BIOLOGY AND DIAGNOSIS AREA

Program Coordinators: Dr. William D. Terry, U.S.A.
Dr. Yuichi Yamamura, Japan
Principal Advisors: U.S.A.:
Dr. Ira Pastan
Dr. Richard Hodes
Dr. Myron Melamed

Japan:
Dr. Yoji Ikawa
Dr. Toshiyuki Hamaoka
Dr. Yoshio Tenjin

ADMINISTRATIVE REPORT

A. SEMINARS

1) Cellular & Molecular Biology Area

Src and Leuk Genes
Location: National Institutes of Health, Bethesda, MD, USA
Dates: November 12-14, 1979
Participants: 14-U.S.
8-Japanese
Summary:
The meeting lasted for 2-1/2 days with a dinner at the end of the first day. There was an equal division of time between formal presentations and informal discussions. A lot of new information was presented and there was an opportunity to discuss in detail some previous work in which there was some disagreement among the investigators present.
The initial talk was by Dr. Karen Beemon who presented evidence that there was specific phosphorylation of tyrosines in Rous sarcoma virus transformed cells. This phosphorylation appears to be catalyzed by the transforming gene product of Rous sarcoma virus. Phosphotyrosines in proteins are novel and of probably great regulatory significance. Next, Dr. Jim Neil discussed recovery of sarcoma proteins from recovered avian sarcoma viruses. Following his presentation, Dr. Hanafusa also discussed similar experiments. It is clear that Rous sarcoma virus can recombine with host sequences to generate new kinds of transforming viruses. This exciting finding is consistent with the notion that a protein very similar to the sarcoma protein is present in normal chicken cells. Dr. Tadashi Yamamoto discussed recent experiments he has carried out on unusual cDNA clones isolated from avian sarcoma virus transformed cells. Dr. Yamamoto presented evidence that there are additional sequences present at the end of certain messenger RNAs present in ASV transformed cells that were previously unexpected. One of the clones he has isolated appears to have a promoter which probably is the site of initiation of Rous sarcoma virus transcription. Dr. Yamamoto also proposed a new model by which sarcoma-defective varients are generated. Dr. Sadaaki Kawai then discussed a new mutant of Rous sarcoma virus defective in transformation derived from a classical mutant ts-NY68. The afternoon session began with an exciting presentation by Dr. Ray Erikson describing the phosphorylation catalyzed by the Rous sarcoma virus transforming gene product that he has identified. Then Dr. Gilbert Jay described experiments in which phosphorylation of proteins was observed in cell free extract of RSV transformed cells.
Dr. Asim Dasgupta from Dr. Baltimore’s laboratory described the characterization of a phosphoprotein from cells transformed by the Abelson strain of murine leukemia virus. This phosphoprotein also contains a phosphotyrosine.
Dr. Oda reviewed work previously done in his laboratory on the phosphorylation of proteins in virally transformed cells. Dr. Yoshikura then described experiments in which there is cocarcinogenesis produce by both viral transformation and chemical transformation.
The morning of the second day was devoted initially to studies by Drs. Shih and Scolnick on the characterization of the transforming protein of Harvey sarcoma virus. This protein is a membrane protein and has the capacity to bind GTP. Dr. Tsuchida also described studies on a murine sarcoma virus; he presented evidence on the linear map of the double stranded DNA present in Kirsten sarcoma virus transformed cells. Dr. Yahara described experiments in which he used antibodies to tubulin to examine various leukemic cells that had alterations in the cytoskeleton. He hopes these studies will be of clinical usefulness. Dr. Mark Willingham described recent studies on the ultra-structural localization of src proteins in cells transformed both by Rous sarcoma virus or by Kirsten sarcoma virus.
In the afternoon session, Dr. Yoshida began the discussion with identification of SFFV-specific sequences coding for a SFFV-specific glycoprotein. Then Dr. Leonard Evans discussed the ribonucleic acids of the defective and nondefective component of Friend anemia and polycythemia viruses. There are prototypes of a new class of retroviral transforming genes. Dr. Ikawa then discussed experiments in his own laboratory on the characterization of the SFFV-specific glycoprotein, gp-55. Dr. Toyoshima described some properties of a newly isolated avian sarcoma virus. Dr. Michael Lai discussed experiments on leukemia specific sequences in avian erythroblastosis virus. The third day of the meeting was devoted to a special talk by Dr. Wallace Rowe in which he reviewed the current status of the mink cell focus forming virus. This is a very unusual transformation system that may have relevance to human cancer. Finally, Dr. Paul Neiman discussed experiments characterizing malignant lymphomas in chickens induced by avian sarcoma virus. He looked at the organization of the viral sequences in the tumor as the tumor progressed from a small to a large tumor. The rest of this meeting was devoted to general discussion of the meeting and to plans for a possible meeting in the following year.


