonance (EPR) measurements 
of cupric ion bound to ceruloplasmin are being made to ascertain whether these 
can indicate the presence of cancer in humans under grant CA-14335 and to cor- 
relate changes with success or failure of treatment. Gas chromatography of the 
colonic microbial metabolites in breath will be performed in grant CA-2905& to 
test the hypothesis that these metabolities may be useful markers for increased 
risk of developing colonic cancer. 

2. IMMUNODIAGNOSIS 

The portion of the Diagnosis Program classified as Immunodiagnosis can be 
subdivided into projects dealing with circulating tumor antigens or markers, 
such as oncofetal antigens, hormones, enzymes, and glycoproteins; projects 
dealing with tumor associated antigens, research in localization of tumors by 
radioimmunodetection; studies of lymphocytes in host-tumor relationships, and 
projects dealing with antibodies to tumors (including immune complexes). 

The carcinoembryonic antigen is the most widely utilized circulating onco- 
fetal tumor marker and contract CB-33848 deals with the prognostic and monitor- 
ing value of CEA following therapy of colorectal and lung cancer. Additionally 
there is a study on the heterogeneity of CEA, grant CA-24376, and comparison 
of new gastrointestinal cancer antigens with CEA and improvement of their 
clinical use in program project grant CA-04486. 

Several projects are investigating the relationship between hormone levels 
and cancer: calcitonin and 6-lipotropin are subjects for two grants, CA-22137 
and CA-23382, respectively. Two other studies of serum calcitonin as a screen 
for family members of patients with medullary thyroid cancer are supported by 



194 



grant CA-22595 and contract CB-639^94. The role of serum thyroglobulin levels 
as markers of tumor recurrence in thyroid cancer is being investigated under 
grant CA-25338. 

Two studies of enzymes as markers for cancer deal with measurement of serum 
levels of tyrosinase in patients with melanoma, grant CA-25381 and of UDP-galac- 
tosyltransf erase in patients with ovarian and breast carcinoma in contract CB- 
84260. The prostate is the organ site involved in contract CB-74169 evaluating 
acid phosphatase in a screening population and in grant CA-23990 testing acid 
phosphatase and ribonuclease as markers for the early detection of prostatic 
carcinoma. The objective of a new grant, CA-29225, is to develop an immuno- 
histochemical technique for the visualization of a monocyte esterase marker to 
be employed in the differential diagnosis of subtypes of leukemias. 

Proteins or their degradation products have been detected in body fluids 
and have demonstrated correlation with the presence of cancer. The role of 
SAA, a major protein constituent of secondary amyloid is being investigated as 
a marker of tumor recurrence, response to therapy and immune function in cancers 
of the lung and gastrointestinal tract on grant CA-22141. Gp 48, a major group 
of glycoproteins sythesized and released into organ culture from breast adeno- 
carcinoma specimen are being assessed for their diagnostic significance in 
grant CA-24645. A glycoprotein, EDCl , is being monitored in both urine and 
plasma of breast cancer patients and controls for its iramunodiagnostic value 
under contract CB-84308. Studies are being continued under grant CA-31762 to 
biochemically characterize a glycoprotein surface antigen found on myeloblasts 
of leukemia patients, prepare antiserum to it and monitor its efficacy in pre- 
dicting relapse of leukemia patients in remission. 

Tumor associated antigens studies are under way for many organ sites: 
breast, CA-20286; cervix and head and neck, CA-84267; urogenital tract, CA- 
.27213; melanoma, CA-30019; mesothelioma, CA-27081; colon, CA-26246, CB-84259, 
and CB-84257. Studies of nuclear antigens associated with normal and leukemic 
human blood cells to determine their usefulness as markers in hematopoiesis 
are being supported by grant CA-26948. 

Several studies are concerned with techniques for labeling antibodies with 
radionuclides for tumor detection and localization. Grant CA-30255 is directed 
towards localization of germ-cell tumors and hepatomas which produce alphafeto- 
protein by labeling of monoclonal antibodies with ^-^^lodine, ^'Technetium and 
^■'■llndium. An approach is being tested under grant CA-28462 to develop a better 
technique for labeling antibodies to human serum albumin (HSA) with ^^^In or 
^^Tc or ■'■^^I, and studying the effect of various conditions, of conjugation on 
accelerated blood clearance and retention of immunological activity. The 
objective of grant CA-17742 was to study conditions favoring the localization of 
CEA-containing human tumors in animal systems by means of radiolabeled anti-CEA 
immunoglobulins and total-body photoscanning. Grant CA-25584 is a continuation 
of the program of CEA-tumor radioimmunodetection and involves planning a clinical 
trial of patients with proven malignancy to further evaluate its use in initial 
tumor diagnosis and in the management and clinical staging of cancer patients. 
A new grant CA-29639 will concern itself with the development of a method for 
melanoma localization using radiolabeled monoclonal antibody fragments against 
p97 antigen and imaging by emission tomography. 



195 



Lymphocytes and monocytes in host tumor responses are the subject of 
several projects. The development of clinically useful diagnostic tests 
based on the selective binding of bacteria and antibody-coated bacteria to 
lymphocyte subpopulations is being undertaken under grant CA-29552. A 
contract, CB-84261, has been involved in an extensive effort to evaluate 
lymphoid differentiation antigens, particularly those on bone marrow and 
thymus cells, as potential immunodiagnostic markers for leukemia and lymph- 
omas. The relationship between the clinical status of cancer patients and 
the functional activity of their monocytes _in vitro is being determined under 
contract CB-74121. Contract CB-74131 is concerned with defining optimal cryo- 
preservation techniques of human monocytes to preserve antibody-dependent 
cellular cytotoxicity for immunologic studies. 

Circulating antibodies to tumor antigens are being studied in patients 
with melanoma, in contract CB-74120; while the presence of circulating immune 
complexes is being quant itated and analyzed in a number of cancers including 
colon, breast and melanoma under contract CB-84262. Antibodies are also 
serially measured in mice while developing induced tumors in contract CB-74134. 
Investigations of the expression of the major virion glycoprotein gp52 of MMTV 
on late-occurring mammary tumors in mice and its potential as a tumor marker 
will be pursued under a new grant, CA-28305. 

In addition to the research grants and contracts, there are contracts for 
the collection and storage of serum samples from patients with cancer and 
other acute and chronic diseases and during the course of therapy for certain 
cancers. These contracts, CB-33914, CB-04350, CB-74210, and CB-84258, have 
been invaluable for the rapid evaluation of a wide variety of serologic tests 
for cancer that come to the attention of NCI. CEA levels on blood stored at 
the NCI-Mayo Serum Bank are performed under contract CB-23854. A tissue cul- 
ture bank for cell lines which can be utilized by an investigator for research 
in cancer immunodiagnosis is maintained under contract CB-43854. 

3. CYTOLOGY 

Diagnostic cytology research projects include the development of automated 
instrumentation and cell markers that can be used to differentiate normal and 
atypical cells. Instruments undergoing development and testing are those of 
high resolution slide based, grants, CA-13271, CA-28833, CA-27313, and CA-31049 
and contracct CB-33873, and flow fluorometric types, contracts CB-40300 and 
CB-33862. Development, construction and testing of an ultrafast optcal scanner 
for microscopic specimens, grant CA-24466, will make possible the automated 
search for abnormal, transformed or cytochemically marked cells in microscopic 
preparations. Single cell classification algorithms employing the current 
state of the art in digital image processing, scene segmentation and image 
acquisition hardware are being developed and tested in contract CB-70314. The 
classification algorithm chosen with human interaction was found to perform at 
an acceptable error rate. A cost/utility analysis indicates that the chosen 
algorithms can operate with error rates adequate for routine screening. 

A flow instrument currently undergoing testing uses dual staining and 
analysis with a dual laser beam sorter, contract CB-40300. In addition, 
a slit-scan using a combined static and flow system is now being tested to 



196 



determine the system characteristics including rate and causes of false alarms, 
contract CB-33862 and grant CA-30582. Quantitative descriptors of normal and 
abnormal cells include cytochemical and immunological markers which increase the 
potential sensitivity and specificity of sensor systems. 

Two different systems are under development to place individual sorted cells 
on slides for subsequent retrieval and analysis, grants CA-28886 and CA-28706. 

Chemical synthesis of cytochemical probes, grants CA-28770 and CA-30148, 
with sharp fluorescence emission spectra will enhance multiple staining of cells 
for flow systems and for cells on slides in static systems. Cellular fluores- 
cent markers under investigation also include studies of fluorescent substrates 
for demonstration of cAMP phosphodiesterase, adenyl cyclase, and acid phospha- 
tase, grant CA-19552; acridine orange, contract CB-33862; and chromomycin A3, 
contract CB-40300. Cellular DNA content and size (as measured by orthogonal 
light scatter) have been shown to detect abnormal cells, contract CB-40300. 
Cell surface antigens may also be used as markers for abnormal cells. With 
appropriate fluorescent tagging of these antigens, normal and abnormal cells may 
be differentiated by automated instruments. Potentially useful markers being 
studied are Herpes simplex virus related antigens, grant CA-28724 and contract 
CB-74170, and nucleolar antigen, grant CA-28771. In addition, cytochemical and 
biophysical probes of nuclear and cytoplasmic structure are being used to dis- 
tinguish normal and malignant cells, grant CA-28704. 

Analysis of the performance of automated systems, comparison of instrumental 
or system classification and manual microscopic classification utilizing standard 
morphologic criteria is studied under contract CB-74190. In the same contrct, 
programming of cell recognition algorithms continues with emphasis on scene seg- 
mentation techniques. 

Flow cytometry is used in grant CA-27283 to develop an assay for detecting 
transformed cells (by chemical carcinogens) differentially labeled with fluor- 
escamine. In addition, the nature and functional significance of a hyaluroni- 
dase sensitive "barrier" surrounding transformed cells is being investigated. 

To better understand and treat lymphoproliferative disease, flow micro- 
fluorometry analysis of human lymphoid malignancies is under investigation, 
grant CA-23393, with the hope that malignant subpopulations can be identified 
by a two parameter analysis for size and surface immunoglobulin. In addition, 
it is hoped that the ploidy and cell cycle kinetic parameter of these cells 
will aid in the diagnosis, classification, scheduling and monitoring of treat- 
ment. A combined flow cytometric cell-sorting autoradiographic technique is 
being developed in grant CA-25348 to monitor acute leukemias of adults and 
children and to predict relapse and evaluate treatment protocol. Using fluor- 
escence microscopy and flow microf luorimetry in grant CA-27123, merocyanine 540 
is employed to study leukemia to characterize the dynamic changes in the stain- 
ing pattern during the clinical course and to isolate and characterize hemato- 
poietic progenitor cells. The selective staining of leukemic cells might be a 
useful adjunct to existing methods of monitoring the course of leukemia and of 
prognosis of relapse or remission. Merocyanine 540 is also utilized to eluci- 
date the relationship between dye binding and the transformed state of cells 
in grant CA-28921. Leukemic cells are also being studied to determine their 
and to test chemotherapeutic agents for their ability to alter patterns of 



19 7 



cell differentiation _in vivo , grant CA-22942. Study of human leukemic and 
preleukemic blood and bone marrow using the soft agar culture techniques has ( 
continued, grant CA-17353, to characterize the status and prognosis of hemato- 
prolif erative disorders. Measurements of the quality and quantity of colony 
formation appear to provide useful indicators of disease status and progress- 
ion as well as diagnosis and predictions concerning response to therapy of 
acute nonlymphocytic leukeraias and preleukemic states. 

In an effort to predict the biological potential of gynecologic neoplastic 
lesions, a quantitative analysis of nuclear DNA content using Feulgen micro- 
spectrophotometry is under way, grant CA-24932. Determination of the nuclear 
DNA changes may reveal the correlation of the changes with regression, persis- 
tence and progression of squamous cancer of the uterine cervix. Earlier find- 
ings of carcinoma-in-situ of lung, intestine, ovary and bladder might be possi- 
ble by using an immunocytologic technique being evaluated in grant CA-26863, 
to detect carcinoembryonic antigen on exfoliated cells. An extensive study to 
test the feasibility of screening for endometrial cancers in asymptomatic women 
by means of uterine sampling is being done under contract CB-84233. 

The Eighth Conference on Analytical Cytology was partially supported by 
grant CA-29859. 

4. PATHOLOGY 

A system of classifying human pituitary adenomas based on electron micros- 
copy and immunocytochemistry has been developed on grant CA-21905. To expand 
understanding of tumor behavior- in the pyriform sinus and the oral cavity, 
whole organ sections of patients who undergo surgical excision have been 
studied, grant CA-22101, using microscopic appearance to determine size and 
extent of tumor, relation of tumor spread to the laryngeal framework and 
specific routes of tumor spread. Malignant lymphomas and leukemias are being 
studied on grant CA-26422 by a combination of methodologies along with routine 
morphology to determine the most valuable tests to provide earlier detection and 
more precise classification. Disaggregation and dispersion of solid tumors so 
that specific cells of interest may be isolated and concentrated for character- 
ization is the goal of grant CA-23922. Contract CB-84257 utilizes immunohisto- 
chemical techniques to identify a variety of tumor associated antigens in tumor 
cells as a means for detection of metastatic disease in lymph nodes. 

5. RADIOLOGICAL IMAGING 

Several contracts and grants to improve x-ray imaging currently are under 
study. One contract, CB-74211, seeks to develop large area solid state image 
receptor for x-ray imaging and a large area electrophoretic display system for 
x-ray imaging is under development in contract CB-04341. A grant, CA-16543, 
is studying charge transfer electroradiography. Efforts to improve x-ray 
imaging and/or reduce x-ray exposure are subjects of several studies: contrast 
agents to improve CT scanning, grant CA-24879, and contract CB-84234; develop- 
ment of algorithms for x-ray dose reduction in CT scanning, contract CB-84235 
and grant CA-31217; image reconstruction from incomplete projections, grant CA- 
23818; computer analysis of bone tumor roentgenograms, grant CA-06263; study of 
optical quality effects on lung tumor detection, grant CA-23816; study of diag- 
nostic accuracy and x-ray image properties, grant CA-24625; development of a 
quick method of computing a single numbered dose index for CT scanning, grant 
CA-29I7I; and improved diagnostic quality of radiographic imaging for cancer 

198 



diagnosis, grant CA-24806. Improved imaging of breast tumors is the subject of 
several grants: CA-22803, computer evaluation of mammographic calcifications; 
CA-25372, new ' fluorescent x-ray tubes for mammography; CA-19622, low dose 
screening techniques for mammography; CA-19787, low dose breast electron radi- 
ography; and CA-26327, test phantoms for optimizing mammographic techniques. 
Other grants related to radiological imaging include CA-15882, construction of 
a transverse section x-ray camera; CA-23246, recognition of very small tumors 
in experimental animals; CA-24822, study of a 3-D tomogram viewer for cancer 
detection and therapy; CA-27823, development of a scanning equalization system 
for chest radiography; and CA-27875, post-mortem x-ray and histological com- 
parison of the sella turcica and pituitary glands. 

6. NON-RADIOLOGICAL IMAGING 

Imaging studies in the Cancer Diagnosis Research Program involving methods 
other than x-ray and radionuclides include use of proton and heavy ion beams, 
nuclear magnetic resonance (NMR) ultrasound, and thermography. Grant CA-27021 
supports a study of the imaging potential of proton, helium and carbon ion 
beams; extremely small tissue density differences are detected by ion beams 
but their lateral resolution is inferior to that of x-ray. Evaluation of a 
high magnetic field strength clinical NMR imaging spectrometer is being started 
under a new program project, grant CA-28881. Grant CA-15300 is developing NMR 
zeugmatographic imaging for cancer detection. Imaging with thermography at 
microwave and millimeter wave-lengths is being studied on grants CA-17642 and 
CA-28873. Several grants are investigating means to improve breast imaging 
with ultrasound: CA-I9019, ultrasound mammography; CA-24085, quantitative 
ultrasound imaging of the breast; CA-24257, breast tumor diagnosis by computed 
ultrasound; CA-25323, ultrasonic computed tomography of breast cancer; and 
CA-25634, development of an ultrasonic breast tissue phantom. Contrast agents 
for ultrasound diagnosis are being developed in contracts CB-84236 and CB-14337. 
An ultrasonic probe suitable for use in endoscopes is the goal of contract CB- 
74136. A scanning acoustic microprobe for cancer diagnosis by external imaging 
is under development and testing on grant CA-25938. Transpelvic ultrasonic 
evaluation of the prostate to improve early detection of prostate cancer is 
being studied on grant CA-27895. 

7. NUCLEAR MEDICINE 

Several grants are devoted to production of a variety of tagged tumor- 
seeking agents: CA-08349, CA-15787, CA-16861, CA-18153, CA-I9898, CA-24344, 
CA-26371, CA-26968, CA-28343, and CA-28561. Grant CA-27252 is carrying out 
studies of metabolism of bone scanning radiopharmaceuticals. Grant CA-24957 
and CA-22578 deal with the detection of malignant melanoma with radiopharma- 
ceuticals. Grant CA-22464 supports the study of gallium (III) and indium (III) 
chelates for the design of improved radiopharmaceuticals containing these 
metals. Program project grant CA-23417 is concerned with several basic studies 
to develop an improved nuclear imaging system for tumor detection. Grant CA- 
28105 involves development of an electronically collimated gamma tomography 
system for tumor imaging. 

8. MULTIPLE DISCIPLINES 

A number of contracts and one grant involve several disciplines and there- 
fore are presented in this last category of multiple disciplines. Contract 



199 



CB-84232 is studying diagnostic techniques including various imaging methods 
to improve pancreatic cancer diagnosis. Grant CA-25582 is studying fluores- 
cence bronchoscopy and photoradiation therapy with prophyrin derivatives. 
Contracts CB-74114 and CB-74212 are developing attachments for the colonoscope 
to facilitate its easier passage to the cecum. 

A collaborative study to determine the potential of x-ray imaging and cyto- 
loglcal examination of sputum in detection of early lung cancer is being con- 
ducted under contract CB-45007, CB-45037, and CB-53886. A fourth related 
contract, CB-43868, is concerned with the data management of the collaborative 
lung study. The longitudinal study, began in 1974, involves 30,000 men and 
will require several more years of follow-up before definitive results will be 
available. 

A large-scale longitudinal colon cancer screening study using the Hemoccult 
test for human blood in the feces is the task of contract CB-53862. Several 
more years of follow-up on the 45,000 subjects will be required to define the 
usefulness of the screening procedure in detecting early colon cancer. 



