REPORTS ON SEMINARS

(1) Seminar on "Immunology"

The conference on "Immunogenetic Analyses of the Expression of Tumor Antigens and Responses to Tumors" was organized by Dr. T. Hamaoka from Japan and Dr. Richard J. Hodes from the United States. There were 24 formal presentations by 14 Japanese and 5 U.S. participants, each followed by informal discussion. Emphasis was placed on the genetic and immunologic study of tumor antigen expression, the basic cellular and humoral mechanisms of host immune response, and the development of preclinical models of immunotherapy.
A series of presentations described the existence and identification of tumor-specific antigens, the relationship of these antigens to oncogenes, and the molecular biology of oncogenesis. Dr. Masaru Taniguchi described the existence of melanoma-specific antigens widely shared among various animal species. These studies were conducted employing both monoclonal hybridoma antibodies and cytotoxic T lymphocytes. Both serologic and cellular reagents identified the existence of cross-reactive antigens present on melanoma cells of the mouse and human, but absent in a series of control normal and malignant tissues. The finding of such species cross-reactive tumor antigens was also applied to immunotherapy studies employing radio-labeled monoclonal anti-melanoma antibodies, which were successful in preliminary immunotherapeutic trials in the mouse. Dr. Ko Okumura described the existence of human embryonic tumor antigens which were cross-reactive with the murine F9 antigens, as determined by studies of sera from patients with germ cell-related malignant diseases. The application of such findings to both diagnostic and therapeutic purposes was discussed. Dr. Noboru Kuzumaki studied the relationship between the src gene expression and expression of tumor-specific antigens. Employing a series of RSV mutants demonstrated that expression of the RSV src gene correlates with tumor-specific antigen expression. Immunologic studies of the human adult T cell leukemia (ATL) were presented by Dr. Kazuo Sugamura. Evidence was presented that strongly suggests a causal relationship between the ATL virus and the human adult T cell leukemia seen in high incidence in southern Japan. In vitro transformation experiments have substantiated the transforming potency of virus associated with the cells of human ATL patients. The association of the ATL antigen in both leukemia patients and apparently normal individuals in related geographic areas was also described. The molecular biology of the human ATL retrovirus was discussed by Dr. Mitsuaki Yoshida. The viral genome was studied by restriction enzyme analysis. In particular, an analysis of integration sites showed differences between integration sites in a patient with T cell leukemia and the integration sites in in vitro infected cultured B cell lines. The data suggested random integration with a clonal origin of leukemia from infected cells. Dr. Stanley Korsmeyer described genetic rearrangements within human lymphoid neoplasms. Rearrangement of immunoglobulin genes in human lymphomas and leukemias was studied. Such analysis was utilized to interpret lymphoid malignancies in terms of normal differentiation sequences of the B cell and T cell lineages.
Several new technologies for future application to tumor immunology were introduced. Dr. Tsuyoshi Uchida described new techniques for gene transfer that allow the transfer of unusually large segments of genetic material and other macromolecules. Dr. Tadamitsu Kishimoto described studies employing microinjection for analysis of cell differentiation and activation mechanisms. These studies allowed the introduction of cytoplasmic factors from activated B cell lines which were then capable of inducing the differentiation of IgG secretion in resting B cell lines. Takeshi Watanabe presented techniques of nuclear transplantation and microinjection that allow the study of control mechanisms for cell surface receptor expression. Using such techniques, the relative contributions of nuclear and cytoplasmic factors on B cell responses to mitogenic stimulation were analyzed.
Another presentation covered the area of host responses to tumors. Dr. Richard Hodes described two pathways of suppressor T cell control of B cell activation. These pathways, involving distinct T cell subpopulations, were characterized by both antigen specificity and I-A subregion genetic restriction in their activation and effector functions. Dr. Mark Greene presented experimental data in a system that allowed the cytotoxic T cell killing of reovirus-infected target cells or of anti-idiotypic hybridomas with specificity for the antireovirus 3 idiotype. MHC-restricted kill of anti-idiotypic hybridomas was demonstrated, and was shown to be inhibited by either antireovirus 3 antibody or by anti-MHC antibody. These findings suggested the existence of two discrete receptors on cytotoxic T lymphocytes. Dr. Richard Hodes then presented information analyzing the genetic restriction of B cell-accessory cell interactions. These findings demonstrated that, in the T cell-independent response to TNP-Ficoll, there was an MHC-restricted interaction between B cells and lymphocytes. In addition, it was demonstrated that MHC-restricted B cell recognition was subject to adaptive differentiation during B cell development. These findings suggested that B lymphocytes, as well as T lymphocytes, are capable of both MHC-restricted recognition and adaptive differentiation. Dr. Steven Burakoff characterized the cell interactions regulating cytotoxic T lymphocyte responses to infectious and oncogenic viruses. The technique of "exon shuffling" from cloned L^d and D^d genes was described. Using L cell transfection, and the study of transfected cells as CTL targets, it was possible to analyze the importance of discrete exon gene products in the determination of both alloantigens and restriction elements for T cell recognition. Dr. Shigeyoshi Fujimoto studied functional killer T cell receptors in a system of A/J antisyngeneic tumor S1509a. He was successful in generating a CTL clone to tumor antigens, and then proceeded to generate monoclonal hybridoma antibodies to this cytotoxic T cell line in the syngeneic A/J mouse. The behavior of this monoclonal antibody suggests that it is specific for the antitumor T cell receptor. The class of human lymphocyte functional antigens (LFA) associated with T lymphocyte-mediated cytolysis was discussed by Dr. Steven Burakoff. Using a panel of monoclonal antibodies, he outlined a series of LFAS which appeared to be relevant to the cytotoxic effector function of human T cells, either by their action upon cytotoxic T lymphocytes or by their action upon target cells. Dr. Masaji Okada analyzed a series of human monoclonal lymphokines through the study of T cell hybridoma products. These factors included IL-2, killer helper factor (KHF), B cell growth factor (BCGF), and B cell differentiation factor (BCDP). The structure and expression of genes for interferons was discussed by Dr. Tadatsugu Taniguchi.