2) Immunology Area: Biology, Genetics, & Preclinical Immunotherapy
Mechanisms of Host Tumor Immunity and theoretical Basis for Tumor Immunotherapy
Location: Sheraton Maui Hotel, Kaanapali Beach, Maui, Hawaii
Dates: October 8-10, 1979
Participants: 9-U.S.
7-Japanese
Summary:
The first joint seminar of this Immunology Program Area involved presentation and discussion of research in the areas of (1) antigenic nature of tumor cells, (2) fundamental immunologic processes in host responses to tumor, and (3) new approaches and preclinical trials in tumor immunotherapy.
The antigenicity of tumor cells in experimental systems was critically evaluated in presentations which stressed the importance of rigorous identification of tumor-associated antigens. In addition, the relevance of such antigens to tumor sensitivity in immunotherapy was evaluated both in formal presentation and subsequent discussion. Extensive experimentation was reported by both U.S. and Japanese participants in studies of the fundamental immunologic mechanisms of anti-tumor responses. The control of immune responses by specific immuno-regulatory processes was described in a number of tumor systems and new information concerning effector mechanisms mediated by macrophages and natural killer cells was presented. In the final sessions of this meeting, new approaches to tumor immunotherapy were discussed, based upon current experimental findings in areas of basic immunology and tumor immunology. The participating scientists from both Japan and the United States were among the most talented and productive of investigators working in basic immunology, immunogenetics, and tumor biology. This unique opportunity for interaction among these scientists generated not only a valuable exchange of scientific information, but a critical forum for the focussing of basic science perspectives on the development of new approaches to tumor therapy.


3) Diagnosis Area: Automated Cytology & Development of New Techniques
Cytology Automation
Location: Sheraton Universal Hotel, Los Angeles, CA, USA
Dates: January 27-29, 1980
Participants: 12-U.S.
8-Japanese
Summary:
The participants assembled informally at a reception in the Hotel on Saturday evening, January 26. The formal meetings began on Sunday morning, January 27 An effort was made to represent reports on instrumentation and data analysis techniques the first day, cancer detection systems the second day, and an overview of hematology (leukemia) and other applications the third day. It was the consensus of the participants that the formal presentations and the discussions that followed had covered the important present applications of automated cytology.
Since the last meeting, there has been progress in the development of sample preparation techniques (Drs. Uei, Kishigami, Wied, Tanaka, and Mendelsohn), in the development and application of automated slide-scanning instrument systems for cervix cancer detection (Drs. Tanaka and Wied), in a promising flow cytometry system for detection of cervix cancer and precancerous lesion (Dr. Tenjin) and in a flow cytometry system for detection of urinary bladder cancer (Dr. Melamed). New slide scanning image analysis techniques were developed for detecting malignant cells within cell clumps based on multispectral measurements (Dr. Noguchi), and for recognizing adenocarcinoma cells in cervico-vaginal smears based on nuclear overlap and inter-nuclear distance as well as more conventional nuclear shape and staining features (Dr. Onoe). A new image sensing device was described for rapid high resolution scanning based on a charge coupled device in linear array composed of 2048 sensor elements (Dr. Takahashi). The role of flow cytomtry in monitoring effects of steroid hormones on DNA RNA and cell cycle of hormone-sensitive cell lines was described (Dr. Nishiya). The possibility of increasing resolution of cell structure in flow cytometry was discussed following a description of 3-dimensional slit-scanning in flow and tv imaging of cells in flow (Dr. Wheeless). Cell classification by means of cytochemical markers in leukocytes and leukemic cells was reviewed, primarily as applied in flow cytometry (Dr. Ornstein). A new flow cytometric technique was described for sub-classifying acute lymphoblastic and non-lymphoblastic leukemias (and the chronic leukemias) by RNA index, and identifying aneuploid stem-lines by precise DNA measurements (Dr. Andreeff). The present clinical usefulness of this flow cytometry analysis of the leukemias was emphasized, not only for classification but also in following cell kill kinetics, identifying relapse at an early stage and identifying biphasic phenotypic hematopoietic neoplasms that appear to be more common than hitherto believed and may require treatment strategies. The technology involved in producing monoclonal human antibodies was reviewed, with emphasis on the antibodies to sub-classes of normal leukocytes which will soon be commercially available and may influence the present classification scheme for leukocytes (Dr. Goldstein). Preliminary experiments with leukemic cells indicate a strong possibility for developing type-specific monoclonal antibodies for classification of leukemias. There was also discussion about the possibility of producing antibodies to carcinoma cells. Data analysis similarities and differences in high resolution scanning systems and flow cytometry systems were described (Dr. Sharpless). The need for computer analysis of large numbers of point measurements in the image of a cell was compared with the analysis of large numbers of cells measured from a specimen analyzed by flow cytometry. The reported results of clinical trials of cervix cancer detection with many different high resolution scanning systems over the last 25 years was reviewed, and steady progress was noted (Dr. Kamentsky). Flow cytometry trials for cervix cancer detection was more recent, and have not yet shown significant improvement. The cost and performance requirements for a successful cervix cancer detection instrument was calculated, based on various receiver operating curves and reasonable cost estimates of manual and machine methods (Dr. Castleman). It was pointed out that a successful instrument will very likely have to operate far out on the curve, yielding very few false positive results at the expense of many false negative measurements-a conclusion that elicited considerable discussion and surprise. Methods for statistical comparison of machine and human performance were also discussed (Dr. Prewitt). Finally, the broad ranging applications of flow cytometry at Los Alamos (Dr. Mullaney) and at Lawrence Livermore Laboratories (Dr. Mendelsohn) were described, and recent studies at the Battelle Institute using single cell measurements to establish thresholds for distinguishing specific cell types by flow cytometry (Dr. Mullaney). These presentations included a review of the rapid recent progress in automated karyotyping by flow cytometry, analysis of sperm DNA and form changes as a result of toxic chemical or mutagen effect, methods for measuring mutagen effect on other cell types, and new developments in multilaser, multiparameter flow cytometry. The U.S.-Japan meeting on cytology continue to be extremely productive for participants from both countries, particularly in guiding the direction of research and development as new observations and technology are presented. Therefore it is recommended that the meetings be continued at approximately 12-18 month intervals. It was recommended that the contents of the next meeting, to be held sometime between April 1, 1981, through September 30, 1981, be approximately 50% on cancer detection, diagnosis and monitoring, and approximately 50% on new cancer-related applications of flow cytometry in immunology, cell biology, etc.