200 



BIOCHEMISTRY 



ROl-CA-14025 Multiple Forms of HGH: Measurements and Actions 

Willard Vander Laan Scripps Clinic and Research Foundation 

ROl-CA-14185 Modified Nucleosides in Cancer and Normal Urines 
Girish Chheda Roswell Park Memorial Institute 

ROl-CA-14335 EPR Studies on Detection and Treatment of Cancer 
John D. Zimbrick University of Kansas Lawrence 

ROl-CA-19606 Serum Urine and CSF RNases in Health and Disease 
Charles A. Dekker University of California Berkeley 

ROl-CA-19634 Serum Lipoproteins in Patients with Breast Cancer 
Frederick Aladjem University of Southern California 

ROl-CA-21958 Cancer Detection by Serum Analysis of Heteroglycans 
James D. Morre Purdue University 

ROl-CA-22065 Lymphoma: Diagnosis by Blood and Liver Tests 
Annemarie Herzfeld New England Deaconess Hospital 

ROl-CA-22599 Programs of Normal and Malignant Lymphocytes 

Allen Silverstone Sloan-Kettering Institute for Cancer Research 

ROl-CA-22885 Lung Cancers and Carcinomatous Neuromyopathies 
Koichi Ishikawa University of Southern California 

ROl-CA-23848 Steroid Uptake in Hormonally Dependent Cancers 
George H. Barrows University of Louisville 

ROl-CA-23945 Assessment of Malignancy in Human Chondrosarcomas 

Lawrence C Rosenberg Montefiore Hospital and Medical Center 

ROl-CA-25016 Human Lung Neoplasms: Applications of Enzymes-Pathology 
Olga Greengard Mount Sinai School of Medicine 

ROl-CA-25210 Origins of Urinary Nucleosides in Tumor Tissue 

Ernest Borek AMC Cancer Research Center and Hospital 

ROl-CA-25357 Cyclic GMP Levels in Gynecologic Neoplasms 
Chandralekha Duttagupta Yeshiva University 

ROl-CA-25376 Development of Serum Nuclease Isozyme Test for Cancer 
K. C. Tsou University of Pennsylvania 

ROl-CA-25548 Isozymes of Human Tumor Cells In Vitro and In Vivo 

Jorgen Fogh Sloan-Kettering Institute for Cancer Research 

ROl-CA-26652 Histaminase as a Biochemical Marker for Human Cancer 
Chi-Wei Lin Massachusetts General Hospital 



201 



ROl-CA-29062 Vasoactive Intestinal Peptide, Leukocytes, Leukemia 

Mary O'Dorisio Ohio State University Research Foundation 

ROl-CA-29056 Large Bowel Cancer and Colonic Microbial Metabolism 
David Karlin University of Texas 

ROl-CA-30667 A Study of Cancer Associated Colonic Mucin 

Clement R. Boland Veterans Administration Medical Center 

ROl-CA-31187 Biochemistry and Clinical Application of Acid Phosphatase 5 
Kwok-Wai Lam Albany Medical College 

ROl-CA-31218 Thyrotropins from Tumors of Trophoblastic Origin 
Syed M. Amir Beth Israel Hospital 



2. IMMUNODIAGNOSIS 



POl-CA-04486 Pathology of the Digestive Tract Mucous Membrane 
Norman Zamcheck Boston City Hospital 

ROl-CA-17742 Radiological Localization of Human Tumors 

David M. Goldenberg University of Kentucky Medical Center 

ROl-CA-20286 Breast Neoplasia Diagnosis with Specific Antibodies 

Roberto L. Ceriani Children's Hospital Medical Center Northern 

California 

ROl-CA-22137 Calcitonin as a Marker for Cancer of the Breast 
Omega L. Silva Howard University 

ROl-CA-22141 Protein SAA in Neoplastic Disease 

Merrill D. Benson Indiana University School of Medicine 

ROl-CA-22595 Detection of Medullary Thyroid Cancer in Families 
Charles E. Jackson Henry Ford Hospital 

ROl-CA-23382 Investigation of Human B-Lipotropin 

Eckehart Wiedemann University of California Berkeley 

R23-CA-23990 Immunodiagnosis of Prostatic Cancer 

Ching-Li Lee Roswell Park Memorial Institute 

ROl-CA-24376 Immunological Heterogeneity of CEA 
James F. Primus University of Kentucky 

ROl-CA-24645 Significance of GP48 in Diagnosis of Breast Cancer 
Zoltan A. Tokes University of Southern California 

ROl-CA-25338 Thyroglobulin Radioimmunoassay in Patients with Thyroid Cancer 
Merl A. Charles University of California at Irvine 



202 



ROl-CA-25381 Tyrosinase as a Marker for Human Malignant Melanoma 

Kenji Nishioka University of Texas System Cancer Center 

ROl-CA-25584 Clinical CEA-Tumor Radioimmunodetection 
David M. Goldenberg University of Kentucky 

ROl-CA-26246 Assay of Human Tumor or Organ-Associated Antigens 
Calvin A. Saravis Boston City Hospital 

ROl-CA-26948 Nuclear Antigens as Markers in Hematopoiesis 
Robert C. Briggs Vanderbilt University 

ROl-CA-27081 Immunodiagnosis of Mesothelioma 

Gurmukh Singh University of Pittsburgh 

ROl-CA-27213 Detection of Urogenital Normal and Neoplastic Antigens 
Robert W. Green Duke University Medical Center 

ROl-CA-28305 Possible Systemic Sequals for Tumor 

Earl M. Ritzi University of Tennessee Center for Health Sciences 

ROl-CA-28462 Radiolabeling of Tumor Antibodies 

William C. Eckelman George Washington University 

ROl-CA-29211 Immunohistologic Study of Uterine Cancer 

Clive R. Taylor University of Southern California School of 

Medicine 

ROl-CA-29225 Clinical Application of Esterase, a Monocyte Marker 
Kwok-Wai Lam Albany Medical College 

ROl-CA-29552 Differential Counting of Lymphocyte Subpopulations 
Marius Teodorescu University of Illinois Medical Center 

ROl-CA-29639 Tumor Imaging with Radiolabeled Monoclonal Antibody 
Steven M. Larson VA Medical Center 

ROl-CA-30019 Purification of Tumor Antigens of Defined Specificities 
Risab K. Gupta UCLA Center for Health Sciences 

ROl-CA-30255 Immunolocalization of Human Malignant Tumors 
Elliot Alpert Baylor College of Medicine 

ROl-CA-31762 Immunologic Diagnosis of Myeloblastic Leukemia 

Robert N. Taub Columbia University School of Medicine 



3 . CYTOLOGY 



ROl-CA-13271 Automated Cancer Cell Diagnosis by the TICAS Method 
George Wied University of Chicago 



203 



ROl-CA-17353 Marrow Culture Studies in Human Myeloid Leukemias M 

Malcolm A. Moore Sloan-Kettering Institute for Cancer Research ^ 

ROl-CA-19552 New Fluorescent Markers for Cancer Diagnosis 

Kwan C. Tsou University of Pennsylvania 

ROl-CA-22942 Differentiation of Cultured Leukemic Cells 

Sandra R. Wolman New York University 

ROl-CA-23393 Flow Analysis of Human Malignant Lymphoid Cells 

Raul C. Bray Ian University of Florida 

ROl-CA-24466 Ultrafast Scanner Microscope in Laboratory Automation 

Roland V. Shack University of Arizona 

ROl-CA-24932 Microspectrophotometric Nuclear DNA Study of Gynecologic Cancers 

Yao S. Fu Case Western Reserve University 

ROl-CA-25348 Flow Cytometry/Autoradiography Monitoring of Leukemia 

Michael Andreeff Sloan-Kettering Institute for Cancer Research 

ROl-CA-26863 Identification of CEA in Cytology Specimens 

Robert R. Pascal St. Luke ' s-Roosevelt Institute for Health Sciences 

ROl-CA-27123 Application of Fluorescent Probes to Clinical Cancer 

Jay E. Valinsky Rockefeller University 

ROl-CA-27283 Early Detection of Transformed Cells 

Susan P. Hawkes Michigan Molecular Institute 

ROl-CA-27313 A Search for Preneoplastic Cell Markers in Sputum 

Stanley D. Greenberg Baylor College of Medicine 

ROl-CA-28704 Chromatin Probes for Distinguishing Malignant Cells 

Zbigniew Darzynkiewicz Sloan-Kettering Institute for Cancer Research 

ROl-CA-28706 Cell Positioning System: Development and Use in Cancer 

Harry W. Tyrer Cancer Research Center 

ROl-CA-28724 Biophysical Probes for Malignant Cells 

Paul Todd Pennsylvania State University 

ROl-CA-28770 Biophysical Probes for Automated Cytology 

Kwan C. Tsou University of Pennsylvania 

ROl-CA-28771 Cytology Automation 

Barthel Barlogie M.D. Anderson Hospital and Tumor Institute 

ROl-CA-28833 Cytologic Characterization of Rat Urothelium 

Ian T. Young Lawrence Livermore Laboratory 

ROl-CA-28886 Indexed Cell Sorting 

Phillip Dean Lawrence Livermore Laboratory 

204 



ROl-CA-28921 Merocyanine Dyes as Leukemia-Specific Probes 
Robert A. Schlegel Pennsylvania State University 

R13-CA-29859 Eighth Conference on Analytical Cytology 

Myron R. Melamed Memorial Sloan-Kettering Institute for Cancer 

Research 

ROl-CA-30148 Development of Lanthanide Fluorescent Stains 
Lidia M. Vallarino Virginia Commonwealth University 

ROl-CA-30582 Multistage Slit-Scan Prescreening System 

Leon L. Wheeless University of Rochester Medical Center 



PATHOLOGY 



ROl-CA-21905 Pituitary Adenomas: Structure-Function Relations 
Calvin Ezrin Cedars-Sinai Medical Center 

ROl-CA-22101 Study of Head and Neck Cancer by Serial Section 
John A. Kirchner Yale University 

ROl-CA-23922 Breast Cancer 

Thomas G. Pretlow University of Alabama in Birmingham 

ROl-CA-26422 Clinico-Biologic Correlation in Lymphoma and Leukemia 
Henry Rappaport City of Hope National Medical Center 



5. RADIOLOGICAL IMAGING 



ROl-CA-15882 Transverse Section X-Ray Camera 

Gordon L. Brownell Massachusetts General Hospital 

ROl-CA-16543 Investigation of Charge Transfer Electroradiography 
Ivor Brodie SRI International 

ROl-CA-19787 Low Dose Breast Electron Radiography (Tonography) 
Gary S. Shaber Thomas Jefferson University 

ROl-CA-23246 Recognition of Very Small Tumors 
Tamas Sandor Harvard University 

ROl-CA-23816 Optical Quality Effects on Lung Tumor Detection 
Robert D. Moseley, Jr. University of New Mexico 

R23-CA-23818 Image Reconstruction from Incomplete Projections 

Robert M. Lewitt State University of New York at Buffalo 

ROl-CA-24625 Diagnostic Accuracy and X-Ray Image Properties 
George Revesz Temple University 



205 



ROl-CA-24806 Radiographic Imaging for Cancer Diagnosis 
Kunio Doi University of Chicago 

ROl-CA-24822 3-D Tomogram Viewer for Cancer Detection and Therapy- 
Brent S. Baxter University of Utah 

R23-CA-24879 Malignant Perfusion: Applied to Diagnosis and Therapy 
Stuart Young Stanford University School of Medicine 

R23-CA-25372 New Fluorescent X-Ray Tubes for Mammography 
G. Allan Johnson Duke University 

ROl-CA-26327 Test Phantoms for Optimizing Mammographic Techniques 
Leonard Stanton Hahnemann Medical College and Hospital 

ROl-CA-27823 A Scanning Equalization System for Chest Radiography 
Donald B. Plewes University of Rochester 

ROl-CA-27875 Post-Mortem X-Ray and Histology of Sella and Pituitary 
Dennis D. Spencer Yale University 

ROl-CA-29171 Integral and Mean Dose in CT Examinations 

Feargus O'Foghludha Duke University School of Medicine 

ROl-CA-31217 New Reconstruction Algorithms for Computed Tomography 
Robert M. Lewitt University of Pennsylvania 



6. NON-RADIOLOGICAL IMAGING 



ROl-CA-15300 Application of NMR Zeugmatography in Cancer Research 

Paul C. Lauterbur State University of New York at Stoney Brook 

RO1-CA-19019 Ultrasound Mammography 

Gilbert Baum Yeshiva University - Albert Einstein College of 

Medicine 

RO1-CA-24085 Breast Diagnosis Quantitative Imaging by Ultrasound 
James F. Greenleaf Mayo Foundation 

ROl-CA-24257 Breast Tumor Diagnosis by Computed Ultrasound 
William Swindell University of Arizona 

ROl-CA-25323 Ultrasonic Computed Tomography of Breast Cancer 

Paul L. Carson University of Colorado Medical Center 

ROl-CA-25634 Development of Ultrasonic Breast Tissue Phantom 
James A. Zagzebski University of Wisconsin 

ROl-CA-25938 Scanning Acoustic Microprobe for Cancer Diagnosis 
Frank E. Barber Harvard University 



206 



ROl-CA-27021 Heavy-Ion Radiography and Cancer 

Cornelius A. Tobias University of California Berkeley 

ROl-CA-27895 Transpelvic Ultrasound Evaluation of the Prostate 
Aubrey M. Palestrant Beth Israel Hospital 

ROl-CA-28873 Non-invasive Sensing of Subcutaneous Temperatures 
Alan H. Barrett Massachusetts Institute of Technology 

POl-CA-28881 Evaluation of High Field Medical NMR Imaging 
Sadek K. Hilal Columbia University 



7. NUCLEAR MEDICINE 



ROl-CA-08349 Potential Tumor or Organ Imaging Agents 

Raymond E. Counsell University of Michigan at Ann Arbor 

ROl-CA-16861 Bifunctional Chelating Agents in Tumor Localization 
Claude F. Meares University of California Davis 

ROl-CA-17742 Radiological Localization of Human Tumors 
James F. Primus University of Kentucky 

ROl-CA-18153 Cancer Studies with New Radioactive Scanning Compounds 

John S. Laughlin Sloan-Kettering Institute for Cancer Research 

ROl-CA-19898 . Synthesis of New '^Se Organ Specific Imaging Agents 
Michael A. Davis Northeastern University 

ROl-CA-22464 Gallium (III) Complexes in Aqueous Solution 
Arthur E. Martel Texas A & M University 

ROl-CA-22578 Melanoma Delineating Radiopharmaceuticals 

Ned D. Heindel Hahnemann Medical College and Hospital 

POl-CA-23417 Improved Tumor Imaging in Nuclear Medicine 
Harrison H. Barrett University of Arizona 

R23-CA-24344 Synthesis, Chemistry of Technetium Radiopharmaceuticals 
Michael J. Clarke Boston College 

ROl-CA-24597 Melanoma Detection with Radiopharmaceuticals 
Harold L. Atkins Brookhaven National Laboratory 

ROl-CA-26371 Radiopharmaceuticals for Positron Tomography in Cancer 
David R. Elmaleh Massachusetts General Hospital 

ROl-CA-26968 Improved Tumor Imaging with New Radioactive Liposomes 

Donald J. Hnatowich University of Massachusetts Medical School 



207 



ROl-CA-27252 Metabolism of Bone Scanning Radiopharmaceuticals 
Charles D. Russell University of Alabama in Birmingham 

ROl-CA-28105 An Electronically Collimated Gamma Tomography System 
Manbir Singh University of Southern California 

ROl-CA-28343 Bifunctional Chelates in Cancer Imaging and Therapy 
David A. Goodwin V.A. Medical Center 

ROl-CA-28561 Development of Tumor and Organ Imaging Agents 

Larry A. Spitznagle University of Maryland at Baltimore 



8. MULTIPLE DISCIPLINES 



'ROl-CA-25582 Fluorescence Endoscopy and Photoradiation Therapy 
Oscar J. Balchum University of Southern California 



208 



CONTRACT RESEARCH SUMMARY 

Title: Contrast Agents in Detecting Liver Metastases with Computerized X-ray 
Tomography 

Principal Investigator: Dr. Michael A. Davis 

Performing Organization: Brigham & Women's Hospital 

City and State: Boston, MA 

Contract Number: NOl-CB-84234 

Starting Date: 7/01/78 Expiration Date: 6/30/81 

Goal: To improve detection of small metastatic lesions in the liver by improved 
use of contrast agents to provide appropriate image enhancement in x-ray compu- 
terized tomographic studies. 

Approach: The work, which will be carried out over a three-year period, will con- 
sist of six tasks: synthesis of new hepatic-specific contrast agents; preliminary 
evaluation in animal models; toxicity evaluation in animals; pilot clinical stud- 
ies with currently available contrast agents; pilot clinical studies of promising 
new agents with acceptable toxicity characteristics; and clinical trials on the 
most promising contrast agent. 

Progress: Work has progressed in two major areas: 

1) Non-metabolizable reticuloendothelial (RE) tissue imaging agents. 

Acute toxicity trials of cerium oxide indicate that there is no acute toxicity 
at doses anticipated to provide 50 Hounsfield Units (HU) of contrast enhancement 
of the normal liver. Extensive sub-acute toxicity testing is under way, prior to 
human clinical trials. 

2) Development and testing of metabolizable RE tissue agents. 

A. Radiopaque liposomes - Two varieties of radiopaque liposomes (phospho- 
lipid vesicles) were created. Those made from egg yolk lecithin, cholesterol, and 
stearylamine in a 4:1:1 molar ratio and carrying meglumine and sodium diatrizoate 
demonstrated prolonged blood pool opacification and clearance through reticuloen- 
dothelial tissues. Contrast enhancement of the heart, vessels, liver, and spleen 
was recognized. Contrast enhancement of these tissues was prolonged. At 10 min- 
utes, nearly 30% of the injected dose was still present in the blood pool, only 
falling to 23% at 60 minutes. Maximal measured contrast enhancement of the blood 
was 116 HU, the liver enhanced 62 HU, and the spleen 87 HU, following a radiopaque 
liposome dose that contained 190 mg iodine. These peak levels were nearly sus- 
tained for 60 minutes and slowly decreased to base line levels over one day. 
There was strikingly greater opacification of all of these structures than pro- 
vided by an equivalent amount of iodine in Renografin. 

Additional studies on materials prepared from soy bean lecithin, cholesterol, 
and stearylamine in 8:1:1 molar ratio showed that this material went predominantly 
to the spleen. Peak spleen numbers were consistently greater than 100 HU follow- 
ing doses of 33:3 mg I in radiopaque liposomes. 

B. Radiopaque albumin - Synthesis of radiopaque, iodinated albumin has 
been accomplished. This material is currently being tested in animals to assess 
its biodistribution. 

Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: ^293, 000 

209 



CONTRACT RESEARCH SUMMARY 

Title: Digital Image Processing Techniques in Cytology Automation 

Principal Investigator: Dr. Kenneth R. Castleman 

Performing Organization: JPL-Calif ornia Institute of 

Technology 
City and State: Pasadena, CA 

Contract Number: YOl-CB-70314 

Starting Date: 9/19/77 Expiration Date: 9/30/81 

Goal: Identification of the best performing image analysis algorithms and 
estimations of the expected performance and cost per specimen of an automated 
cervical cancer pre-screening system using these algorithms. 

Approach: The contractor shall determine whether or not the current state of 
the art in pattern recognition is adequate to support the development of an 
economically viable cytological screening instrument based on single cell clas- 
sification. Specimen preparation protocol will be optimized so as to produce 
acceptable single cells for digitization. A large number of digitized images 
will be accumulated in a library for a subsequent feature extraction program 
to produce a feature data base. A series of classification experiments will 
be run on the feature data base to select the subset of cell measurements that 
yield the best overall performance. An analysis will produce estimates of the 
cost and performance using these techniques in practice. 

Progress: A new specimen preparation protocol has been developed. Over 10,000 
digitized cell images were acquired and classification algorithms were devel- 
oped and tested. New statistical methods for optimal specimen classification 
and analysis have been developed. The analysis of a cascade of a cell classi- 
fier followed by a specimen classifier has provided quantitative criteria for 
evaluating all classifier effectiveness as part of an overall system. This 
analysis has also provided guidelines for tuning the parameters of the cell 
classifier. A cost/utility analysis has indicated that the cell classification 
algorithms chosen can operate with error rates that are adequate for routine 
screening. 

The second phase of the study has been initiated. This phase will evaluate 
the performance of the cell classification algorithms on cells selected and 
segmented without human intervention. New algorithms for fully automatic cell 
finding and segmentation have been developed, and are undergoing test and 
evaluation. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



210 



CONTRACT RESEARCH SUMMARY 

Title: Diagnostic Application of Monocyte Function in Cancer 

Principal Investigator: Dr. Ralph Snyderman 

Performing Organization: Duke University Medical Center 

City and State: Durham, NC 

Contract Number: NOl-CB-74121 

Starting Date: 9/30/77 Expiration Date: 6/29/81 

Goal: To determine the significance of abnormal monocyte functions in humans 
with cancer. 

Approach: The relationship between the clinical status of cancer patients and 
their monocyte function in vitro is being determined. Parameters of phagocytic 
cell function such as chemotactic responsiveness, phagocytosis, superoxide pro- 
duction and chemotactic factor binding are being examined. We seek to deter- 
mine the significance of the depression of monocyte chemotaxis which has been 
found in cancer patients. The effect of tumor removal in phagocytic cell func- 
tion is being examined. The effects of effusions from patients with various 
types of cancer on the function of normal monocytes are also being examined. 

Progress: We have previously demonstrated that monocyte chemotactic respon- 
siveness in vitro is depressed in approximately 60% of patients with cancer. 
Removal of the tumor by surgery or by immunotherapy reverses this defect, sug- 
gesting that tumor associated factors might be responsible. Using a newly 
developed assay for measuring one of the early responses of monocytes to chemo- 
tactic stimuli, i.e., the change in shape from round to an elongated, polarized 
configuration, we are now examining fluids from patients for the presence of 
soluble inhibitors of chemotactic responses. We have found inhibitory activity 
for monocyte polarization in the fluids of all (22/22) of the cancer patients 
studied thus far, representing 15 different tumor types, but in none of 17 
fluids from patients with non-malignant diseases. Fractionation of fluids by 
high pressure liquid chromatography revealed three peaks of inhibitory activ- 
ity: > 200,000 daltons; 46,000 + 13,000 daltons; and 21,000 + 3,000 daltons. 
The inhibitory activity was shown to be heat stable (56°C, 30 min) and trypsin- 
sensitive. Three different monoclonal antibodies to the P15(E) structural com- 
ponent of murine type C retroviruses were capable of absorbing the inhibitory 
activity from all eight fluids tested while six other monoclonal antibodies had 
no effect. We are currently in the process of further characterizing this 
inhibitory material and screening for its presence in the serum of cancer 
patients. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



211 



CONTRACT RESEARCH SUMMARY 

Title: Screening for Medullary Carcinoma of the Thyroid 

Principal Investigator: Dr. Samuel A. Wells, Jr. 