Dr. Susumu Tonegawa presented a review and analysis of the mechanisms for generation of diversity in antibody genes. Recombinatorial mechanisms, junctional insertions, and somatic mutations, and the mechanisms by which these generate ultimate antibody receptor diversity, were discussed. Dr. Stanley Korsmeyer analyzed related issues in the description of a hierarchy of immunoglobulin gene recombination in both normal and human lymphocytes. Through this hierarchy, heavy chain and light chain rearrangements appeared to occur in an ordered and regulated sequence.
This seminar also addressed the subject of preclinical trials for tumor immunotherapy. Dr. Hiromi Fujiwara described the genetic control of hapten-reactive T helper cell responses. TNP- and sulfhydryl-reactive AEDANS-specific T helper cells were studied in the generation of cytotoxic T lymphocyte responses to modified self. It was demonstrated that these two anti-hapten self-responses were under independent Ir gene control, and that helper cells specific for each of these haptens were capable of enhancing the tumor-specific CTL response in vitro. Dr. Toshiyuki Hamaoka described a series of studies based on the same principles, which allowed application to tumor immunotherapy models in vivo. A model system was described which involved the elimination of suppressor cells by pretreatment of animals and the subsequent priming to hapten, followed by immunization with hapten-coupled tumor. It was demonstrated that such a protocol allowed the induction of potent tumor immunity. Moreover, when applied as an immunotherapy protocol, an optimalized regimen allowed substantial immunotherapeutic responses in the mouse model. Dr. Yoshiyuki Hashimoto has studied a model of tumor immunotherapy employing monoclonal antibody incorporated in liposomes. Liposomes were constructed incorporating the tumor chemotherapeutic agent actinomycin D, to which antitumor monoclonal antibodies had been conjugated. It was found that these targeted liposomes were effective in an in vivo tumor immunotherapeutic model. Data suggesting the tumor specificity of these effects were also presented. Mark Greene described the use of tissue-specific monoclonal antibodies in tumor diagnosis. Radiolabeled tissue-specific antibodies were shown to be capable of selectively binding to human tumors of specific tissue origin, and to allow diagnostic imaging of human tumor implants in nude mice.
The final session of this meeting included a general discussion of the basic tumor biology and immunology presented, and most particularly, of the application of these principles to future studies of tumor diagnosis and immunotherapy. It was noted that many of the cellular immunologic, immunogenetic, and molecular biologic findings of the past years have now allowed the generation of rational approaches to tumor diagnosis, prevention, and therapy. It is anticipated that a new generation of models for tumor immunotherapy and diagnosis will evolve based upon newly established findings.

(2) Seminar on "Cellular and Molecular Biology"

The fourth meeting of the Cellular and Molecular Biology sub-area of the U.S.-Japan Cooperative Cancer Research Program was convened at the East-West Center, University of Hawaii, on January 17, 1983. This meeting, entitled "Gene Transfer in Animal Cells and Activation of Cellular Oncogenes," was organized by Dr. Yoshio Okada and Dr. George F. Vande Woude. There were seven participants from Japan and six from the United States.
The subjects of the first session centered on cellular and viral oncogenes. Dr. Channing Der reviewed the dominant transforming genes that have been studied at the Dana Farber Cancer Center. These studies have been at the forefront of the cellular onc gene field; specifically, they have shown that the viral onc gene ras^H and ras^K family are the dominant transforming genes transfered to NIH 3T3 cells in certain human bladder (ras^H) and human lung and colon (ras^K) tumor cell lines. Dr. Der also stated that the non-myc beta lymph gene from ALV-induced chicken bursal lymphoma was related to the dominant gene transferred from human Burkitt lymphoma cell lines.