B. EXCHANGE OF SCIENTISTS
1) Kiyoshi Takatsu, Assistant Professor, Department of Oncogenesis, Institute for Cancer Research, Osaka University Medical School, was dispatched to the Scripps Clinic and Research Foundation, for one month from January 7, 1980, and studied the immunobiological property of tumor-associated transplantation antigens in the Department of Molecular Immunology under the supervision of Prof. Ralph A. Reisfeld. Besides, he visited Dr. Martin Dorf in Department of Pathology, Harvard Medical School and Dr. Christopher Henney in Fred Hutchinson Cancer Center to exchange the informations of recent progress in cancer immunology.

2) Dr. Masuo Obinata, Senior Investigator, Laboratory of Viral Oncology, Cancer Institute Tokyo, was dispatched to the Clinical Hematology Branch, National Heart, Blood and Lung Institute in N.I.H., for one month from February 10-March 15, 1980, and studied the switching mechanism of globulin gene expression in mouse fetal development under the supervision of Chief of the Branch, Dr. Arthur W. Nienhuis. On a way of his visiting to the N.I.H., he attended the IVth Annual Symposium on molecular basis of cell-cell interaction at San Diego. During his stay in U.S.A., he also visited Dr. Shigeru Sassa in Rockefeller Univ., Dr. William E. Hahn in Univ. of Colorado, Medical Center, and Dr. Brian J. McCarthy in University of California at San Francisco, to exchange the information of mechanism of gene expression.

3) Dr. Yoshio Noguchi, Senior Researcher, Information Sciences Division Electrotechnical Laboratory, Tokyo, spent 3 months from November 11, 1979 to February 8, 1980 in National Institute of Health to study the hybrid-system in automated cytology, under the supervision of Dr. Judith Prewitt. During his stay in U.S.A., he visited Dr. Leon L. Wheeless, Stron Memorial Hospital, Univ. of Rochester, Dr. Myron R. Melamed, Memorial Sloan-Kettering Cancer Center and Dr. Ron Jensen, Lawrence Livermore Laboratory to investigate flow-through instrument.

C. EXCHANGE OF MATERIALS

Dr. Ichiro Azuma sent the following immuno adjuvants:
2 x 50 mg quinonyl-N-acetylmuramyldipeptide
5 x 100 mg-heat-killed cells of Propionibacterium acnes
5 x 100 mg-cell-wall skeleton of P. acnes
These samples were distributed to Dr. Ronald Herberman (National Cancer Institute) and Dr. Carl Nathan (Rockefeller University) to examine adjuvant activities on natural killer cells and macrophages. By these examinations, the correlation between the activities for induction of natural killer cells, tumoricidal macrophages and interferon will be delineated.