Performing Organization: Duke University Medical Center 

City and State: Durham, NC 

Contract Number: NOl-CB-63994 

Starting Date: 6/15/76 Expiration Date: 6/14/81 

Goal: To establish a periodic screening program of relatives of patients with 
medullary thyroid carcinoma (MTC) • « 

il 
Approach: A cohort of patients comprised of seven kindreds in which MTC is 
proven histologically will be established and be large enough to offer a sta- 
tistically significant number of family members. Those family members will be ' 
screened for serum calcitonin levels by radioimmunoassay methods in order to 
detect MTC in its earliest stage of development. A long-term follow-up program 
will be conducted in these families to determine the natural history of the 
disease and the effectiveness of surgical and possible chemical therapy. -.^ 

Progress: Our study of approximately 800 subjects from 13 kindreds with multi- 
ple endocrine neoplasia, type II (medullary thyroid carcinoma (MTC), pheochro- 
mocytoma and hyperparathyroidism) has completed its fifth year. We have demon- 
strated that the combined infusion of calcium gluconate (2 mg/kg/1 min) 
followed by pentagastrin (0.5 /xg/kg/5 sec) is the most sensitive method of ^ 
stimulating calcitonin (CT) release from MTC cells. This combined infusion ^ 
serves as the most efficient method for establishing the early diagnosis of | 
MTC. Over the last year we have identified two kindreds who inherit only MTC [ 
with none of the extrathyroidal manifestations of multiple endocrine neoplasia 
(MEN), types Ila or lib. Furthermore, the MTC in these patients appears to be ! 
less aggressive than the MTC occurring in subjects with MEN Ila or subjects 
with MEN lib. 

Also, over the last year, Dr. Stephen Baylin and associates at the Johns 
Hopkins Hospital have utilized immunoperoxidase techniques for determining 
calcitonin content in primary MTC. It has been learned that patients whose 
tumors stain heterogenously with calcitonin have a relatively poor prognosis 
compared to those whose tumors demonstrate homogenous calcitonin staining. We 
are conducting further experiments to define the utility of this prognostic 
indicator. 

We continue to evaluate chemotherapeutic agents in the treatment of 
patients with metastatic MTC. Thus far, of several chemotherapeutic agents, 
none has proven of benefit with the possible exception of adriamycin in two 
patients. 



Project Officer: Mr. Louis P. Greenberg 
Program: Diagnosis 
FY 81 Funds: 



212 



CONTRACT RESEARCH SUMMARY 



Title: Data Management System for NCI Serum Panels 



Principal Investigators: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-74210 
8/08/77 



Dr. Lee A. Richman, Dr. C. M. Dayton 
Ebon Research Systems 
Washington, DC 



Expiration Date: 8/07/82 



Goal: To provide data management and statistical programming support for 
research projects being conducted by the Diagnosis Program in order to deter- 
mine which serum tests are best able to detect early stages of cancer. 

Approach: To perform statistical analyses of data from NCI serum panel evalua- 
tions and to prepare summary reports of the results. 

Progress: Approximately two serum panels have been forwarded to us each month, 
and we have conducted the following statistical analyses on each: 

(1) The production of plots, such as histograms, so the distributions 
across clinical groups can be compared. In most instances an analysis of vari- 
ance accompanies each plot. 

(2) Chi-square (2 x 2) tables for a variety of clinical comparisons. 
Accompanying each chi-square value are the associated two-tailed probability, 
sensitivity, specificity, and number of misclassif ications for the comparison. 
When cut-scores separating positive from negative groups are found by inspec- 
tion of the data — minimizing the number of misclassif ications — a Kolmogorov- 
Smirnov statistic is used instead of chi-square. 

(3) McNemar chi-square tests, when appropriate, to determine whether the 
rates of positive classifications are the same for two assays. 

(4) Discriminant analyses for multiple comparisons to combine information 
from the assays into a single prediction function. 

(5) Gail-Green procedures for locating optimal cutting scores. 

(6) Scattergrams for pairs of assays. 

In general, none of the panels analyzed since September 1, 1980 (including 
a large comparative analysis of assays of immune complexes) were adequate in 
their ability to discriminate between cancer and non-cancer cases. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: $76,000 



213 



CONTRACT RESEARCH SUMMARY 

( 

Title: Immunodiagnostic Markers for Breast Cancer 

Principal Investigator: Dr. Rajender K. Chawla 

Performing Organization: Emory University- 

City and State: Atlanta, GA 

Contract Number: NOl-CB-84308 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal: To evaluate urinary/plasma levels of EDCl, the principal component of 
cancer-related proteinuria, as an immunodiagnostic marker for breast cancer. 

Approach: Pure EDCl and inter-a-trypsin inhibitor (lATI) and high titer anti- 
serum to EDCl will be prepared. Standardized immunologic methods (e.g., radio- 
immunoassay (RIA) or immunoelectrophoretic techniques) will be developed to 
monitor EDCl in urine/plasma of i) normal healthy women; ii) women with meta- 
static breast cancer; iii) patients with non-neoplastic diseases; and iv) pre- 
operative patients with localized breast mass. A postoperative longitudinal 
study will monitor EDCl levels of patients in group iv with malignant lesions. 

Progress: A novel glycoprotein EDCl, MW 27.5 K, isolated originally from urine 
of a leukemic patient was found to be immunologically related to a normal plas- 
ma protein, lATI (MW 170 K). Urinary EDCl levels have been measured in the 
above four groups of subjects. The ave + SEM values (mg/g urinary creatinine) 
were as follows: i) normal women: 8.0 + 2.2 and ii) metastatic breast cancer 
patients: 98.6 + 11. Sixty-six out of 82 patients with noncancer diseases had 
an ave EDCl level of 14.6 + 4; the remaining had an ave of 94.8+1^* Among 
the latter subgroup were patients with renal failure, rheumatoid arthritis, and 
infectious diseases. The immunoreactive material excreted in these non-cancer 
diseases was of higher MW (presumably lATI) and was positively correlated with 
the degree of renal insufficiency. In the 26 preoperative patients, subse- 
quently shown to have benign lesion, the average EDCl excretion was 21.5 + 3.4; 
24 of these patients were in the normal (<15 mg) to light (15-30 mg) range and 
2 were in the Intermediate (31-45 mg) range. In the other preoperative sub- 
group (25 patients) subsequently shown to have malignant lesions, the ave EDCl 
excretion was 43.1 + 7.6 mg, with 8 normal, 5 light, 4 intermediate and 8 heavy 
(>45 mg) excretors. Clinical evaluation of preoperative patients with malig- 
nant lesions and its correlation with EDCl levels showed a direct correlation 
between the number of nodes and the level of EDCl. Postoperative follow-up in 
the heavy excretor preoperative patients showed a marked decline in EDCl fol- 
lowing removal of the tumor (from 171 to 21 mg). Analyses of fractions obtained 
by gel filtration of plasma of metastatic breast cancer patients showed no sig- 
nificant accumulation of EDCl, suggesting rapid clearance of EDCl from plasma. 
Preliminary data indicate that total IR-IATI in breast cancer is about 2/3 of 
that in the normals. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: Ml, 783 



214 



• 



CONTRACT RESEARCH SUMMARY 

Title: Clinical Evaluation of Immunodiagnostic Tests for Cancer 

Principal Investigator: Dr. Adly N. Ibrahim 

Performing Organization: Georgia State University 

City and State: Atlanta, GA 

Contract Number: NOl-CB-84267 

Starting Date: 2/01/78 Expiration Date: 6/30/81 

Goal: Immunodiagnosis of cervical and head and neck cancers. 

Approach: Antisera are prepared in rabbits against cervical cancer tissues. 
Elimination of antibodies against normal tissues and normal human sera is 
carried out by various methods. These antisera will be used for the detection 
of circulating tumor-associated antigens (C-TAA) in sera of patients with cer- 
vical and head and neck cancers. Serum controls including those from normal 
individuals and from patients with benign diseases of these sites will be used. 

Progress: The contractor has purified hyperimmune sera against partially puri- 
fied cervical cancer tumor-associated antigens (CaCx TAA) using ion exchange 
and affinity chromatography. These antisera were evaluated for the detection 
of C-TAA in sera from cervical cancer patients using an indirect method of 
enzyme-linked immunosorbent assay (ELISA) as well as the immunodiffusion tech- 
nique (ID). ELISA has increased significantly the detection of C-TAA. Specif- 
icity was high though not absolute. Efforts will continue to increase the sen- 
sitivity and specificity of ELISA. Preparation of monoclonal antibodies 
against CaCx TAA's has been initiated using hybridomas. Such antibodies will 
be used for (1) immunodiagnosis of cervical and head and neck cancers using 
ELISA and RIA. It is expected that such antibodies will increase the specific- 
ity of the tests, and (2) to study the TAA fractions obtained after purifica- 
tion of CaCx TAA's. Antisera prepared against a continuous cell line derived 
from cervical cancer tissues using immunodiffusion adsorption-in-gel technique. 
Using coded human sera in blind tests, anti-C4lI sera detected C-TAA in 24/36 
(66.6%) sera from patients with cervical cancer. Five of 126 (3.9%) control 
sera gave false positive reactions. This continuous cell will also be used to 
purify C4II TAA's, and to prepare monoclonal antibodies. Preliminary results 
indicate that CaCx TAA's and C4II TAA's may be identical. Studies indicated 
the presence of at least 3 CaCx TAA's. Purification, isolation and characteri- 
zation of each TAA are pursued actively. During the progress of the hybridoma 
work, hyperimmune mouse sera and ascitic fluids against CaCx TAA's were evalu- 
ated for diagnosis of cervical cancer using ELISA. Results indicated that the 
mouse antibodies are superior and more specific than those of rabbits for this 
purpose. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: $20,835 



215 



CONTRACT RESEARCH SUMMARY 

Title: Immunologic Markers Applicable to Cytology Automation 

Principal Investigator: Dr. Laure Aurelian 

Performing Organization: Johns Hopkins University 

City and State: Baltimore, MD 

Contract Number: NOl-CB-74170 

Starting Date: 7/18/77 Expiration Date: 7/27/81 

Goal: To study qualitative and quantitative antigenic changes in premalignant 
and malignant cells. 

Approach: The contractor shall isolate HSV antigens: (i) total HSV-2 anti- 
gens, (ii) total HSV-1 antigens, (iii) AG-e, (iv) AG-4/ICP 10 and prepare cor- 
responding purified antisera. The contractor shall also attempt to obtain 
antiserum to VP143 from an independent investigator. The sera will be used to 
stain hximan gynecologic specimens in indirect immunofluorescence. The sensi- 
tivity and specificity of these antisera in terms of their ability to discrim- 
inate premalignant and malignant cells from normal cells will be determined. 

Progress: AG-e purified by crossed Immunoelectrophoresis has been resolved by 
SDS acrylamide gel electrophoresis into two component proteins designated ICP 
12 (MW 140,000) and ICP 14 (MW 130,000). Antisera were prepared against AG-e, 
ICP 12 and ICP 14 and their reactivity determined. ICP 12 and ICP 14 appear 
to be virion envelope proteins. 

In indirect immunofluorescence, antisera to AG-e, ICP 12 and ICP 14 stain 
exfoliated atypical but not normal cells. In a double blind study designed to 
compare immunofluorescent markings of specimens by antiserum to AG-e, a very 
good correlation (80-93.8%) was observed between staining with anti-AG-e serum 
and routine manual screening. Very good correlation is observed in ongoing 
blind studies (including specimen by specimen and cell by cell comparisons) 
between the staining potential (frequency of positive patients and percent of 
positive cells/patients) of antisera to total HSV-2 antigens, to total HSV-1 
antigens and to AG-e. However, antisera to ICP 12 and ICP 14 stain a smaller 
number of cells and a lower frequency of patients are responsive. Presently 
ongoing studies indicate that reactivity with antiserum against AG-4/ICP 10 
reflects the progression of the cervical lesion. Thus 4/10 patients with mild 
dysplasia, 7/10 with moderate dysplasia and all those with a lesion diagnosed 
as, at least, marked dysplasia had AG-4/ICP 10 positive cells. Normal cells 
from these patients and normal control women were negative. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



216 



CONTRACT RESEARCH SUMMARY 



Title: Lung Cancer Control: Detection and Treatment 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CN-45037 
1/01/73 



Dr. John K. Frost 

Johns Hopkins University 

Baltimore, MD 



Expiration Date: 7/15/81 



Goal: To determine the effect on mortality from lung cancer of early detection 
of this disease by sputum cytology followed by surgical removal of the tumor. 

Approach: The study population consists of males who, at the time of enroll- 
ment, were 45 years or older and had smoked a pack or more of cigarettes a day. 
Individuals responding to invitation, by direct mailing and other means, were 
randomly assigned to one of two groups: the "X" group receives an annual chest 
x-ray only; the "CX" group receives an annual chest x-ray, an annual sputum 
induction plus a sputxim cytologic examination every four months. Subjects sus- 
picious or positive for cancer are carefully evaluated, including localization 
of the cancer by fiberoptic bronchoscopy. Cancers are removed surgically when 
possible. All subjects will be followed for at least 5 years after final 
screening to determine actuarial survival. 

Progress: A total of 10,387 men have been enrolled in this project, of which 
5,161 were randomized into the "X" group and 5,226 into the "CX" group. To 
date, 740 have died, while 1,669 have withdrawn from the study or moved from 
the area. Currently 7,978 remain as active participants, 4,025 in the "X" 
group and 3,953 in the "CX" group. At the initial screening 78 cancers were 
detected (40 in the X group and 38 in the CX group) for a "detected prevalence" 
of 7.5 per 1,000. Following an initial negative screening, a total of 200 can- 
cers appeared (102 in the X group and 98 in the CX group) for an "annual inci- 
dence" of 4.7 per 1,000. In the CX group, 37 (65%) incidence cases were 
detected at AJC Stage I (localized), 10 (27%) of these were in situ and 27 
(73%) invasive; 41 additional cases occurred between screenings, 15% at Stage 
I. In the X group, 32 (51%) incidence cases were detected at AJC Stage I, all 
invasive; 39 additional cases occurred between screenings, 21% at Stage I. 
Among the 98 cases occurring in the CX group after initial screening, 46 (47%) 
were resected; 45 (44%) of the 102 incidence cases occurring in the X group 
were resected. In the screened population, 278 total lung cancers have 
appeared: 142 in the X group and 136 in the CX group. Among the 142 cases of 
lung cancer found in the X group, 86 (61%) have died; of the 136 lung cancer 
cases in the CX group, 66 (49%) have died. Lung cancer mortality has not yet 
been shown to be significantly different between the CX (30 per 10,000 person- 
years) and X (41 per 10,000 person-years) groups, nor has a significant differ- 
ence been shown yet from that expected from application of the age and smoking 
specific lung cancer mortality rates of historical control groups. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $760,000 



217 



CONTRACT RESEARCH SUMMARY 

Title: Immune Assays for Enzymes and Isozymes in Cancer 

Principal Investigator: Dr. Joel H. Shaper 

Performing Organization: Johns Hopkins University 

City and State: Baltimore, MD 

Contract Number: NOl-CB-84260 

Starting Date: 9/22/78 Expiration Date: 9/21/81 

Goal: Development of immunodiagnostic assays for cancer using an enzyme or 
isozyme as an antigen and to look for a correlation of enzyme/isozjnne serum 
levels with tumor burden. 

Approach: Evaluate the immunodiagnostic significance of serum levels of UDP- 
galactosyltransf erase in human breast and ovarian carcinoma. 

Progress: Antisera directed against affinity-purified bovine UDP-galactosyl- 
transferase have been prepared in rabbits and partially immunologically charac- 
terized. These antisera have relatively high titered precipitating antibody 
to bovine galactosyltransf erase as assessed by the Ouchterlony double diffusion 
technique and inhibition of enzymatic activity. The antisera are highly cross- 
reactive with human UDP-galactosyltransf erase found in both normal serum and 
ascitic fluid obtained from a patient with ovarian cancer as determined by 
direct precipitation and inhibition of enzymatic activity. This cross-reactiv- 
ity has been exploited for the development of a sensitive and specific radio- 
immunoassay for human serum UDP-galactosyltransf erase with a detection level 
of 0.1-1.0 nanograms in the assay mixture. 

Work during the last 6 months has been focused on two areas: one, setting 
up and debugging of the NIH RIA data processing program which has been initi- 
ated to store and manipulate the data and two, performing a retrospective clin- 
ical study in which serum GT levels were quantitated by RIA on a limited number 
of patients with Stage IV breast carcinoma. These samples were initially col- 
lected and GT levels measured by kinetic assay. In this limited study, the GT 
levels were quantitated by the newly developed RIA and were in good agreement 
with the quantitation by kinetic assay. A more systematic and comprehensive 
study has been initiated to determine the degree of correlation of human serum 
GT levels with tumor burden, stage of disease, response to therapy, and pro- 
gression of tumor growth. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



218 



• 



CONTRACT RESEARCH SUMMARY 

Title: Clinical Evaluation of Immunodiagnostic Tests for Cancer 

Principal Investigator: Dr. John Fenimore Cooper 

Performing Organization: Kaiser Permanente Medical Group 

City and State: Los Angeles, CA 

Contract Number: NOl-CB-74169 

Starting Date: 9/16/77 Expiration Date: 7/15/81 

Goal: Evaluation of radioimmunodiagnostic tests for prostatic cancer. 

Approach: Accumulation and assessment of clinical oncologic data for prostatic 
cancer with a solid phase radioimmunoassay technique for prostatic acid phos- 
phatase (RIA-PAP); continuing development of the RIA-PAP technique as a poten- 
tial screening population of elderly males over the age of 50 years at risk for 
the disease. 

Progress: Analysis of the data obtained from the screening study of nearly 
6,000 men was continued with the following new findings. (1) In normal indi- 
viduals the mean RIA value obtained is independent of age. This implies that 
no age adjustment of the RIA value is required in clinical practice. (2) In 
individuals with benign prostatic hypertrophy (BPH) a similar invariance of the 
RIA value with age is observed. Thus, no adjustment of the observed RIA value 
is necessary in interpretation of data in BHP patients. (3) The number of 
cases detected per 1,000 cases screened increases dramatically over the age of 
65 years. These data should allow us to quantitatively limit any future 
screening to individuals in this age category. (4) A careful review of the 
clinical charts of individuals detected in the screening program demonstrates 
that some subjects had previously suspected or diagnosed history of prostatic 
disease. Most of these individuals demonstrated stage 4 disease at the time 
of the screening procedure. Exclusion of these individuals markedly shifts the 
stage distribution of detected disease to earlier stages (2 and 3). Therefore, 
the years of life saved by earlier detection of disease stages due to screening 
appears to be increased. Value of the screening procedure is greatly enhanced 
by these findings. Work currently in progress will provide detailed data 
description of individuals in the normal, BPH, and disease categories. Parame- 
ters of specificity and predictive value of a positive test will be examined. 
Finally, the value usually chosen to designate suspicion of disease (RIA value: 
6.5 mg/ml) will be varied between 7 and 10, and the impact of such changes on 
specificity and predictive value will be examined. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 

219 



CONTRACT RESEARCH SUMMARY 

Title: Diagnostic Applications of Antibodies to Melanoma 

Principal Investigator: Dr. Peter Hersey 

Performing Organization: Kanematsu Institute 

City and State: Sydney, Australia 

Contract Number: NOl-CB-74120 

Starting Date: 9/30/77 Expiration Date: 9/29/81 

Goal: Determine whether there is a correlation between the clinical course of 
melanoma growth and levels of serum antibodies. 

Approach: Employ lymphocyte-dependent cytotoxic antibody (LDA) assays. 

Progress: During the first 39 months of this study, 414 patients with local- 
ized melanoma were studied. Of these, 228 had LDA activity detected against 
allogeneic cultured melanoma cells. In 139 patients a rise in LDA activity was 
noted after surgical removal of the melanoma followed by a gradual delay to 
undetectable levels by 2-3 months. Twenty-seven patients had detectable anti- 
body to melanoma cells before removal of melanoma which remained unchanged 
after surgery. Twenty of the latter were multiparous women. Of the 248 pa- 
tients with no antibody detectable in their sera, 62 had LDA activity detecta- 
ble in the IgG fractions of sera obtained by gel filtration of acidified 
'unblocked' sera. Sera from 23 of 124 patients with non-melanoma carcinoma had 
LDA activity to allogeneic melanoma cells. All but one of the sera from this 
latter group of patients also reacted with allogeneic cells from cell lines 
established from breast and bladder. carcinoma and the Chang liver cell line and 
were not specific for melanoma. This contrasted with the results of studies 
on the melanoma patients in that 158 of the 228 positive sera did not react 
with cells from non-melanoma cultures. Studies on the specificities of these 
sera are in progress. Thirty-eight patients, studied at the time of initial 
presentation, have had recurrences. No significant differences in recurrence 
rate in relation to LDA status is yet apparent. Trends for a lower rate of 
recurrence in patients with high constant LDA are, however, apparent. Much 
longer follow-up periods are required before the importance of LDA in relation 
to recurrences from melanoma will be known. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: $33,900 



220 



t 



CONTRACT RESEARCH SUMMARY 

Title: Innovative Techniques for Passage of Colonoscope 

Principal Investigator: Dr. John A. Coller 

Performing Organization: Lahey Clinic Foundation 

City and State: Boston, MA 

Contract Number: NOl-CB-74212 

Starting Date: 9/26/77 Expiration Date: 9/25/81 

Goal: To develop a means for improving early detection of cancer on endoscop- 
ically approachable surfaces of the colon in order to improve life expectancy. 

Approach: A colonoscope advancing device will be developed to enhance the ease 
and speed of passage of currently available colonoscopes. Modeling studies and 
bench testing will be conducted during the initial year to determine which of 
several possible designs may be feasible. Further development of the colono- 
scope advancing device design will be carried out during the second year aided 
by physiological studies made to characterize as many physical parameters of 
the human colon as possible. Suitable devices will be given preliminary clin- 
ical tests during the latter part of the second year. After further optimiza- 
tion the clinical prototype propulsive devices will be tested by clinical 
trials in the third year. 

Progress: During the first year, efforts were directed towards defining and 
measuring physiologic characteristics of colon function that relate to the 
intubation problem. From these observations, together with a mathematical 
modeling approach, various bench models were developed for testing discrete 
aspects of the problem. The various techniques in the construction of fibro- 
elastomeric structures were developed. During the second year, fibroelasto- 
meric structures of various configurations were constructed and submitted to 
both bench and animal testing. A major problem which became apparent at this 
point was the disparity between the various bench models and the more compliant 
animal colon. Consequently, the major effort during year three has been to use 
the animal preparation as the main evaluatory system with f ibroelastomeric 
devices that are designed to accept rather than combat the compliant nature of 
the colon. At present, suitable f ibroelastomeric constructions are being pre- 
pared for preliminary evaluation in humans. 

The structural configuration of the surface envelope has been altered in 
order to minimize the longitudinal as well as torsional resistance. The axial 
driving concept has been replaced by a torsional mechanism directed at a random 
lumen-seeking tip. Further experimental study as well as trial human applica- 
tion is anticipated during year four. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



221 



CONTRACT RESEARCH SUMMARY 
Title: Detection and Localization of Early Lung Cancer 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-53886 
9/27/74 



Dr. Robert S. Fontana 
Mayo Foundation 
Rochester, MN 



Expiration Date: 9/26/81 



Goal: To test new methods of diagnosis of early lung cancer and to assess sur- 
vival of patients with lung cancer detected by these methods. 

Approach: A study population of 9,211 subjects was obtained following initial 
screening of 11,001 non-volunteers. Candidates were Mayo Clinic outpatients 
who were men, 45 years old or more, smoking one pack of cigarettes or more 
daily. None of those accepted into the study had a history or suspicion of 
respiratory cancer on entry into the Clinic; all had a life expectancy of at 
least 5 years, and all were judged capable of tolerating pulmonary resection 
(at least lobectomy). Initial screening consisted of chest radiographs, 3-day 
sputum cytology tests and lung health questionnaires. This screening yielded 
91 unsuspected ("prevalence") lung cancers, 17 detected by cytology alone, 59 
by radiography alone, and 15 by both tests. Radiographically "occult" cancers 
were localized f iberbronchoscopically. The 9,211 study subjects who had "nega- 
tive" initial screens were randomized into a close-surveillance group, for whom 
rescreening every 4 months was urged , and a standard surveillance ("control") 
group advised, but not reminded , to undergo rescreening once a year. A popula- 
tion of post-surgical AJC stage I lung cancer patients is also being evaluated, 
as are hematoporphyrin derivative (HpD) and cryotherapy as techniques for 
"mapping" and treating carcinoma-in-situ. 

Progress: As of April 1, 1981, the 4-monthly surveillance group and the "con- 
trol" group had been observed more than 27,000 man-years, and 302 new ("inci- 
dence") cases of cancer of the respiratory tract had been detected, 63 involv- 
ing the upper airway and 239 the lungs. In the 4-monthly surveillance group 
there were 135 new cancers (14 of which were detected by cytology only) and 68 
lung cancer deaths. Survival data in this group are encouraging. In the con- 
trol group there were 104 new lung cancers and 68 lung cancer deaths, with a 
tendency toward more traditional lung cancer survivorship. Thus, once lung 
cancer has been detected, survival in the 4-monthly surveillance group is con- 
siderably better than among the controls. So far there has been no decrease 
in total deaths from lung cancer in the close-surveillance group compared to 
the control group. Prolonged follow-up is essential before final conclusions 
and specific recommendations can be made. As "incidence" cases continue to 
accrue, more detailed analyses of data are feasible. Attention is currently 
directed toward evaluating the results of screening by cell type of tvimor and 
by modality of detection. The post-surgical AJC stage I lung cancer study in- 
dicates that about 70% of those with non-small cell cancer survive 5 years. 
The HpD study has detected several squamous cancers that were both radiograph- 
ically and endoscopically occult. Cryotherapy continues to be used in selected 
cases. 

Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $650,000 



222 



• 



CONTRACT RESEARCH SUMMARY 

Title: Carcinoembryonic Antigen in the Diagnosis of Bowel Cancer 

Principal Investigator: Dr. Vay Liang W. Go 

Performing Organization: Mayo Foundation 

City and State: Rochester, MN 

Contract Number: NOl-CB-23854 

Starting Date: 6/15/72 Expiration Date: 6/14/82 

Goal: To determine the usefulness of CEA in the diagnosis of bowel cancer. 

Approach: (a) Measure CEA levels in blood and other specimens stored at NCI- 
Mayo Clinic Human Serum Bank (CB-84258), Mayo Lung Cancer Project (NCI-CB-22000) 
and other contracts with the Diagnosis Program of the Division of Cancer Biology 
and Diagnosis; (b) continue to collect and provide colonic malignant tissue for 
different investigators in projects dealing with carcinoembryonic antigens; (c) 
complete the work done over the last five years in the continuation of the eval- 
uation of CEA as a biologic marker in patients undergoing chemotherapy, radio- 
therapy, and immunotherapy of advanced gastrointestinal cancer. 

Progress: (a) Progress related to the contract's service function activity . 
Serum or plasma levels have been determined in 7,984 specimens from NCI Serum 
Plasma Bank including those sent in from other investigators. These samples 
were run at 20% of the commercial cost. Comparative studies with other method- 
ologies for measuring CEA including the enzyme immunoassay were carried out. 
The comparison of values of CEA to other tumor-associated antigens was done with 
investigators from other institutions and has resulted in several publications. 
Statistical analyses of the data over the last three years were included in the 
Workshop on Immunodiagnosis of Human Cancer, Statistical Analyses, Part II. 
Results and statistical analyses are being performed on coded serum panels which 
were sent last year to 17 investigators with potentially useful diagnostic tests 
for cancer. 

(b) Progress report on the evaluation of CEA as a biological marker . Data 
are still being collected for the evaluation of CEA as a biologic marker in pa- 
tients following resection of gastrointestinal cancer with curative intent, in 
patients undergoing chemotherapy, radiotherapy and immunotherapy for advanced 
gastrointestinal cancer and in lung cancer patients. Preliminary data have been 
analyzed for publication purposes. 



Project Officer: Mr. Louis P. Greenberg 
Program: Diagnosis 
FY 81 Funds: $49,500 

223 



CONTRACT RESEARCH SUMMARY 
Title: Maintenance of the NCI Serum Diagnostic Bank 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-84258 
9/30/78 



Dr. V. L. W. Go 
Mayo Foundation 
Rochester, MN 



Expiration Date: 6/30/81 



Goal: To establish and maintain a bank, of sera from patients with cancer, with 
benign diseases and from normal individuals, for evaluating immunodiagnostic 
tests of potential clinical usefulness in the diagnosis of cancer. 

Approach: Make necessary serum samples available for evaluation of immunodiag- 
nostic tests for cancer. Serve as a central facility for storage of serum and 
plasma specimens collected by other contractors in the Tumor Immunology- Immuno- 
diagnosis Program. 

Progress: In 1981 through April 1, 5,342 vials of sera were collected by the 
central facility for a total of 147,637. In 1981, 1,437 vials were shipped out 
to investigators for a total of 63,187. Presently there are 84,450 vials from 
the central facility and 160,701 vials from various screening projects for a 
total of 245,151 vials stored at the Mayo-NCI Serum Diagnostic Bank. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: $85,810 



224 



CONTRA.CT RESEARCH SUMMARY 

Title: Reconstruction Algorithms for Dose Reduction in X-Ray Computed 

Tomography- 
Principal Investigator: Dr. Richard A. Robb 
Performing Organization: ^ Mayo Foundation 
City and State: Rochester, MN 

Contract Number: NOl-CB-84235 

Starting Date: 9/29/78 Expiration Date: 9/28/81 

Goal: To develop and evaluate new and/or improved reconstruction methods 
directed toward reducing the x-ray dosage required to obtain diagnostically 
accurate information from CT scanning procedures used in cancer diagnosis. 

Approach: A variety of approaches will be investigated for their contributions 
to improved image reconstruction with reduction in x-ray dosage. These will 
include studies for tailoring and optimizing convolution, least square, and 
iterative algorithms for low x-ray dose data; use of a priori information; 
noise reduction techniques; minimizing required field of view; non-linear spa- 
tial smoothing; and optimizing algorithms based on penalty functions and multi- 
plier methods for constrained optimization. These methods will be tested on 
realistic mathematical simulations and on actual patient CT scan data. The 
results will be quantitatively evaluated using standard image quality measure- 
ment criteria and by comprehensive psychophysical (ROC) studies. 

Progress: Different types and versions of reconstruction methods have been 
compared. Preliminary results in the use of positivity constraints indicate 
improvements in accuracy and speed of convergence for iterative type recon- 
structions but questionable effects for convolution methods. The maximum 
entropy algorithm has been applied to simulated low dose CT data containing 
complex lesions in uniform fields. Evaluation of the reconstruction has not 
confirmed the notion that such an algorithm is more effective than others in 
reducing the number of "false positives." Preliminary studies with algorithms 
for limited field of view indicate that relatively low contrast lesions are 
detectable and measurable in the field of view at low x-ray doses. Preliminary 
results with selective non-linear smoothing techniques indicate significant 
improvement in image signal-to-noise with potential for improvement in diagnos- 
tic quality. A survey of the field of optimizing algorithms has been prepared. 
Extensions of previously known methods for quadratic optimization have been 
produced. A preliminary ROC study has been performed on images of a realistic 
head phantom, with superimposed low-contrast "lesions," reconstructed using the 
standard convolution method on projection data generated with photon counts 
both typical of and considerably less than present day CT x-ray scanners. Two 
other algorithms have been selected for evaluation using these data — weighted 
least square and an iterative optimizing algorithm which combines notions of 
regularization, penalty functions, Bayesian and multiplier methods. These 
algorithms will also be applied to actual clinical CT data of patients with 
lesions. 

Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



225 



CONTRACT RESEARCH SUMMARY 

Title: Collection of Serial Serum Samples from Cancer Patients 

Principal Investigator: Dr. Carl Feit 

Performing Organization: Memorial Hospital for Cancer and 

Allied Diseases 
City and State: New York, NY 

Contract Number: NOl-CB-04350 

Starting Date: 9/30/77 Expiration Date: 9/29/82 

Goal: Collect serial serum specimens from cancer patients and benign disease 
controls for evaluation of potential immunodiagnostic tests for cancer. 

Approach: Collect serial specimens from patients with melanoma, ovarian car- 
cinoma, lung carcinoma, and Hodgkin's disease plus suitably matched patients 
with benign skin lesions, benign adnexal masses, a history of heavy smoking, 
and benign lymphadenopathy to serve as controls. 

Progress: This contract is completing the fourth year of a program for collec- 
tion of serial serum specimens from cancer and control patients for use in 
evaluating potential immunodiagnostic tests for cancer. A total of 277 pa- 
tients have been entered, including 155 patients and 122 controls. By diagno- 
sis, the respective figures for patient/control entry are: ovarian, 37/33; 
lung, 32/20; Hodgkin's disease, 55/34; and melanoma, 31/25. An average of 44 
blood samples are collected monthly. The total number of samples in the col- 
lection to date is 1,537. The patient population projected for completion of 
study is 325. To achieve this population it is anticipated 110 patients will 
have to be recruited. Progress has been made in data entry and computer con- 
trol of data. Consultations are under way relating to the epidemiologic sound- 
ness of the collection and biostatistical approaches for handling the data 
generated by testing are being developed. All serum samples have been tested 
this year for a single marker, lipid-bound sialic acid (LSA) . These data are 
being used by the biostatisticians to probe the utility of the collection for 
evaluating new tumor markers. 



I 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: $135,104 



226 



CONTRACT RESEARCH SUMMARY 

Title: Lung Cancer - Early Detection, Localization and Therapy 

Principal Investigator: Dr. Myron R. Melamed 

Performing Organization: Memorial Hospital for Cancer and 

Allied Diseases 
City and State: New York, NY 

Contract Number: NOl-CN-45007 

Starting Date: 9/01/73 Expiration Date: 7/31/81 

Goal: To evaluate sputum cytopathology as a supplement to annual chest x-rays 
in detecting pulmonary neoplasms at an "early stage" and to evaluate the effi- 
cacy of techniques for prompt localization of radiologically occult lung cancer 
(e.g., before progression to x-ray positive); in general, to evaluate the effi- 
cacy of such screening to reduce lung cancer mortality. 

Approach: Over a 3-year period, 5,000 test subjects and 5,000 control subjects 
have been entered into this study. Each subject will receive active screening 
for at least 5 years, all followed for at least an additional 5-year period. 
This will be conducted according to a protocol developed in conjunction with 
the Johns Hopkins University and the Mayo Foundation. 

Progress: Recruitment for the Memorial Sloan-Kettering Cancer Center National 
Lung Program was completed in January 1978. Total enrollment is 10,040. Par- 
ticipants were randomly assigned to study group (A) (4,969), receiving annual 
chest x-rays and 4-monthly sputum; and to control group (B) (5,071) receiving 
annual x-rays only. There have been a total of 217 confirmed lung cancers 
identified so far, 105 in group A and 112 in group B. The principal mode of 
detection in the A group was by cytology in 19 cases, x-ray in 49 cases and 
both techniques in 12 cases. Of the 19 cases detected by cytology, 13 were 
Stage or I; of the 61 cases detected by radiology (or radiology and cytology) 
in group A and the 72 cases detected by radiology in group B, a total of 133 
cases; there were 65 in Stage I. There were 65 interval cancers diagnosed fol- 
lowing symptoms or signs or by x-rays taken outside of the routine screening 
program; only 11 of these were Stage I, and 21 were oat cell cancers. 

Among the 105 lung cancers appearing in group A, 30 were prevalence cancers 
for a prevalence rate of 6.0/1,000 and 75 were incidence cancers for an inci- 
dence rate of 3.0/1,000/year. In group B the 112 lung cancers included 23 pre- 
valence for a prevalence rate of 4.6/1,000 and 89 incidence cases, for an inci- 
dence rate of 3 .5/1,000/year. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $900,000 



227 



CONTRACT RESEARCH SUMMARY 
Title: Evaluation of Screening Methods for Endometrial Cancers 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-84233 
9/30/78 



Dr. Leopold G. Koss 
Montefiore Hospital 
Bronx, NY 



Expiration Date: 9/29/81 



Goal: To carry out an extensive study to test the feasibility of screening for 
endometrial cancers in asjnnptomatic women by means of uterine sampling. 

Approach: Three thousand two hundred asymptomatic women over age 45 and up to 
age 70 years will be enrolled into the study. Sixteen hundred of the women 
will be in the age range 46 to 59 years and 1,600 will be in the age 60 and 
above. After history has been obtained and the examining procedure explained, 
each consenting examinee will receive a complete gynecologic examination, a 
vaginal pool smear, a cervical scrape smear, and an endocervical aspiration 
smear. Subsequently, the examinees will receive a Mimark or Isaacs (Curity) 
endometrial sampling randomly assigned by computer. The patient's acceptance 
of the procedure will be evaluated and recorded by the gynecologist upon com- 
pletion of the examination. Results of the laboratory examination will be com- 
municated by the contractor to the patient's primary physician and to the 
patient. 

Progress: The enrollment of the examinees began on January 2, 1979. From that 
date until June 30, 1980, 1,784 primary eligible examinees received the clini- 
cal and cytologic examinations and the endometrial sampling described. There 
were 100 additional women who either refused to sign the informed consent or 
who were otherwise not eligible. Approximately 84% of the examinees were white 
and approximately 82% were 50 years of age or older. The endometrial procedure 
was successfully implemented in about 92% of the women with virtually no com- 
plications. Of the 1,784 examinees 146 were recalled at least once to clarify 
atypical or abnormal findings. Amongst the group of 1,784 examinees there were 
8 endometrial carcinomas and 21 other endometrial abnormalities documented by 
histology (12 hyperplasias, 9 polyps). We also have failed in the identifica- 
tion of 1 confirmed endometrial cancer. As of January 1, 1980, a recall pro- 
gram of examinees seen 1 year before was begun. Between January 1 and June 30, 
259 women returned for a repeat examination, giving a return rate of 37%. Some 
of these returnees may require additional follow-up. The total number of pa- 
tients' visits during the reporting period was 2,189. It is of interest that 
5 breast cancers, 1 ovarian cancer, 1 carcinoma in situ of the vulva, and 12 
cervical Intraepithelial neoplasias were also diagnosed in this asymptomatic 
population. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $193,440 



228 



CONTRACT RESEARCH SUMMARY 



Title: Development of Electrophoretic Display Cell 



Principal Investigator: 
Performing Organization; 
City and State: 



Contract Number; 
Starting Date: 



NOl-CB-04341 
9/30/80 



Dr. Howard Sorkin 

North American Phillips Corp. 

Briarcliff Manor, NY 



Expiration Date: 3/29/82 



Goal: To produce a large area electrophoretic display system for X-ray imaging 
which is suitable for use in clinical radiological diagnosis including tumor 
imaging • 

Approach: The contractor will complete the production of an X-ray sensitive elec- 
trophoretic display system which was partially developed prior to the contract. 
The unit is basically a thin cell with a suspension of pigment particle in a dyed 
organic liquid held between electrode plates, one of which is transparent. Images 
produced by X-rays impinging on one of the electrodes, analogous to film sensiti- 
zation by X-rays, will be visualized in the display cell by appropriate selective 
deposition of the pigment on the electrode surface. The system has a memory which 
is erasable by polarization changes in the electrodes. The specifications of the 
display cell proposed by the contractor are as follows: limiting resolution, 8 
Ip/mm; photopic contrast, 10:1; monochromatic contrast, 25:1; linear shades of 
grey, > 500; saturation exposure, 0.5 mR; exposure for mid-grey level, 200 ^R; 
image formation time, 100 ms; image retention time, >24 hr; image size, 10" x 10". 

Progress: The development of improved PbO-binder layers has been separated into 
two distinct areas: mechanical methods of uniform layer deposition, and elucida- 
tion and optimization of the chemistry of the system. PbO-binder layers have been 
prepared by: rapid settling from dilute binder solutions; settling from viscous 
mixtures; direct application to a substrate using a blade to obtain correct layer 
thickness. The latter two methods give the most uniform layers as evidenced by 
radiographs of the dried, cured layers. To date, the most X-ray sensitive layers 
have been fabricated using alkydresin binders. These resins contain free organic 
acids which react with PbO to form lead salts. Efforts to determine whether these 
salts remain on the crystal surfaces and their effect on the X-ray performance of 
the binder layer are in progress. Two methods were developed for electrically 
evaluating PbO binder layers, viz., a dynamic (rotating) electrometer technique 
and a static capacitance technique. These techniques are being used to study the 
layers independently of the EPID cell. A method of permanently chemically bonding 
the stabilizer/charging agent to the pigment particles has been developed. Stable 
suspensions having the desired low pigment particle charge have been prepared 
using pigment modified by this method. Highly encouraging performance of X-ray 
imaging cells using these suspensions has been obtained. A method of removing 
impurities in commercially available dyes, by treatment with molecular sieves, has 
been developed. The purified dyes give suspensions with substantially lower con- 
ductivity and no electrohydrodynamic instability. Medical-quality X-ray equipment 
has been installed and approved for use by the contractor's radiation safety 
officer. 

Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



229 



CONTRACT RESEARCH SUMMARY 
Title: Innovative Techniques for Passage of Colonoscope 



Principal Investigator: 
Performing Organization: 
City and State: 



Contract Number: 
Starting Date: 



NOl-CB-74114 
9/26/77 



Dr. Max Epstein 
Northwestern University 
Evanston, IL 



Expiration Date: 9/25/81 



Goal: To develop safe and simple techniques and devices for early detection 
of small cancer lesions by means of endoscopic examination of the entire colon 
in order to improve life expectancy of postoperative patients and prevent or 
treat colonic cancer by endoscopic polypectomy. 

Approach: Several devices will be investigated to enhance safety and ease of 
advancement of currently available colonoscope to the cecum. Prototype design, 
fabrication, and preliminary tests in simulated environment models during the 
first year will be followed by animal tests of promising designs during the 
second year. Also during the second year, the most promising devices will be 
fabricated in an appropriate form for clinical testing. During the third year 
the main efforts will be directed at clinical trials of one or more of the 
developed devices. 

Progress: Three devices and several techniques have been developed. 1. The 
endoscopic extender, a miniature endoscope which can be introduced through the 
ancillary channel of the conventional endoscope, has been shown to be useful 
in animal tests and in patients. The results were published and further work 
awaits the design of a somewhat smaller device which can be more conveniently 
passed through most current colonoscopes. 2. The inflatable liner has been 
shown to be effective in tests in dogs and also in patients. Most recently, 
the device was modified to allow for the liner to be retracted, thereby facil- 
itating its insertion and passage through bends and obstructions in the colon. 
A prototype liner utilizing water instead of air is currently being tested. 
3. Several versions of an overtube were constructed which should greatly facil- 
itate safe passage of the colonoscope, in particular, in the case of repeated 
insertion and withdrawal such as employed in multiple polypectomies. 4. A num- 
ber of techniques to safely pass the presently available colonoscopes have been 
developed and will be described in a forthcoming report and will be submitted 
for publication. 

In order to allow for the appropriate testing in patients, the research 
project has been extended for an additional (fourth) year at no cost. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



230 



CONTRACT RESEARCH SUMMARY 
Title: Sera Collection from High Cancer Risk Populations 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Henry Altschuler 
Philadelphia Geriatric Center 
Philadelphia, PA 



Contract Number: 
Starting Date: 



NOl-CB-33914 
6/25/73 



Expiration Date: 11/24/80 



Goal: To collect serial serum specimens from populations at high risk of con- 
tracting malignant disease for the purpose of developing and evaluating immuno- 
diagnostic screening tests for human cancer. 

Approach: Demographically stable populations at high risk of developing human 
malignancies in the colon, lung, breast, prostate, bladder, pancreas, etc. will 
be followed on an annual basis. This includes clinical evaluation and serum 
collection. The sera will be banked and kept on the patients so that informa- 
tion obtained from the stored sera can be correlated with development of any 
particular cancer. 

Progress: Over the past 6 years and 10 months, a total of 4,211 serum speci- 
mens (divided into ten 1-ml aliquots) have been collected, processed, logged 
with assigned numbers on a chronological basis, frozen, and shipped frozen in 
dry ice to the Regional Serum Storage Center at the Mayo Clinic in Rochester, 
Minnesota. In addition, there is a patient card index, alphabetically orga- 
nized by patient's name, and including age, sex, hospital MDI number, address, 
diagnosis, serum numbers (a label for each date of collection of blood placed 
on card), and date of collection of blood. There are additional cards which 
contain such information as the date of the last physical examination and in- 
formation concerning such conditions as asthma, allergy, rheumatoid arthritis, 
desensitization therapy, immunization (influenza vaccine), and blood transfu- 
sions. Also maintained are a cancer index and a death index card file with 
significant findings at autopsy if performed. Data from these files have been 
collated on special computer adaptable forms supplied by NCI. An overall 
detailed clinical summary has been completed. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



231 



CONTRACT RESEARCH SUMMARY 

Title: Tumor-Specific Antigens in Diagnosis and Management of Cancer Patients 

Principal Investigator: Dr. T. M. Chu 

Performing Organization: Roswell Park Memorial Institute 

City and State: Buffalo, NY 

Contract Number: NOl-CB-33858 

Starting Date: 6/25/73 Expiration Date: 12/24/80 

Goal: To study the relationship of CEA in the evaluation of recurrence and 
survival for cancer patients with curative surgery. 

Approach: Examine the prognostic value of preoperative and follow-up CEA in 
relation to other preoperative variables; study the distribution profiles of 
CEA at surgery, during follow-up period, and at recurrence; implement a plan 
for patients whose risk of recurrence is greatest based upon preoperative and 
serial CEA; develop predictive models which integrate the CEA levels with other 
prognostic variables to determine the probability of recurrence in order to 
recommend adjuvant therapy. 

Progress: An analysis of long-term follow-up CEA data and baseline data is 
under way on 74 patients with colorectal cancer and on 130 patients with lung 
cancer who underwent resections for cure. Of 74 colorectal cancer patients, 
35 had recurrence of disease and 32 died. Preoperative CEA was found to be 
correlated with Dukes' classification, and had significant prognostic value 
between 9 and 18 months post surgery. Thereafter, Dukes' classification was a M j 
more significant predictor of disease recurrence. Matched-pair techniques 
revealed that CEA was often elevated in anticipation of recurrence, sometimes 
as early as one year prior to recurrence. The accuracy for using follow-up CEA 
events in predicting recurrence was restricted due to high false positive rates 
and low true positive rate. Further analysis is under way. Among patients 
experiencing recurrence, although forewarned by CEA elevations, no significant 
correlation was found between the time interval and risk of recurrence. The 
correlation of CEA evaluations alone with the time of recurrence was not strong 
enough to suggest a second-look surgery without other supporting evidence. In 
collaboration with NCI, a time-dependent Cox statistical model is being devel- 
oped to re-investigate the relationship between follow-up CEA and disease 
recurrence. In patients with resectable lung carcinoma, CEA has more precise 
value in distinguishing, at an early date (sometimes at surgery), cured pa- 
tients from those who would fail subsequent to surgical resection because of 
recurrent disease. Adjuvant therapies could thus be recommended for those pa- 
tients with high risk for recurrence. In summary: (i) preoperative CEA is 
positively correlated with Dukes' classification; (ii) preoperative CEA adds 
significant information on Dukes' classification in the estimation of recur- 
rence rates; (ili) postoperative CEA assays taken later in the clinical evalua- 
tion process carry the most prognostic information; (iv) CEA often rises in 
anticipation of recurrence; and (v) follow-up CEA events depending on multiple 
CEA observations have some accuracy in predicting recurrences but are of 
limited value in predicting the exact times of recurrence. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 

232 



^ 



CONTRACT RESEARCH SUMMARY 

Title: Application of Digital Image Processing Techniques to Cytology Automa- 
tion 



Principal Investigator: 
Performing Organization: 



City and State: 

Contract Number: 
Starting Date: 



NOl-CB-74190 
9/30/77 



Dr. James W. Bacus 
Rush-Presbyterian-St . Luke's Medical 

Center 
Chicago, IL 



Expiration Date: 9/29/81 



Goal: Development of optimal algorithms for cell classification applicable to 
an automated digital image processing system. 

Approach: The contractor shall conduct a comprehensive study to determine 
single cell vs. specimen classification accuracies for image processing algo- 
rithms and for cytotechnologists. This will determine how well the machine 
algorithms are performing. Five experimental tasks will be done: 1) sample 
acquisition, 2) cell acquisition, 3) observer recognition, 4) cell classifica- 
tion algorithms development, and 5) analysis and evaluation of results. 

Progress: Patient selection and sample acquisition are essentially completed. 
Fabrication of the cell acquisition system is finished. Cells are being 
acquired routinely with this equipment. Programming of cell recognition algo- 
rithms continues with current emphasis on texture analysis and single cell 
classification. The observer recognition experiment is under way for single 
isolated cells out of context with background information and for single cells 
using the locally surrounding background information. Although the studies 
have not been completed — to date 23 observers have classified the same 1,650 
cells, both with and without background information (from a targeted 10,000 
cells, over 50 cases) — the preliminary results clearly indicate low classifica- 
tion accuracies for observers when they rely only on the information they can 
extract from single cells one-at-a-time. Since significantly abnormal (or 
positive) cells have a very low occurrence, these poor single cell performance 
characteristics of observers strongly suggest that traditional cervical cytol- 
ogy screening does not rely heavily on systematic location and classification 
of single cells one-at-a-time. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



233 



CONTRACT RESEARCH SUMMARY 
Title: Isolation and Tissue Culture of Human Tumor Cells 



Principal Investigator: 
Performing Organization: 

City and State: 

Contract Number: NOl-CB-43854 
Starting Date: 8/08/73 



Dr. J0rgen E. Fogh 

Sloan-Kettering Institute for Cancer 

Research 
Rye, NY 



Expiration Date: 8/07/81 



Goal: To develop further procedures for encouraging tumor cell growth in cul- 
ture and to perform tests necessary to define standard cell lines from various 
tumors. 

Approach: Human tumor cell lines are established in culture from surgical 
specimens and are collected from other investigators. They are characterized 
in this laboratory as to malignancy, cell type and special features as needed 
for individual investigations. 

Progress: The contract supports a resource in the form of an extensive bank 
of human tumor lines grown in vitro . Cell lines are preserved after as few 
passages as possible, and are characterized in terms of morphology, nutritional 
requirements, growth in vitro , chromosomal characteristics and poliovirus sus- 
ceptibility. They are monitored for microbial contamination. Analyses of 
polymorphic enzyme patterns enable the investigators to determine possible 
cross-contamination with other cell lines and to exclude such contamination 
when unique phenotype combinations are observed. The 426 cultured human tumor 
cell lines are available to other investigators. The bank now includes 69 
melanomas, 31 carcinomas of the breast, 20 brain and nervous tissue tumors, 19 
carcinomas of the bladder, 18 of the colon, 17 of the lung, and 10 of the kid- 
ney. Five hundred and sixty-four lines have been made available to other 
investigators in a 12-month period (1980), plus 238 for the period of 1/1/81 
through 5/4/81, indicating a good utilization of the resource. 



€i 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



234 



t 



CONTRACT RESEARCH SUMMARY 

Title: Ultrasonic Probes Inserted Through Endoscopes in Cancer Diagnosis 

Principal Investigator: Mr. James L. Buxton 

Performing Organization: SRI International 

City and State: Menlo Park, CA 

Contract Number: NOl-CB-74136 

Starting Date: 9/30/77 Expiration Date: 1/31/81 

Goal: To develop and clinically test an ultrasonic imaging system which will 
provide extremely high resolution images of the anatomy in the thorax and upper 
abdomen. 

Approach: Currently available imaging systems have inadequate resolution to 
diagnose many diseases in organs in the middle of the torso. For instance, 
conventional imaging systems cannot diagnose pancreatic cancer until very 
advanced stages. The thrust of this project is to develop an ultrasonic imag- 
ing system with its transducer on the tip of an endoscope, so that internal 
organs can be imaged through the wall of the gastrointestinal tract. The 
resulting proximity to internal organs permits high resolution images and, in 
addition, eliminates the shadowing by bowel gas which plagues conventional 
ultrasonic systems. 

Progress: By the end of the first year, the basic ultrasonic imaging system 
had been developed and demonstrated. The system is organized around a 64 ele- 
ment, 10 MHz, linear array which is 30 mm long. The 3 x 4 cm image is produced 
in a B-scan format at approximately 30 frames per second. Dynamic focusing on 
both transmit and receive provides submillimeter resolution in both dimensions 
throughout the field of view. Midway through the second year of the project, 
a prototype endoscopic imaging system was installed at the Mayo Clinic for 
tests with dogs. This first system included an endoscope with an outside diam- 
eter of 13 mm, side-looking optics and ultrasound, and a rigid tip length of 
80 mm. This system demonstrated that intragastric imaging provides very high 
resolution imaging of the anatomy near the upper GI tract. Among the organs 
imaged were: heart, liver, both kidneys, gallbladder and biliary tree, spleen, 
and abdominal vasculature. Early in the third year of the project, an improved 
endoscope was completed. Changing the optics to look forward allowed the rigid 
tip length to be reduced to 40 mm. With this unit, preclinical trials \^ere 
started in November 1979. In human control subjects, this system has imaged 
all of the organs that the first system imaged in dogs and, in addition, has 
demonstrated the entire pancreas. The system has successfully been used to 
diagnose a variety of diseases, including chronic pancreatitis, pancreatic 
abscess, pancreatic cancer, liver cancer, and stomach ulcers. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $50,000 



235 



CONTRACT RESEARCH SUMMARY 

Title: Gelatin Encapsulated Microbubbles as Ultrasonic Contrast Agents 

Principal Investigator: Dr. Barbara A. Carroll 

Performing Organization: Leland Stanford Jr. University 

City and State: Stanford, CA 

Contract Number: NOl-CB-14337 

Starting Date: 5/15/81 Expiration Date: 5/14/83 

Goal: To determine the effectiveness of gas-filled microbubbles as contrast 
agents to be used in diagnostic ultrasound. 

Approach: Appropriate sized gas-filled microbubbles will be produced and in 
vitro testing systems will be conducted. In vivo testing will be done in 
tijmor-bearing animals to determine the- ultrasound contrast characteristics of 
the microbubbles. Acute and chronic toxicity studies on promising microbubbles 
will also be done. 

Progress: New Project. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: ^212, 384 



236 



CONTRACT RESEARCH SUMMARY 
Title: Alterations in Cellular and Humoral Anti-tumor Immune Reactivities 



Principal Investigator: 
Performing Organization: 
City and State: 



Dr. Isaac P. Witz 
Tel Aviv University 
Tel Aviv, Israel 



Contract Number: 
Starting Date: 



NOl-CB-74134 
9/30/77 



Expiration Date: 7/29/81 



Goal: Study alterations in the natural cellular and humoral reactivity pat- 
terns in mice at high risk, of developing carcinogen-induced or endogenous 
virus-induced malignancies as well as serological reactivity of such mice 
toward antigens associated with the relevant tumors. 

Approach: Characterize early alterations in naturally-occurring cellular and 
humoral anti-tumor immune reactivities in the following animal model systems: 
spontaneous tumors in C3HeB mice; mammary tumors induced by DMBA in BALB/c 
mice; and lung adenomas in urethan-treated BALB/c mice. 

Progress: This contract is in its fourth year of work, studying alterations 
of serological and cellular reactivity patterns in mice at high risk of devel- 
oping malignancies. The serologic pattern of normal mice has been established 
to serve as a basis for comparison of altered reactivity as carcinogen and 
viral-induced malignancies develop. The sequential serologic analysis of 
urethan-treated mice (those which developed lung adenomas and those which did 
not) throughout the carcinogenesis latency and the tumor-bearer periods has 
been completed. Sera from serial blood samples were analyzed by complement- 
dependent lysis (CdL) assays using 4 different tumor target cells. A statisti- 
cal analysis has shown that the CdL of one of these targets mediated by serum 
of urethan-treated mice that developed tumors differed significantly from that 
of mice which did not develop tumors. This difference was first noted during 
the carcinogenesis latency period. 

A time course study of NK activity in DMBA-fed mice was recently completed. 
Very early following DMBA administration the number of cells in the spleen of 
the treated animals was decreased to about 50% of normal values. The NK activ- 
ity (calculated on a per cell basis) was also significantly decreased. These 
two effects caused a 4-6-fold decrease in the number of lytic units in the 
spleen of DMBA-treated animals as compared to normal controls. The number of 
splenocytes was spontaneously restored to normal values about 3 months follow- 
ing DMBA administration and before most animals developed tumors. On the other 
hand, the NK activity remained depressed throughout the entire carcinogenesis 
latency period. A further significant decrease in NK activity occurred in 
those animals which developed primary tumors. This further depression seemed 
to be secondary to tumor development and was mediated by suppressor cells. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



237 



CONTRACT RESEARCH SUMMARY 

Title: Radioisotope Surface Markers and Detectors for Endoscopic Techniques 

Principal Investigator: Dr. James M. Woolfenden 

Performing Organization: University of Arizona Medical School 

City and State: Tucson, AZ 

Contract Number: NOl-CB-64012 

Starting Date: 6/30/76 Expiration Date: 12/29/80 

Goal: To establish a means for improving early detection of small cancer 
lesions on endoscopically approachable surfaces in order to improve the life 
expectancy of the patient. 

Approach: A system will be developed for endoscopic detection of small cancer 
lesions in mucosal surfaces using tumor-seeking radioisotopic markers. A 
miniature radiation detector system suitably matched to emission characteris- 
tics of the isotopic markers will be developed. During the first year, work 
will center on development and testing of tumor-seeking markers. Initial 
detector design and construction will begin. During the second year, develop- 
ment of markers will continue, and candidate detector systems will be tested 
using a ttimor phantom system. The period from the beginning of the third year 
to the termination of the contract will concentrate on patient studies to 
assess the sensitivity, specificity and accuracy of the tumor markers and 
detector system in localizing early cancer lesions. 

Progress: Scintillation detectors made of thallium-activated sodium iodide and 
thallium-activated cesium iodide and a cadmium telluride semiconductor detector 
have been constructed. All have performed satisfactorily in tumor phantom 
studies, with better performance by the Nal(Tl) and CdTe detectors than the 
CsI(Tl). The NAI(Tl) detector is being used in clinical patient studies. The 
tumor-seeking marker is cobalt-57 bleomycin. External imaging is performed 24 
hours after intravenous injection of 1 mCi Co-57 bleomycin; standard fiberoptic 
bronchoscopy is then done. The detector, coupled to an external photomulti- 
plier tube by a flexible fiberoptic light guide, is passed through the broncho- 
scope biopsy channel, and counts are taken at sites throughout the bronchial 
tree. Data for 36 patient studies for which clinical follow-up information is 
available indicate detector sensitivity of 82%, specificity of 79%, and accu- 
racy of 82% in locating tumors. Sensitivity of bronchoscopy including cytology 
and the detector combined was 91%; the two techniques appear to be complemen- 
tary. Bronchoscopically invisible submucosal tumors and occult sources of 
malignant cytology can be found using the detector system in some patients. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



238 



CONTRACT RESEARCH SUMMARY 

Title: New Stains and Other Optical Markers for Cytopathologic Specimens in 
Suspension 

Principal Investigator: Dr. Brian H. Mayall 

Performing Organization: University of California 

City and State: Livermore, CA 

Contract Number: Y01-CB-40300 

Starting Date: 6/01/74 Expiration Date: 7/31/81 

Goal: To develop and evaluate stains that can be used in flow cytometry to 
differentiate noirmal from dysplastic and malignant gynecologic specimens. 

Approach: The contractor shall: 1. Utilize flow cytometry to analyze 400 
gynecologic samples stained in suspension with chromomycin A3. 2. Employ com- 
puter analysis to evaluate cellular DNA content and orthogonal light scatter 
signals. 3. Establish: a) error rates, system operating curves, and error 
cost curves with different cost structures, together with their 95% confidence 
ranges for assessment of the classification performance of the flow cytometric 
approach to automated gynecologic prescreening, b) performance criteria against 
which other systems may be judged, c) benchmark data in terms of which goals of 
the cytology automation program may be assessed. 

Progress: We have established that flow cytometric measurement of cellular DNA 
content (as probed by chromomycin A3 fluorescence) and size (as probed by or- 
thogonal light scatter) is able to detect premalignant and malignant cells when 
present in the sample. Treating specimens briefly with DNAse and centrifugal 
elutriation markedly reduces the incidence of false alarms due to clumps of 
normal cells, damaged cells, and non-human cell artifacts giving signals simi- 
lar to those from abnormal cells. Sample diagnosis is based on the Atypia 
Index (AI) which is the ratio of Plain of Dysplasia signals (high fluorescence 
and moderate light scatter) to all epithelial cell signals. In a small pilot 
study, a specimen was classified as suspicious when its AI exceeded 0.5 per- 
cent. Comparison of machine classification with the cytomorphologic diagnosis 
indicated a false positive rate of 0.32 (12/33) and false negative rate of 0.09 
(3/31). 

The present study applies our protocol to greater numbers of specimens. 
Specimens from 85 healthy volunteers and from 80 women attending dysplasia 
clinic now have been collected and measured. This study already has demon- 
strated the feasibility of routinely measuring many samples. Analysis of the 
data will provide a statistically valid benchmark of the performance of flow 
cytometry in gynecologic prescreening. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



239 



CONTRACT RESEARCH SUMMARY 

Title: Imaging Improvement and Evaluation in Detecting Early Pancreatic Cancer 

Principal Investigator: Dr. A. R. Moossa 

Performing Organization: University of Chicago Hospitals 

City and State: Chicago, IL 

Contract Number: NOl-CB-84232 

Starting Date: 8/01/78 Expiration Date: 7/31/81 

Goal: To compare, improve, and evaluate imaging methods and strategies applied 
to early detection and diagnosis of pancreatic cancer. 

Approach: The imaging methods and strategies will be applied to the study of 
a selected population of patients in whom there is a high clinical probability 
of pancreatic cancer. The group will include patients with pathological condi- 
tions not related to the pancreas, patients with benign pancreatic disease, and 
patients with pancreatic cancer. The purpose of this study is to improve the 
ability of the diagnostic team to achieve rapid and certain identification of 
patients with early pancreatic cancer with the high probability group of pa- 
tients. Patients must be over 35 years of age and in suitable physical condi- 
tion to tolerate the necessary investigations and major abdominal surgery. 
They must have an estimated survival of at least three years. Both sexes will 
be included except for the exclusion of pregnant women. The study will be or- 
ganized into five separate projects. All the various imaging methods to be 
investigated will be included in one of the four projects. The fifth project 
will be concerned with collection and evaluation of data gathered by the other 
four projects. 

Progress: During the initial 30 months of the contract 113 patients with high 
clinical indications for probable pancreatic cancer have been entered into this 
study. Forty-four proved to have pancreatic cancer, 9 other cancers, 34 pan- 
creatitis and 26 had a normal pancreas. The combination of ultrasonography, 
endoscopic retrograde cholangiopancreatography (ERCP), computed tomography and 
arteriography continues to show promising results. Preliminary radionuclide 
scans with -'•-'•C- tryptophan are so much better than with ' ^Se-selenomethio- 
nine that -'--'■C-tryptophan produced by the local cyclotron and scanned with a 
special positron camera are now being supplemented by scanning with the Phocon 
(longitudinal multiplane emission tomography which was originally performed 
following selenomethionine injection) . Radionuclide scans are now performed 
both in fasting and in stimulated (after a meal; after injection of CCK-PZ or 
urocholine) state. Development of computer programs to produce a synthesized 
composite image by superimposing multiple images continues. The technical dif- 
ficulties of ERCP scanning following retrograde injection of technetium micro- 
spheres and of computerized ultrasound have now been sorted out. All of these 
approaches will be continued during the next six months of this study. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



« 



240 



» 



CONTRACT RESEARCH SUMMARY 

Title: Image Processing for Development of Automated Cell Recognition System 

Principal Investigator: Dr. George L. Wied 

Performing Organization: University of Chicago 

City and State: Chicago, IL 

Contract Number: NOl-CB-33873 

Starting Date: 5/31/73 Expiration Date: 6/30/81 

Goal: The total automation of clinical cytologic screening and diagnosis, 
focusing on definition of information requirements and criteria for high 
throughput, high resolution processing-based decision making. 

Approach: The contractor evaluated information requirements for the charac- 
terization and recognition by means of digital image processing of normal, pre- 
malignant and malignant cells from specimens derived from the human female 
genital tract, and for a subsequent classification into a diagnostic category. 
The high resolution system is complete and ready for intensive field testing 
on real-world cell data. 

Progress: Now fully automated (with the exceptions of automatically rotating 
the objective lens housing between medium and high resolution phases, and the 
automated insertion and removal of the specimen slide) the image processing and 
analytic system has been emendated by the addition of an improved scene segmen- 
tation technique using chromatic gradients to separate nuclei, cytoplasm and 
background, and cell classification performance has been significantly improved 
by the extraction of additional color and texture features, and Fourier shape 
descriptors calculated at high speed on the attached vector processor. Color 
images are now recorded on video tape, rather than being photographed, and 
zero-point and slide coordinate systems allow for quick relocation of cells on 
the original slide. 

Excellent performance over a variety of diagnoses has been achieved using 
specimens preselected for good preparation and staining, ample cytologic mate- 
rial, and relative freedom from contaminants and artifacts. To evaluate "real 
world" performance, it will be necessary to use routine specimens of average 
quality. Classification algorithms need to be adjusted to account for the in- 
creased variety of artifacts, contaminants and staining differences likely to 
be encountered in this broadened sample space. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



241 



CONTRA.CT RESEARCH SUMMARY 
Title: Statistical Center for Cooperative Lung Cancer Groups 



Principal Investigator: 
Performing Organization: 



City and State: 

Contract Number; 
Starting Date: 



NOl-CB-43868 
6/10/74 



Dr. C. Ralph Buncher 

University of Cincinnati Medical 

Center 
Cincinnati, OH 



Expiration Date: 5/31/82 



Goal: To collect and analyze information from the three clinical centers in 
the lung cancer screening program (which is directed toward the detection of 
early lung cancer in high-risk patients) to ascertain whether this screening 
program has reduced mortality and morbidity. 

Approach: Procedures have been established with each of the centers and agree- 
ment has been reached concerning the common data base for this study. Data are 
routinely monitored by the Central Statistical Group (CSG), translated into a 
single computer data base in Cincinnati, and analyzed to provide the combined 
collaborative information as well as comparative information. Reports are made 
quarterly. The CSG will continue to search the data from this study for impor- 
tant and statistically significant findings. 

Progress: The Cincinnati Central Statistical Group has established a common 
data base for this three clinical center study of screening to detect lung can- 
cer at an early enough time to reduce mortality. Translation and reporting 
systems for each of the clinical centers have been created and analyses of col- 
laborative results to date have been provided to the NCI and participating 
clinical centers. Intake of subjects has been completed so that adherence and 
compliance with the screening program is a current focus. Several thousand 
subjects have completed the screening program. Survival analysis efforts are 
being made by the CSG to obtain meaningful results with respect to mortality 
differences in the screening groups at as early a time as feasible. Regular 
Mortality Review Committee conferences are held to discuss the death certifi- 
cate and best Information causes of death for each man in the study who has 
died. Other central roles have been fulfilled by the CSG. In the first part 
of this contract, CSG was also responsible for a five-center study of computed 
tomography compared to radionuclide scanning and other methods of detection and 
diagnosis of brain tumors. Publications from that study have been issued. 
Similarly, CSG has worked with three centers concerning a collaborative study 
of pancreatic cancer diagnostic procedures; a number of reports from that study 
have been published. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $208,397 



242 



CONTRACT RESEARCH SUMMARY 

Title: Cryopreservation of Human Monocytes for Use in Immunologic Studies 

Principal Investigator: Dr. Roy S. Weiner 

Performing Organization: University of Florida 

City and State: Gainesville, FL 

Contract Number: NOl-CB-74131 

Starting Date: 9/30/77 Expiration Date: 12/31/80 

Goal: To isolate human monocytes from peripheral blood and develop methods for 
their adequate cryopreservation. 

Approach: Develop methods of collection and isolation of peripheral blood 
monocytes that would be applicable to normal subjects and to patients with can- 
cer. Develop optimum techniques for cryopreservation and define the functional 
characteristics of cryopreserved cells. 

Progress: To date, we have been successful in separating large numbers of 
monocytes from the peripheral blood using pheresis techniques. Up to 2 x 10^ 
monocytes can be collected in 60 ml of buffy coat from peripheral blood using 
a standard Haemonetics Model 30 blood cell separator. Further, we have devel- 
oped isolation techniques to obtain purified monocytes by elutriation centrifu- 
gation. We have modified techniques of Sanderson et al. so that up to 1.5 x 10^ 
mononuclear cells can be loaded into the elutriation chamber and up to 3 x 10^ 
monocytes are obtained in greater than 90% purity, representing 70% yield from 
peripheral blood. Additionally, we have separated a population of monocytes 
representing 15% of the total blood monocytes which are smaller than the large 
monocyte population. By fractionating the elutriation volume we were able to 
determine that these monocytes have a modal volume of 335 Ji^, whereas the 
large population of monocytes has a modal volume of 374 fi^ . We have also 
determined that while the native cytotoxicity attributed to monocytes is a 
function of the small population of monocytes, antibody cellular cytotoxicity 
is a function of both the small and large populations of monocytes. The anti- 
body dependent cellular cytotoxicity is masked in the small population of mono- 
cytes unless that population is further purified to avoid steric hindrance. 
We have shown that cryopreservation by standard techniques is satisfactory for 
the maintenance of hexose monophosphate shunt activity, native cytotoxicity, 
chemotaxis, and antibody dependent cellular cytotoxicity. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



243 



CONTRACT RESEARCH SUMMARY 

Title: Development of Contrast Agents for Use in Clinical Ultrasonic Diagnosis ^ I 

Principal Investigator: Dr. Allan H. Gobuty 

Performing Organization: University of Kansas Medical Center 

City and State: Kansas City, KS 

Contract Number: NOl-CB-84236 

Starting Date: 6/26/78 Expiration Date: 6/30/82 

Goal: To improve capability of ultrasound in the imaging of small tumors 
through the use of appropriate contrast agents. 

Approach: Chemical compounds having the appropriate physical properties to 
influence ultrasonic signals will be identified and equipment will be assembled 
to test such compounds. Differential uptake of these compounds will be mea- 
sured in animal tumor models. Animal toxicity studies will be made with pro- 
mising compounds. Clinical trials will be conducted using the most promising 
agents after appropriate toxicity studies have been completed. 

Progress: Compounds were selected and categorized ultrasonically by measuring 
the speed of sound, density and acoustic impedance as a function of concentra- 
tion in aqueous solution. Two groups of substances in solution, amino acids 
and chelates, have shown promise because of their ability to measurably alter 
both the speed of sound and acoustic impedance, and because they possess useful 
pharmacologic properties. After completing preliminary lethality studies on 
these compounds to assess their acceptability for intravenous use in animals, ^ 
one of them, Na2Ca EDTA, was infused into the jugular vein on an anesthetized 
dog. Enhancement of echoes of canine renal cortex was noted. The results ob- 
tained indicate that the chelating agent produced observable changes in scat- 
tering and reflection properties of renal cortical tissue. Moreover, as a 
group the chelates appear to be less toxic than compounds discussed in our 
previous progress report . 

Solid microspheres of collagen and gelatin, within the range of 2-10 ^M 
diameter, have also been prepared. Measurements of the ultrasonic backscatter 
from these in suspension and from a standard RBC preparation were performed. 
When normalized to a standard particle size and concentration, the back-scatter 
from collagen was about 30 dB above RBC. Because of the expenses involved in 
preparation of collagen spheres, comparison was made with the ultrasonic prop- 
erties of gelatin microspheres, both administered separately to anesthetized 
dogs. It was assumed that both these are taken up by RES cells. The canine 
experiments, using commercial diagnostic equipment (real-time linear array at 
7 MHz), demonstrated enhanced backscatter from the liver shortly after intra- 
venous infusion of each type of microsphere. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: 



244 



t 



CONTRACT RESEARCH SUMMARY 

Title: Purification of Human Tumor-associated Antigens 

Principal Investigator: Dr. David Gold 

Performing Organization: University of Kentucky Medical Center 

City and State: Lexington, KY 

Contract Number: NOl-CB-84259 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal: Purification, isolation, and identification of tumor-associated 
antigens. 

Approach: Extract and purify colonic mucoprotein antigen (CMA) from normal 
colon and colonic adenocarcinomas; develop radioimmunoassay and immunohisto- 
chemical procedures for detection of CMA; investigate the potential of CMA as 
a tissue or tumor-specific marker. 

Progress: This contract is concerned with the isolation of colonic mucoprotein 
antigen (CMA) and development of radioimmunoassay and immunohistochemical tech- 
niques for the detection of this organ specific antigen. During the first year 
emphasis was placed on purification of the antigen. A mucin fraction was ob- 
tained which by physicochemical criteria appeared to be relatively homogeneous. 
However, this material was found to be immunologically heterogeneous; two pre- 
cipitin arcs were observed in immunodiffusion with homologous antiserum. In 
the second year we have studied the nature and relationship of these two com- 
ponents. Efforts to separate these two materials intact (including molecular 
sieve, ion exchange, adsorption, affinity, and electrophoretic techniques) 
proved fruitless; however, by protease digestion followed by molecular sieve 
chromatography we were able to separate the two components to apparent homo- 
geneity, immunological as well as physicochemical. Analysis of these two mate- 
rials has indicated that they are both mucins, one a sialomucin and the other 
a fucomucin. They were shown to be immunologically distinct, and the organ 
specific determinant was found only in the sialomucin. Immunohistochemical 
procedures have identified the organ specific determinant in 60% (total of 59 
tumors examined) of colon carcinomas. Of particular note was the absence of 
the organ specific determinant in 14 gastric, 11 pancreatic, 9 endometrial and 
5 bronchogenic tumors, many of which were mucin producers. Assays for monitor- 
ing levels of antigen in body fluids are being developed. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



245 



CONTRACT RESEARCH SUMMARY 

Title: Immunohistochemical Studies of Tumor-associated Antigens 

Principal Investigator: Dr. F. James Primus 

Performing Organization: University of Kentucky Medical Center 

City and State: Lexington, KY 

Contract Number: NOl-CB-84257 

Starting Date: 9/18/78 Expiration Date: 9/17/81 

Goal: To isolate and characterize cancer-related markers in or on tumor cells 
by immunohistochemical techniques and describe their relationship to the dis- 
crimination of neoplastic cells from normal cells. 

Approach: Studies will be performed using colon carcinoma antigen III (CCA- 
III), colon-specific antigen protein (CSAp), fiHCG, Regan isoenzyme, and major 
blood group antigens in an attempt to discriminate between malignant and non- 
malignant cells and to determine whether immunocytochemical methods can accu- 
rately detect metastatic spread in regional nodes. 

Progress: Immunoenzyme staining for tumor antigens using glucose oxidase-anti- 
glucose oxidase (GAG) complexes gave similar localization and sensitivity as 
that obtained with PAP but without the inherent problems of endogenous enzjnne 
activity associated with the latter. The GAG and PAP methods were combined to 
localize CEA and CSAp simultaneously. Excellent contrast and staining separa- 
tion were shoxjn between the enzymatic reaction products of the two systems. 
Immunostaining with the GAG method of 84 histologically negative nodes from 15 
patients with CEA-positive primaries did not increase the incidence of tumor 
positive nodes. One of the latter nodes was positive for antigen localized 
within histiocytes. Immunostaining of a smaller number of histologically posi- 
tive nodes showed complete agreement except in one case in which the primary 
tumor was of borderline positivity. CEA positive staining was demonstrated in 
13 of 19 specimens of morphologically normal colonic mucosa reacted with the 
PAP method. Staining of normal colonic mucosa fixed in ethanol-acetic acid 
revealed a cytoplasmic localization of CEA in the columnar cell lining the 
mucosal surface and upper levels of the glandular crypts. CEA-specific locali- 
zation was not observed in colonic goblet cells or in the small intestine. 
Comparative studies of blood group antigen localization with the immunoenzyme 
technique and SRCA test have demonstrated the superiority of the former method 
in detecting blood group antigens in intestinal, squamous, and transitional 
epithelium. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



246 



CONTRACT RESEARCH SUMMARY 

Title: Screening Technique for Blood in Stool to Detect Early Cancer of Bowel 

Principal Investigator: Dr. Victor A. Gilbertsen 

Performing Organization: University of Minnesota Health 

Sciences Center 
City and State: Minneapolis, MN 

Contract Number: NOl-CB-53862 

Starting Date: 6/30/75 Expiration Date: 6/29/81 

Goal: To demonstrate a significant reduction in mortality due to colorectal 
cancer between the annually screened and the control group by employing the 
Hemoccult (R) form of the guaiac test for stool blood in combination with a 
diagnostic protocol for the source of bleeding. 

Approach: Forty-fiye thousand participants between the ages of 50 and 80 years 
residing in the state of Minnesota are randomly allocated to three groups on 
the basis of age, sex and geographic location in the state. Slides are com- 
pleted after the observance of a meat-free diet with suggested high fiber con- 
tent. One of the groups completes the slides once per year, another every 
other year, and a third group which is unscreened serves as the control. Each 
slide set consists of 6 slides representing two from each of three consecutive 
stools. Participants submitting samples positive for blood receive a diagnos- 
tic examination (including a colonoscopy, UGI series, and proctoscopy) to 
determine the cause of bleeding. All diagnostic procedures are conducted at 
the University of Minnesota Medical Center. All data including pathology 
reports on biopsy material and surgical specimens are computerized for easy 
retrieval. Extensive follow-up procedures are designed to retain participation 
of all subjects throughout the five years of testing and an additional five 
years of follow-up. Extensive dietary and health history data are collected 
from each subject via questionnaire. 

Progress: Significant progress to date includes completion of the first screen 
(both groups screened) and determination that 8% of the first screenees submit- 
ting at least 1 slide positive for occult blood and examined at the University 
of Minnesota (84%) were found to have at least one GI malignancy. The 16% 
examined elsewhere under a variety of protocols (most not including colono- 
scopy) have been accounted for and found to have a rate of 9% GI malignancies. 
The comparability of the two rates is being studied while the second and third 
screens are being completed. The diagnostic protocol for detection of the 
source of bleeding has been modified to exclude the single column B.E. x-ray 
subsequent to data indicating the superior uniformity of results using colono- 
scopic examination with air contrast B.E. x-ray (in those cases in which the 
cecum is not reached). Extensive follow-up procedures are followed to maximize 
continued participation and minimize attrition over the five-year screening and 
five-year follow-up periods. 



Project Officer: Mr. Louis P. Greenberg 
Program: Diagnosis 
FY 81 Funds: $604,000 



!47 



CONTRACT RESEARCH SUMMARY 

Title: Antigens on Human Lymphoid Organs: Immunodiagnosis of Leukemias and 
Lymphomas 

Principal Investigators: . Dr. John H. Kersey, Dr. Tucker W. 

LeBien 
Performing Organization: University of Minnesota 

City and State: Minneapolis, MN 

Contract Number: NOl-CB-84261 

Starting Date: 9/30/78 Expiration Date: 9/29/81 

Goal: To evaluate lymphoid differentiation antigens, particularly those on 
bone marrow and thymus cells, as potential immunodiagnostic markers for leuke- 
mia and lymphomas, and as markers for functional lymphoid cell populations. 
Approach: To produce monoclonal antibodies against antigens present on the 
human pre-B acute lymphoblastic leukemia cell line NALM-6 MI and antigens pres- 
ent on human T Ijnnphocytes. Use these monoclonal antibodies to a) examine the 
expression of the identified antigens on human leukemic cells, normal bone mar- 
row and normal thymus, and b) characterize the identified antigens using 
immunochemical analyses. 

Progress: Three new monoclonal antibodies have recently been produced and are 
undergoing extensive characterization. 

Monoclonal Antibody BA-1 : Produced by immunizing mice with the pre-B ALL 
cell line NALM-6-MI, BA-1 reacts with peripheral blood B cells, CLL, pre-B ALL, 
and most non-T, non-B ALL. BA-1 does not react with multiple myelomas, PWM- 
induced plasma cells, normal and malignant cells of T cell origin, monocytes, 
AML, AMML. The determinant recognized by BA-1 is not surface Ig, HLA-DR, Fc 
receptors, or C3 receptors. Immunochemical studies to date have shown that the 
cell surface antigen recognized by BA-1 is extremely sensitive to proteases. 
This characteristic has made it difficult to study BA-1 using radioimmune pre- 
cipitation and SDS-PAGE, and we are still working on determining the molecular 
weight of the BA-1 antigen. 

Monoclonal Antibody BA-2 : Also produced by immunizing mice with the 
NALM-6-MI cell line, BA-2 recognizes a 24,000 dalton protein (p24) in SDS-PAGE. 
p24 is found on 70% of non-T, non-B ALL (including most pre-B), and 3-6% of 
normal bone marrow cells. Double fluorochrome studies have shown that some 
BA-2+ bone marrow cells are also Tdr*". p24 is also expressed on 50% of AML 
and CLL, but is present on 2% of peripheral blood mononuclear cells. This 
newly described hematopoietic progenitor/leukemia-associated antigen is clearly 
different from the previously described common acute Ijnnphoblastic leukemia 
antigen (CALLA) , the latter being a glycoprotein of 100,000 daltons. 

Monoclonal Antibody TA-1 ; Produced by immunizing mice with the T ALL cell 
line HSB-2, TA-1 binds to all mature, peripheral T cells, 70% of thymocytes, 
and all peripheral blood monocytes. Studies with a F(ab')2 of TA-1 show that 
this monoclonal antibody is reactive with AMML, but nonreactive with AML. TA-1 
appears to react primarily with cells (malignant and nonmalignant) of T lympho- 
cyte or monocyte lineage. Recent studies using radioimmune precipitation and 
SDS-PAGE have shown that TA-1 precipitates a bimolecular complex of 175,000/ 
110,000 daltons. 

Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 

248 



« 



1 



CONTRACT RESEARCH SUMMARY 
Title: Slit-Scan Technique as Cancer Prescreening Automated System in Cytology- 
Principal Investigator: Dr. Leon L. Wheeless, Jr. 
Performing Organization: University of Rochester Medical 

Center 
City and State: Rochester, NY 

Contract Number: NOl-CB-33862 

Starting Date: 3/19/73 Expiration Date: 3/18/82 

Goal: Automation of clinical cytologic screening and diagnosis. 

Approach: The contractor will evaluate the performance characteristics of the 
X-Y-Z Slit-Scan Flow System. A benchmark study of the X-Y-Z system will be 
performed to (1) determine system characteristics including rate and causes of 
remaining false alarms and (2) document true alarm rates for abnormal speci- 
mens. Correlation studies will continue and studies on second stage processing 
techniques will begin. 

Progress: The X-Y-Z Slit-Scan Flow System has been fabricated and testing ini- 
tiated. A spectrum of normal and abnormal clinical specimans is being used to 
evaluate system performance characteristics and establish a flow data base. 
Preliminary results are very encouraging. Three hundred and seven clinical 
specimens have been analyzed to date. The false positive rate on 185 normal 
specimens is 12.3 percent. The false negative rate for abnormal specimens 
representing squamous cell cancer and its precursors is 4.2 percent. Out of 
the 76 abnormal specimens, only three slight dysplasias were missed. For the 
full spectrum of abnormality, the false negative rate is 7.2 percent. All 
false negatives were specimens having less than one abnormal cell per 1,000 
normal cells. 

An alternate system concept has been tested to provide similar multidimen- 
sional slit-scan information from a less complex instrument. This new instru- 
ment has been shown feasible and would additionally provide a significant 
increase in system throughput. The static cell data base has been expanded and 
specimen preparation protocols continue to be evaluated and improved. Specifi- 
cations for a cell sorter and second stage processor have been defined. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: ^359, 716 



249 



CONTRACT RESEARCH SUMMARY 

Title: Soluble Antigen- Antibody Complexes in the Circulation 

Principal Investigator: Dr. Harold J. Wanebo 

Performing Organization: University of Virginia Medical Center 

City and State: Charlottesville, VA 

Contract Number: COl-CB-84262 

Starting Date: 9/28/78 Expiration Date: 9/27/81 

Goal: Determine the diagnostic significance of circulating antigen-antibody 
complexes in the sera of cancer patients, identify the source of the antigen, 
and develop a sensitive assay for its presence. 

Approach: Patients in three tumor groups (breast, melanoma, colon) are being 
studied prospectively from presentation to determine the incidence and nature 
of immune complexes (IC) in serum and the changes in levels of IC that result 
from treatment. Immune complexes were identified by the presence of cryoglobu- 
lins and the Raji cell and Clq binding radioimmunoassays. 

Progress: In 24 patients with primary breast cancer, IC were detected in 39% 
at presentation with the Raji cell RIA, the most sensitive assay. There was a 
significant rise in Raji binding complexes two months after surgery (71%) fol- 
lowed by a fall. Patients with benign breast lesions had similar incidence of 
IC (35%) although levels were lower. Metastatic cancer was associated with a 
similar incidence of IC (57%). Preliminary data suggested that estrogen recep- 
tor positive tumors were associated with a higher incidence of circulating IC. 

In 18 primary melanoma patients, IC were detected at the onset in 25%. 
There was a similar increase, though less marked, in IC levels two months after 
surgery (46%). While there was a higher incidence of IC in patients with meta- 
static disease (57%), the absolute levels were not significantly higher than 
in those with primary disease. 

In 22 patients with colorectal cancer, IC were detected preoperatively by 
at least one assay in 64%, by the Raji RIA 59%, by Clq binding assay in 18% and 
by cryoglobulins in 5.3%. The frequency of IC by Raji assay was greatest in 
patients with Stage III disease (Dukes' C cancer). There were correlations of 
IC with CEA depending on the method of IC determination. With respect to Raji 
cell RIA assay in 61 patients who had CEA tests above normal, Raji cell values 
(exceeding 25 fig equivalent AHG/ml) were detected in only 18.7% (4 of 22) of 
patients with CEA values _> 20 ng/ml compared to 41% (16 of 39 patients) with 
CEA values <20 ng/ml, P = .02. Examination of Clq results showed positive Clq 
binding assays (values >7.5% precipitation) in 11 of 137 serum samples. Of 
these, 6 occurred with CEA values of 5-19 ng/ml and 5 occurred with CEA > 100 
ng/ml. Studies using sucrose density gradient ultracentrifugation and radio- 
immune diffusion showed that Clq was able to bind to CEA. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



250 



1 



CONTRACT RESEARCH SUMMARY 

Title: A Serum Alkaline Phosphatase Variant (FRAP) in Cancer Patients 

Principal Investigator: Dr. Frank C. Larson 

Performing Organization: University of Wisconsin 

City and State: Madison, WI 

Contract Number: NOl-CB-74173 

Starting Date: 9/15/77 Expiration Date: 12/14/80 

Goal: To define the diagnostic and prognostic significance of an isoenzyme of 
alkaline phosphatase (FHAP) in serum of human cancer patients. 

Approach: Serum of cancer patients often contains high level activity of a 
fast electrophoretically mobile, homoarginine sensitive alkaline phosphatase 
(FHAP), which'is present only at low activity in normal persons and those with 
benign disease. 

FHAP is ubiquitous among neoplastic diseases although some, e.g., carcinoma 
of the pancreas, lung and colon, are more likely associated than others, e.g., 
lymphomas. Initial studies were undertaken to examine the diagnostic implica- 
tion of FHAP. The studies during the final year focused on cancer prognosis. 

Progress: The positive relationship between serum FHAP activity and the pres- 
ence of cancer has been confirmed. Based on the analysis of several thousand 
persons' serum, an upper limit of normal value of 2.1 U/L has been established. 
Most cancer patients have values above 2.1 U/L. Highest values are more 
observed in patients with extensive disease. 

During the last year the research emphasis has been on comparing CEA and 
FHAP as tests of prognosis. Initially the approach was to compare FHAP values 
in patients whose prognosis was estimated to be good with those of patients 
whose prognosis was unfavorable. The mean (and median) value of patients who 
were judged to be cancer-free following surgical resection was substantially 
lower than the mean (and median) value of those who were believed or known to 
have cancer remaining. Of those with a favorable forecast, only 25% had values 
above normal (mean 1.86 U/L) while 62% of those with unfavorable forecast had 
above normal values (mean 9.27 U/L). 

During the third year of the contract sequential FHAP and CEA values were 
compared in 250+ patients to determine if changes in FHAP and CEA were predic- 
tive of recurrence and if so how long before the clinical evidence of relapse 
appeared. The most common event was that neither marker became elevated and 
the cancer did not recur during the period of study. In those cases in x-7hich 
cancer did recur in most cases either FHAP or CEA or both were increased. The 
increase in the cancer marker often antedated clinical evidence of recurrence 
as much as several months. FHAP appeared to be a somewhat more sensitive marker 
than CEA but, more importantly, the two markers seem to vary independently. 



Project Officer: Dr. Bernice T. Radovich 
Program: Diagnosis 
FY 81 Funds: 



251 



CONTRACT RESEARCH SUMMARY 

Title: Development of Large Area Solid State Image Receptors for X-Ray Imaging 

Principal Investigator: Dr. Moshe Eln-Gal 

Performing Organization: Xerox Corporation 

City and State: Pasadena, CA 

Contract Number: NOl-CB-74211 

Starting Date: 9/30/77 Expiration Date: 6/30/81 

Goal: To improve clinical diagnostic radiology and to provide for the diagno- 
sis of early cancer by developing a computerized imaging system based on a 
large area solid state x-ray receptor superior in clinical usefulness to those 
currently available. 

Approach: A large area solid state x-ray detector will be produced which will 
transform the x-ray flux into a digitally stored high quality image. The sys- 
tem to be developed, an x-ray selenium electronic linear scanner, will use the 
capability of a selenium alloy photoreceptor to create a latent image from 
absorbed x-rays. The latent image, a distribution of electric charge on the 
photoreceptor surface, will be detected by a microelectrometer scanning 
arrangement. Signals from the electrometer array will be coupled with the 
necessary control, display, and image storage memory components. Feasibility 
of the imaging system components will be developed in the first year. Further 
design, development, and optimization of the system will be completed during 
the second year. Clinical evaluation of the total imaging system will be con- 
ducted during the third year as well as further improvements and optimizations. 

Progress: Initial assembly and feasibility testing of the detection components 
were accomplished during the first year as scheduled. The computer system, the 
dedicated image processing hardware and the x-ray facilities were installed 
during the second year of the contract. Since the beginning of the third year, 
the system has been prepared for hospital installation, a cassette scanner has 
been designed and a high resolution (1024^) display unit has been fabricated. 
A team of radiologists for the clinical evaluation has been selected and the 
system was installed in the hospital June 1980. Imaging of live patients has 
begun. A cassette-oriented system has been developed and is undergoing physi- 
cal testing. It is expected to start clinical evaluation May 1981. 



Project Officer: Dr. Bill Bunnag 
Program: Diagnosis 
FY 81 Funds: $90,000 



« 



252 



CONTRACT RESEARCH SUiMMARY 

Title: Characterization of HLA Antigen of Donor's Lymphocytes by Serotyping and 
Cellular Typing 

Principal Investigator: Dr. Rene J. Duqesnoy 

Performing Organization: Blood Center of Southeastern Wisconsin 

City and State: Milwaukee, WI 

Contract Number: NOl-CB-04337 

Starting Date: 6/2/80 Expiration Date: 6/1/83 

Goal: To analyze as carefully as possible the cell surface histocompatibility 
antigens of donors in order to analyze the relationship between these antigens 
and the ability of those donors' cells to mount appropriate immune responses. 

Approach: Analysis of cell surface antigens is performed by two different 
detection systems: serology and cellular typing. The serologic analysis is per- 
formed using carefully screened alloantisera in assays of complement-dependent 
cytotoxicity. The cellular analysis is performed by analyzing the proliferative 
responses to homozygous typing cells (HTC typing) and by analyzing secondary 
restimulation of lymphocyte populations selectively immunized against alloantigens 
in primary response (PLT). 

Progress: As of 6/1/81 approximately 120 donors had been typed by serologic 
techniques and 20 donors by homozygous typing cell (HTC) analysis. The analysis 
of results with normal donors has allowed a definitive description of the popula- 
tion association between the SB markers of a new HLA locus (defined in the 
Immunology Branch) and the previously known HLA markers. This association is 
surprisingly weak, indirectly suggesting that the SB gene may map quite some dis- 
tance from the other HLA markers, and indicating that the SB markers will be use- 
ful new markers for population studies among normals and in disease. The results 
previously described in the summary of 12/2/80 regarding SB antigens in the 
disease Dermatitis Herpetiformis have been strengthened by studies of an additional 
13 donors. Family studies are now in progress to determine whether the evidence 
for involvement of both the DR and SB phenotypes in prediction of the disease 
results from gene interaction or from linkage disequilibrium. 

For further information see Annual Report #Z01-CB-O5067 I, ZOl-CB-05078 I, 
ZOl-CB-05100 I, and ZOl-CB-05101 I. 



Significance to Cancer Research: Considerable evidence from animal models and from 
epidemiologic studies in humans suggests that host cellular immune responses are 
crucial in determining the outcome of neoplastic diseases. Cellular immune 
responses are under control by genes in the major histocompatibility complex (HLA 
in man). In order to therapeutically manipulate these cellular immune responses, 
we must first understand their normal operation and genetic control. 

Project Officer: Dr. J. Stephen Shaw 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: $93,371 



253 



CONTRACT RESEAilCH SUMMARY 

Title: Maintain an Animal Holding Facility and Provide Attendant Research Services 

Principal Investigator: Ms. Diana Hernandez 

Performing Organization: Cor Bel Laboratories, Inc. 

City and State: Rockville, MD 

Contract Number: NOl-CB-04336 

Starting Date: 11/1/79 Expiration Date: 10/31/82 

Goal: Maintain colonies of inbred mice (10,000 animals), inbred rats (500 animals), 
and rabbits (20 animals) and carry out selected breeding protocols with these ani- 
mals as specified by the project officer. These animals are to be maintained in 
support of intramural research programs in the Immunology Branch, NCI. , 

Approach: Colonies of mice, rats, and rabbits are to be housed and fed according 
to National Research Council standards. Technical manipulations and breeding are 
to be carried out as directed by the project officer. 

Progress: Performance on this contract has been highly satisfactory. The animal 
colonies have been established and are being maintained according to National 
Research Council standards. Animal health has, in general, been excellent, and 
breeding protocols have been satisfactory. Recordkeeping and transferring of 
animals to and from the NIH Campus have all been satisfactory. j 



d 



Significance to Cancer Research: This animal colony is necessary in support of 
intramural research programs in the Immunology Branch of NCI. Many of these 
programs are concerned with the immune response to cancer. 

Project Officer: Dr. David H. Sachs 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: $295,500 ($68,500 estimated additional) ,^| 

254 



CONTRACT RESEARCH SUMMARY 

Title: Study of Natural Cellular Immunity to Tumors in Mice and Rats 

Principal Investigator: Mr. Brian Weatherly 

Performing Organization: Cor Bel Laboratories, Inc. 

City and State: Rockville, MD 

Contract Number: NOl-CB-14346 

Starting Date: 2/16/81 Expiration Date: 2/15/84 

Goal: Study of natural cell-mediated cytotoxicity of mice and rats and/or 
their immune responses to tumor-associated antigens. Studies on in vivo resis- 
tance to tumors and correlation with in vitro cellular immune responses. 

Approach: To study (1) the natural cell-mediated cytotoxicity against tumors 
of mice and rats, and (2) the factors influencing the levels of this reactivity 
and mechanism of cytotoxicity. 

Progress: This project was just initiated by this contractor, after recompeti- 
tion of the previous contract (NOl-CB-74093) . The necessary training of per- 
sonnel, transfer of animals, and setting up of techniques for _in vivo experi- 
mentation have gone well and have been completed. 

Mice which received leukemogenic doses of x-irradiation continue to be 
followed for the development of thymic lymphomas. Some mice have been inocu- 
lated with bone marrow from normal or irradiated mice to determine the effects 
on tumor development and on NK cell activity. 

Breeding studies are being continued, to place the genes for low NK activ- 
ity on other genetic backgrounds. 



Project Officer: Dr. Ronald B. Herberman 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: $222,228 

255 



CONTRACT RESEARCH SUMMARY 

Title: Administrative Support Services 

Principal Investigator: Ms. Karen Meinzen 

Performing Organization: CSR, Inc. 

City and State: Washington, D.C. 

Contract Number: 263-79-C-0021 

Starting Date: 10/19/80 Expiration Date: 10/18/81 

Goal: To support administrative staff in preparation of scientific meetings 
and the research components in accomplishing their administrative tasks. 

Approach: Current staffing situation places excessive demands on the admini- 
strative staff in carrying out their responsibilities. This project is to 
support tasks in organizing overview meetings, scientific workshops, and in 
preparing pre- and post-conference materials. In addition, editorial support 
was available to researchers in satisfying specific demands and formats of 
scientific journals. 

Progress: Ten conferences of standing committees and specially organized work- 
shops were held. Excellent support was given to NIH conference on low level 
radiation, in preparation of post-conference materials. Four-hundred eighty 
publications of current research were edited to correspond to requirements of 
specific journals. Pre-conf erence activities were handled for a number of 
meetings. On-site support was provided to workshops and overviews. Seventy- 
five hundred reprints were handled through the storage/distribution facility. 
Invaluable assistance was provided in preparation of annual reports and program 
overview handbooks. 



Project Officer: Ihor J. Masnyk, Ph.D. 
Program: DCBD 

Technical Review Group: Ad Hoc Committees 
FY 81 Funds: $382,775 



256 



CONTRACT RESEARCH SUMMARY 



Title: Computer Services 

Principal Investigator: 
Name/Address : 
Performing 
Organization: 

Contract Number: YOl-CB- 90316 
Starting Date: Nov. 15, 1979 



Mr. Francis A. McDonough 
Department of the Treasury 
1435 G Street, N.W. 
Washington, D.C. 20220 



Expiration Date: Sept. 30, 1981 



Goal : To provide computer facility for mathematical computations related to 
biological systems modeling carried out in the Laboratory of l^tathematical Biology 
and other groups in DCBD, NCI. 



Approach : Facility is accessed through remote terminals, 



Progress : This facility is used only in a backup capacity because our own VAX 
computer is taking over most of our computations. 



Project Officer: Dr. Mones Berman 
Program: Cancer Biology Support 
Technical Review Group: Ad Hoc 
Relevance Review Group: 
FY 81 Funds: $2,000. 



Site Visit Date: N/A 



257 



CONTRACT RESEARCH SUMMARY 



Title: A Screening System for Topical Chemotherapy of Mycosis Fungoides 

Principal Investigator: Dr. Stanford Lamberg 

Name/Address Johns Hopkins University 

Performing Organization: Baltim-ore, MD . 

Contract Number: NOl-CB-63927 

Starting Date: 6/30/76 Expiration Date: June 30,1981 

Goal: To apply chemotherapeutic agents developed by DCT for topical treatment 
of mycosis fungoides, a cutaneous lymphoma. 

Approach: Chemotherapeutic agents are obtained commercially and from the DCT, 
NCI, and these agents are being screened by clinical patch testing in patients 
with mycosis fungoides. 

Progress: Investigative new drug applications have been obtained fromt the FDA 
and patch test screening of these agents has been performed at Hopkins and other 
affiliated institutions comprising the Mycosis Fungoides National Cooperative 
Group. Patch test of all available chemotherapeutic agents has recently been 
completed . 



Significance for Cancer Research: (NCP Objective 6 Approach 2) 

Project Officer: Dr. Gary L. Peck 
Program: Biology Support 
Technical Review Group: Ad Hoc Committee 
FY 80 Funds : None 



258 



1 



CONTRACT RESEARCH SUMMARY 

Title: Melanoma Cell Vaccine in In Vitro Assays for Humoral and Cellular 
Cytotoxicity 

Principal Investigator: Dr. Grace Cannon 

Performing Organization: Litton Bionetics, Inc. 

City and State: Kensington, MD 

Contract Number: NO'l-CB- 53916 

Starting Date: 5/10/81 Expiration Date: 10/9/81 

Goal: To prepare cultured melanoma cells for immunotherapy vaccine and to harvest 
and store patient serum samples. 

Approach: (1) Grow human melanoma cells free of contaminants, treat cells with 
neuraminidase, freeze under sterile conditions, and provide cells as needed for 
human immunotherapy. (2) Deliver patient blood samples to contract site, harvest 
serum samples, and store them at -70°C. 

Progress: Since the previous contract research summary, 4 new batches of vaccine 
have been prepared from human melanoma cell lines UCLASM-6, UCLASM-12, and 
UCLASM-14. Vaccine preparations have met quality control tests. In addition, 58 
serum samples were frozen and stored in 657 vials. No additional vaccine produc- 
tion is needed to finish the protocol. Litton Bionetics staff will continue to 
deliver vaccine to the NIH Clinical Center until the contract expiration date. 



Significance to Cancer Research: The melanoma cell line vaccine will be used in 
a clinical immunotherapy trial. 

Project Officer: Dr. John R. Wunderlich 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: $0 



259 



CONTRACT RESEARCH SUMMARY 

Title: Studies of the Immune Response of Mice and Rats to Tumor Antigens " 

Principal Investigator: Dr. Grace Cannon 

Performing Organization: Litton Bionetics, Inc. 

City and State: Kensington, MD 

Contract Number: NOl-CB-74093 

Starting Date: 2/28/77 Expiration Date: 2/27/81 

Goal: Study of natural cell-mediated cytotoxicity of mice and rats and of their 
immune responses to tumor-associated antigens. Studies on in vivo resistance to 
tumors and correlation with in vitro cellular immune responses. 

Approach: To study (1) the natural cell-mediated cytotoxicity against tumors of 
mice and rats, and (2) the factors influencing the- levels of this reactivity and 
mechanism of cytotoxicity. 

Progress: The main emphasis has been on the role of natural killer (NK) cells 
in resistance against tumor growth in^ vivo . Isotopically labeled tumor cells 
were injected intravenously and their rate of clearance from the lungs and other 
organs assessed during the first 2-18 hrs. Clearance of the labeled tumor cells 
correlated well with known levels of NK activity. Mice given the carcinogen, 
ure thane, were analyzed in regard to levels of in vivo cytotoxic activity during 
the latent period. Mice which were susceptible to carcinogenesis also showed 
transiently depressed NK activity and depressed in_ vivo clearance of labeled ^ 
tumor cells^ whereas strains of mice that were resistant to carcinogenesis did ^ 
not show depressed reactivity. The carcinogenesis by ure thane in susceptible 
mice could be diminished by administration of normal bone marrow cells at 7-14 
days after the carcinogen, indicating that depression of host resistance may be 
a critical factor in tiamor induction. The possible role of NK cells in radia- 
tion carcinogenesis also is being examined. Fractionated leukemogenic doses of 
x-irradiation were found to markedly inhibit NK activity of C57BL/6 mice and 
this depression could be reversed by spleen or bone marrow cells from normal 
syngeneic mice but not from beige mice. Preliminary results also indicate a 
higher tumor incidence in irradiated beige mice. 

Extensive mouse breeding studies have been performed, to follow up on obser- 
vations regarding the genetic basis for low NK activity in the A and SJL strains. 
It appears, from F2 and backcross analyses, that one gene is responsible for the 
depressed reactivity and efforts are being made to place this gene on other ge- 
netic backgrounds, to facilitate studies on the role of NK cells in primary car- 
cinogenesis. 

Significance to Cancer Research: Understanding the relationship of in vitro 
assays to host resistance of tumors is an essential element in our ability to use 
these assays to monitor the clinical response to immunotherapy and to understand 
the role of various immune mechanisms in resistance to tumor growth. Recent find- 
ings with natural cell-mediated immunity may account for a significant portion of 
in vivo resistance to tumor growth, and may have important implications for immune 
surveillance. 



Project Officer: Dr. Ronald B. Herberman 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: 

250 



«! 



CONTRACT RESEARCH SUMMARY 

Title: Maintenance and Development of Inbred and Congenic Resistant Mouse Strains 

Principal Investigator: Ms, Martha McGowan 

Performing Organization: Litton Bionetics, Inc. 

City and State: Kensington, MD 

Contract Number: NOl-CB-94325 

Starting Date: 2/1/79 Expiration Date: 1/31/82 

Goal: To maintain a colony of inbred pedigreed strains of mice which are needed 
to support ongoing NCI intramural research in transplantation immunology. 

Approach: The contractor maintains a colony of approximately 40 special inbred 
and congenic resistant strains of mice by pedigreed brother-sister mating. Quality 
control testing is carried out at each generation by cytotoxicity typing of ani- 
mals from each strain. Alloantisera are raised between mouse strains to assist in 
this quaity control typing, and sera and animals are shipped by the contractor to 
collaborating investigators at NIH and elsewhere. 

Progress: The contractor has maintained all inbred and congenic resistant strains 
of mice in excellent condition. Breeding of each strain and of hybrid strains, 
recordkeeping, and quality control testing have all been highly satisfactory. A 
backcrossing program has been instituted for all congenic resistant strains in 
order to keep the backgrounds of these strains identical. This involves backcross- 
ing of each congenic to the reference background line once every 6-10 generations. 
This program has also been very satisfactory to date. Two new recombinant H-2 
haplotypes have been identified during the process of this backcrossing, and these 
have been bred to homozygosity and established as two new valuable inbred congenic 
strains. 

Antisera for histocompatibility antigen typing have been prepared in a variety 
of combinations and have been found to be excellent reagents. A series of new 
strain-restricted typing sera have been produced in order to identify each strain 
in the colony and distinguish it from all other strains. Shipping of animals and 
sera to collaborating investigators at NIH and elsewhere has been very satisfactory. 
The animals shipped from these pedigreed colonies have generally been of excellent 
health and have provided breeding stock for the production of larger numbers of 
experimental animals in numerous laboratories. 



Significance to Cancer Research: This animal facility is needed for the breeding 
and maintenance of these inbred congenic resistant strains of mice. These animals 
make possible research on individual histocompatibility antigens and, in particular, 
the role of the major histocompatibility complex in the transplantation of tissues 
and cells and in the immune response to cancer. 

Project Officer: Dr. David H. Sachs 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 80 Funds: $256,763 



261 



CONTRACT RESEARCH SUMMARY 



Title: Induction, Transplantation, and Preservation of Plasma Cell Tumors 
in Mice and the Maintenance of Special Strains 

Principal Investigator: Martha J. McGowan 

Performing Organization: Litton Bionetics, Inc. 

City and State: Bethesda, MD 

Contract Number: NOl-CB-94326 

Starting Date: 3/1/79 Expiration Date: 1/31/82 

Goal: Transplantation, preservation induction, and shipping of plasmacytomas, 
T- and B-cell lymphomas in mice. Breeding of rare strains of mice, hybrids, 
and wild mice. 

Approach: To maintain a closed conventional colony of BALB/c, BALB/c sublines, 
and BALB/c congenic strains for use in plasmacytoma induction and for the 
maintenance of a bank of transplantable plasmacytomas and other lymphocytic 
tumors. The mice in this environment are suitable for long-term plasmacytoma 
induction studies and for the generation of congenic strains of the BALB/c 
background. New BALB/c congenic lines, carrying genes and chromosomal segments 
from the plasmacytomagenesis-resistant DBA/2 C57BL, C3H and CBA backgrounds 
are being bred for use in plasmacytoma induction studies to identify genes involved 
with the genetic susceptibility of plasmacytomagenesis. In addition, to maintain 
a screening bank of myeloma proteins for the detection of new antigen binding 
myeloma and to maintain a wild mouse colony. 

Progress; The contractor has maintained a bank containing roughly 2000 frozen 
tumor lines. This bank is a source of standard plasmacytomas, lymphocytic 
tumors, mast cell tumors and histiocytomas that are shipped to other 
investigators. Approximately 400 shipments are made per year. 

The contractor maintains 47 different inbred strains, 31 wild stocks, and is 
developing 48 congenic strains. These mice are used to produce animals for 
plasmacytomagenesis studies. Currently the contractor maintains approximately 
19 studies involving 2868 mice. The wild mouse colony is now probably the 
largest in the U.S. It continues to provide an abundant source of new genotypes. 

Significance to Cancer Research: Provides essential support for the study 
of plasmacytomagenesis (carcinogenesis) with the specific goal of determining 
the genetic basis of susceptibility to tumor induction by mineral oil. 
Supplies essential biological material for investigators studying the biology 
of neoplastic plasma cells, tumor immunology, the genetics of immunoglobulins, 
and immunoglobulin synthesis. 

Project Officer: Dr. Michael Potter 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: $454,631 



26 2 



CONTRACT RESEARCH SUMMARY 

Title: Radioimmunoassay of Immunoglobulin Molecules 

Principal Investigator: Dr. James Harness 

Performing Organization: Meloy Laboratories, Inc. 

City and State: Springfield, Virginia 

Contract Number: NOl-CB-63932 

Starting Date: 3/14/81 Expiration Date: 3/13/32 

Goal: To perform radioimmunoassays of immunoglobulin molecules IgM, IgA, IgG, 
IgD, and IgE in lymphocytes culture supernatants or in other biological 
fluids . 

Approach: The contractor is to perform determinations of human IgG, IgA, 
IgM, IgD, and IgE, and mouse IgA and IgM by double antibody radioimmunoassay 
by procedures defined by the project officer and using reagents supplied by 
the project officer. This contract provides critically required research 
support for studies on the nature of immunodeficiencies that are associated 
with a high incidence of malignancy and on the cause of the immunodeficiency 
associated with malignancies of the B cell or plasma cell system. In 
addition, these studies are directed at defining retained immunological 
capabilities of T-cell leukemias . 

Progress: The contract has established radioimmunoassays for IgG, IgA, 
IgM, and IgE of man and for IgA and IgM of the mouse. These assays have 
been used to quantitate the rate of immunoglobulin synthesis by poksweed 
mitogen-stimulated, peripheral blood lymphocytes of man or of splenic 
lymphocytes of the mouse in in vitro cultures. Patients with a T-cell 
leukemia associated with the Sezary syndrome have been shown to have a 
malignant expansion of helper T cells whereas a patient with acute lymphocytic 
leukemia was shown to have a malignancy of the suppressor T cells. A subset 
of patients with common variable hypogammaglobulinemia has been shown to 
have excessive numbers of suppressor T cells that inhibit gamma globulin 
synthesis by B cells. Certain patients with post marrow transplant immuno- 
deficiency have deficient B-cell function whereas others have excessive 
suppressor T-cell activity. Patients with multiple myeloma have 
hypogammglobulinemia due in part to excessive numbers of suppressor 
macrophages. These studies are defining the nature of disorders of the 
immune system that led to a high incidence of cancer. In addition, they 
provide information on the cause of immunodeficiency that arises secondary 
to certain forms of malignancy. Finally, these studies have provided 
insights into the retained functions of T-cell leukemias. 

Significance to Cancer Research: These studies will help elucidate the 
abnormalities of the immune system associated with the development of cancer. 

Project Officer: Thomas A. Waldmann, M.D. 

Program: Cancer Biology Support 

Technical Review Group: Ad Hoc Review Group 
FY 81 Funds: $51,000 



263 



CONTRACT RESEARCH SUMMARY 

Title: National Cancer Institute Immunodiagnostic Reference Center 

Principal Investigator: Dr. James Harness 

Performing Organization: Meloy Laboratories, Inc. 

City and State: Springfield, VA 

Contract Number: NOl-CB-63976 

Starting Date: 7/11/80 Expiration Date: 7/10/81 

Goal: To develop new assays for tumor-associated antigens, to use immuno- 
chemical techniques for the diagnosis of cancer, to evaluate humoral immune 
responsiveness of patients with cancer. 

Approach: To use sensitive radioimmunoassays to quantitate human, monkey, 
and mouse alpha-fetoprotein and human chorionic gonadotropin in serum, to 
evaluate these assays both as diagnostic techniques for testicular, 
ovarian, and hepatocellular cancer, and as a means of monitoring the 
clinical response to therapy. The contractor performed tests of humoral 
competence of patients with cancer or immunodeficiency diseases. 
Solid phase radioimmunoassays and ELISA assays hve been established to 
measure antibodies produced by antigen stimulated human peripheral blood 
mononuclear cells. 

Progress: In a wide-ranging study of 1,000 patients with nonseminomatous 
germ cell tumors of the testis or hepatocellular tumors, alpha-fetoprotein 
and HCG assays have been shown to be of great value as an adjunct to diagnosis, 
in staging, and as a tool to monitor the chemotherpay of these tumors. These 
assays are being performed in collaboration with eight groups in monitoring 
the treatment of patients with hepatocellular or testicular germ cell 
tumors other than seminoma. In all cases in which the marker was elevated, 
but where there was no clinical evidence of disease, a positive marker always 
reflected occult residual tumor. Humoral antibody assays have been 
established for ten antigens and these antibody assays are of critical 
importance in defining immune defects in patients with high incidence of 
neoplasia. The solid phase radioimmunoassays and ELISA assays for hximan 
antibodies produced by antigen stimulated human peripheral blood mononuclear 
cells have been used to identify the pathogenic defects that lead to primary 
immunodeficiency diseases associated with an increased incidence of malignancy. 
Disorders involving T helper cell deficiency, excessive suppressor T cell activity 
and B cells have been identified in these patients with primary immunodeficiency 
states. 

Significance to Cancer Research: These studies are helping to establish 
the usefulness of these multiple immunodiagnostic tests in the diagnosis 
of cancer, or as a means of monitoring patients during and after therapy. 

Project Officer: Dr. Thomas A. Waldmann 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: 



264 



CONTRACT RESEARCH SUMMARY 



Title : Molecular Biologic Studies of Tumor Viruses 



Principal Investigator: 

Name/Address 

Performing Organization : 

Contract Number: NOl-CB-04342 
Starting Date: 6-30-80 



Dr. Richard S. Howk 
Meloy Laboratories 
Rockville, MD 



Expiration Date: 6-29-83 



Goal: To provide support for studies of the structure and expression 
of tumor viruses. 

Approach: Viral biochemical; serolgoic, and biological parameters 

are monitored in different systems to define the regulatory mechanisms 

of tumor virus expression and to relate the expression to tumor production. 

Progress : Mouse cells transformed by Bovine papilloma virus or by 
viral DNA have been found to contain multiple unintegrated viral DNA 
copies in the absence of detectable integrated viral DNA. These results 
suggest that transformation in this system is induced and maintained 
by unintegrated viral DNA. 

Efficient transformation by the p21 coding region of Harvey murine 
sarcoma virus (Ha-MuSV) DNA requires that the viral long terminal repeat 
(LTR) be ligated to the p21 coding region. Two independent p21 coding 
genes have been cloned from normal rat cell DNA; one gene is colinear 
with the viral p21 coding region, while the second gene contains 
intervening sequences. When the Ha-MuSV LTR is ligated to either gene, 
it can transform NIH 3T3 cellls , inducing high intracellular p21 levels. 

The origin and formation of mink cytopathic focus-forming (MCF) murine 
leukemia virus (MuLV) has been studied. Endogenous MCF-like viral DNAs 
have been found in AKR and other mouse strains. The entire env of some 
non-pathogenic MCF viruses has apparently been derived from these sequences 
In spontaneous murine thymic tumors, one or more of these MCF-like 
sequences have regularly recombined with a specific region of the 3' 
end of ecotropic env . Pathogenic MCF viruses also contain these ecotropic 
virus-derived recombinant sequences. 



Significance for Cancer Research ( NCP Objective 6 Approach 2) 

Project Officer: Dr. Douglas R. Lowy 
Assistant Project Officer: Dr. Ira H. Pastan 
Program: Immunology Support 
Technical Review Group: Ad Hoc Committee 
FY 8 1 Funds: $240,000 



265 



CONTRA.CT RESEARCH SUMMARY 

Title: Immune Status of Patients Undergoing Immunotherapy 

Principal Investigator: Dr. Douglas M. Strong 

Performing Organization: Naval Medical Research Institute 

City and State: _ Bethesda, MD 

Contract Number: YOl-DB-00319 

Starting Date: 3/1/80 Expiration Date: 9/30/82 

Goal: To study the immune responses of cancer patients to tumor-associated 
antigens and to provide support for the Laboratory of Immunodiagnosis, NCI, for 
its detailed studies of cell-mediated immunity to human cancer. 

Approach: Immunological tests to evaluate and monitor the immunocompetence of 
patients and their response to tumor-associated antigens include lymphocyte 
stimulation by mitogens and autologous tumor antigens; rosette assays for enu- 
meration of T cells; leukocyte migration inhibition assays against recall anti- 
gens and tumor-associated antigens; and studies of direct cytotoxicity and 
antibody-dependent cell-mediated cytotoxicity against various human tumor cell 
lines. 

Progress: All of the assays for cell-mediated immunity, and the computer sys- 
tem for analyzing the data and correlating with clinical information, have been 
fully implemented and are being performed well. Alterations in cellular immune 
function are being analyzed in cancer patients who are receiving BCG or poly 
I:C, or who are being treated by passage of their plasma over Staphylococcus 
aureus-protein A columns. The changes related to these treatments are under 
current analysis. 

During the past several months, a major emphasis has been placed on the 
monitoring of cancer patients receiving various doses of purified recombinant 
leukocyte interferon. Natural killer cell activity and several other immuno- 
logic parameters are being tested at frequent intervals. 

Studies on proliferation of human T cells in the presence of T cell growth 
factor have been pursued and techniques were developed for consistent produc- 
tion of high levels of factor, by stimulation of normal lymphoid cells and cer- 
tain B cell lines. 



Significance to Cancer Research: These studies will provide important informa- 
tion regarding the usefulness of various assays of cell-mediated immunity for 
immunodiagnosis and for monitoring the clinical course of cancer patients. 
They should also provide the necessary data for determining the optional dose 
of various agents for biologic response modification. 

Project Officer: Dr. Ronald B. Herberman 

Program: Tumor Immunology Program 

Technical Review Group: Immunology Support Contract (Ad Hoc Review) 

FY 81 Funds: $285, 181 

265 



CONTRACT RESEARCH SUMMARY 

Title: A Study of Phylogenetic Aspects of Neoplasia 

Principal Investigator: Dr. John C. Harshbarger 

Name/Address The Smithsonian Institution 

Performing Organization: Washington, D.C. 

Contract Number: NOl-CB-33874 

Starting Date: 7/1/73 Expiration Date: 6/30/82 

Goal: To collect, examine, classify, and preserve neoplasms in cold-blooded 
vertebrate and invertebrate animals, and to study experimentally the develop- 
ment of tumors in lower animals. 

Approach: The principal investigator directs the operation of a registry of 
tumors in lower animals. Specimens are acquired from personal field investi- 
gations or through submittal by other investigators. The specimens are examined 
grossly, histologically, and in some cases by electron microscopy. Diagnoses 
are established and the specimens are described. Field investigations and 
experimental inductions of tiimors in lower animals are carried out. Publications 
of the findings are made. The Registry also serves in a consulting capacity to 
other agencies concerned with diseases in lower animals. A complete collection 
of the literature on neoplasms in ectothermic animals is maintained on computer 
tape and is available for retrieval. Data on specimen accessions is likewise 
stored in a computer system. 

Progress: Specimen accessions during the past year numbered 152, bringing the 
total to 2371. The literature collection has been re-examined and irrelevant 
papers deteted, leaving 3755. Data from these have been stored on computer 
tape and are retrievable by author, species, organ, diagnosis, and other key 
words. About half of the new accessions were confirmed to be neoplasms, while 
the remainder were parasitic, infectious, toxic, traumatic, or developmental 
diseases. Neoplasms were mostly from teleost fishes, amphibians, reptiles, 
and molluscs. In the invertebrate phyla, lesions that appear to be neoplasms 
by morphological criteria were found only in molluscs and arthropods (insects 
alone represented) with the possible exception of one platyhelminth. About 
50 types of neoplasms have been recognized to occur at relatively high pre- 
valences in certain species in certain locations. Foreign contributions came 
from Canada, England, Australia, India, Germany, Netherlands, Egypt, Japan, and 
USSR. 

Significance for Cancer Research: Field studies and anatomical studies indi- 
cate environmental carcinogens that may be of importance in human cancer epi- 
demiology or may be useful in designing analytical experiments to determine 
mechanisms of tumorigenesis. 

Project Officer: Clyde J. Dawe, M.D. 

Program: Cancer Biology Support 

FY 81 Funds: 180,200 Site Visit Date: No Site Visit 



267 



CONTRACT RESEARCH SUMMARY 

Title: Preparation of Purified Wheat Proteins and Wheat Protein Fractions ^ 

Principal Investigator: Dr. Donald Kasarda 

Performing Organization: U.S. Department of Agriculture 

City and State: Berkeley, California 

Contract Number: YOl-CB-60312 

Starting Date: 10/1/80 Expiration Date: 9/30/81 

Goal: To obtain chemically defined fraction of wheat gliadin for use in 
studies of gluten-sensitive enteropathy (coeliac sprue) . 

Approach: Wheat gluten will be chemically fractionated and subjected to 
cyanogen bromide cleavage. Homogeneous fragments of gliadin, as well as 
gliadin itself, will then be supplied. 

Progress: The contractor has supplied alpha-gliadin according to the 
contract schedule. The alpha-gliadin has been used in _in vitro studies of 
responses of lymphoid cells to defined gliadin preparations — cells obtained 
from both patients and normals. Finally, alpha-gliadin has been used in a 
radiometric assay of anti-gliadin production by lympoid cells ixi_ vitro 
which will ultimately enable us to study the regulation of anti-gliadin 
responses in vitro . 

Significance to Cancer Research: Gluten-sensitive enteropathy is a disease 
associated with a high incidence of malignancy. Elucidation of the ^| 

pathogenesis of gluten-sensitive enteropathy will provide insight into the ^P 

factors which lead to the onset of malignant disease. 

Project Officer: Warren Strober, M.D. 

Program: Cancer Biology Support 

Technical Review Group: Breast Cancer Task Force Committee 
FY 81 Funds: $18,600 



m 



268 



CONTRACT RESEARCH SUMMARY 



Title: Preparation and Analysis of Cell Surface Protein (CSP) Fractions 

Principal Investigator: Dr. David Schlesinger 

Performing Organization: University of Illinois 

City and State: Chicago, IL 

Contract Number: NOl-CB-74214 

Starting Date: 9/30/80 Expiration Date: 9/29/81 

Goal: To prepare and analyze the structure of cell surface protein. 

Approach: The C-terminal fragment of CSP (fibronectin) is prepared by 
proteolytic cleavage, anion exchange chromatography, and high pressure 
liquid chromatography. Fragments of fibronectin are prepared by cleavage 
by trypsin, chymotrypsin, or thermolysin, followed by affinity chromatography 
on specific ligands such as collagen, actin, or heparin, followed by ion 
exchange or molecular sieve chromatography. The fragments are sequenced 
by sequenator or micro-sequencing protocols. 

Progress: Sufficient amino acid sequence has been obtained from a C-terminal 
fragment of fibronectin to be able to compare with recombinant DNA information 
from another project. A routine protocol was established for rapid amino acid 
analysis and sequencing of small amounts of purified fragments, and primary 
sequence data has been obtained from four entirely new fibronectin fragments, 
each of which is unique in activity and sequence. Further primary structure 
analysis is needed to provide an understanding of CSP's multiple biological 
effects. However, the Investigator who initiated the project is now leaving 
the contractor, and sufficient expertise to continue the project is no longer 
available at the University of Illinois. This particular contract will 
therefore be terminated at the end of the contract year. 

Significance to Cancer Research: CSP (fibronectin) is a major adhesive 
glycoprotein that is present in a variety of normal connective tissue and 
epithelial cell types and is absent from many types of cancer cells, 
especially metastatic cells. Knowledge of the structure of CSP will further 
the understanding of the cancer process. 



Project Officer: Dr. Kenneth Yamada 
Program: Cancer Biology Support 
Technical Review Group: Ad Hoc Committee 
FY 81 Funds: $25,942 



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269 



CONTRACT RESEARCH SUMMARY 

Title: Interaction of Exercise, Dietary Carbohydrate and Cancer Cachexia 
in Rats 

Principal Investigator: Dr. Richard A. Ahrens 

Performing Organization: University of Maryland 

City and State: College Park, MD 

Contract Number: NOl-CB-94327 

Starting Date: 6/1/79 Expiration Date: 5/31/82 

Goal: To investigate the separate and interactive effects of quantitatively 
imposed exercise and variation in dietary carbohydrate source on the systemic 
response of rats to growth of tumors. 

Approach: The first phase of the project will consist of determining optimum 
ranges of exercise schedule and dietary carbohydrate needs. The second phase 
will consist of definitive studies of the effect of imposed exercise on the 
systemic effects of tumors. 

Progress: The feasible limits for dietary and exercise regimens have been 
established. The effect of exercise on response to tumor has been studied 
for W256 carcinosarcoma. The interaction of exercise and dietary carbohydrate 
source in presence of W256 is now being studied. 



Significance to Cancer Research: Information concerning the effects of 
imposed or encouraged exercise and of predominance of simple or complex 
carbohydrates in diet, or retardation of wasting and hypophagic effects of 
cancer will be of great importance to knowledge of origins and possible 
methods of combating cachetic decay in cancer. 

Project Officer: Dr. Seoras D. Morrison 
Program: Biology Support 
Technical Review Group: Ad Hoc Committee 
FY 81 Funds: $42,223 



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