Dr. Michael Wigler from the Cold Spring Harbor Laboratory described the isolation and characterization of three human dominant transforming genes that are related to the ras gene family. Two were the same genes that Dr. Der had described, i.e., the ras^H in a bladder carcinoma cell line, and the ras^K in the human lung carcinoma line. He described the single-base change in the human bladder ras^H gene that results in a codon change from a gly to val in the gene product, and causes cellular transformation in the NIH 3T3 cell assay. The ras^K gene of the lung carcinoma has been partially cloned and may encompass 40kb of genomic DNA. The third ras-related gene Dr. Wigler described was cloned from a human neuroblastoma cell line. The onc gene has sequence homology with both the ras^H and ras^K genes, but is unique in structure and therefore represents a third member of the ras family. It is not known how these later two transforming ras members differ from their normal homologues.
Dr. Hiroshi Amanuma of the Cancer Institute described the sequence of the gp55 of spleen focus-forming virus (SFPV). This protein is a fusion product consisting of an N-terminal portion derived from an MCF recombinant MULV virus, pl5E of MuLV, and a C-term amino acid sequence that is specific for SFFV. Specific constructs for identifying the important leukemogenic structural features of this gene are currently being developed.
Dr. Barthol Sefton of the Salk Institute reviewed the extensive characterization that he and Tony Hunter have made of the tyrosine protein kinases encoded by the viral onc genes. The kinases of the src, yes, fps/fes and abl gene products phosphorylate some of the same cellular substrates. Most recently, these investigators have shown that the src, abl, and ras gene products all contain lipid, suggesting a mechanism for directing these proteins to the plasma membrane.
Dr. Eiichi Soeda of the National Institute of Genetics described transfection studies into 3Y1 rat and NIH 3T3 cells with a plasmid containing only the origin of polyoma virus and lacking functional tumor antigens. This transfected DNA appears to induce foci in recipient cells by some unknown mechanism. The transformed-transfected cells are positive for polyoma sequences by Southern analysis.
Dr. Yogi Ikawa from the Institute of Physical and Chemical Research described the construction of a retrovirus vector. They have placed the chick lens delta crystalline gene into this vector and after transfection into NIH 3T3 cells have detected expression. They have also tested the viral mos onc gene in this vector.
Dr. William Hayward of the Sloan-Kettering Institute for Cancer Research reviewed the promoter insertion model for leukemogenesis. He described in situ hybridization-mapping experiments that localized the human c-mos and c-myc loci on chromosome 8 at positions q22 and q24, respectively. Dr. Hayward pointed out that these locations correspond to breakpoints in translocations observed in acute non-lymphoblastic leukemia (q22) and in non-Hodgkin's Burkitt's lymphomas (q24).
Dr. T. Taniguchi of the Cancer Institute gave a comprehensive presentation on the expression of human interferon (IFN) genes introduced into cultured mouse cells. He has identified a region upstream from the alpha and beta IPN genes that is conserved (in mouse IFN-beta as well), and may be associated with a promoter-regulator region of the gene. Thus, the human IFN-beta₁ gene was shown to be inducible in mouse cells. Dr. Taniguchi studied the effects of deletions in this region on expression IFN B1. A -100 to -90 base-pair inverted repeat appears to be essential for induction of this gene in response to poly I.poly C.
Dr. Moses Chao of Columbia University described experiments with mouse-human hybrid beta globin genes in mouse erythroleukemia cells (MEL). These studies were initiated to identify the region(s) responsible for globin mRNA expression during erythroid differentiation. Dr. Chao showed that sequences 5' to the beta globin structural gene up to-78 base-pairs can be removed without affecting the loss of inducibility of the beta globin mRNA in the MEL cells.
Dr. Yoshio Okada of Osaka University, a coorganizer of this meeting, described procedures that have been developed in his laboratory for introducing macromolecules into living cells. Dr. Okada is a pioneer in developing these protocols; his most recent modification, RBC hemolysis and ghost trapping of macromolecules followed by the Sendai virus fusion, appears to be an efficient way for quantitative introduction of molecules into cells. This system works efficiently for small proteins like BSA, but the RBC ghosts fail to efficiently trap larger proteins or DNA.
Dr. Masahiro Ishiura of the National Institute for Basic Biology described several parameters that yielded efficient phage particle-tk gene transfer to mouse Ltk^- cells. He was able to obtain efficiencies of one transformant/10⁵ phage/10⁶ cells. By this protocol, he was able to transfect the human tk gene from a lambdaphage gene library (one mouse Ltk⁺ transformant/2.8 x 10¹⁰ PFU), and is in the process of cloning this gene from the mouse Ltk⁺ transformant.
Dr. Takeshi Seno of the Saitama Cancer Center Research Institute described studies with a mouse thymidylate synthase-negative cell mutant that was transformed with the human enzyme by transfection. It was possible to study the physical properties of the human enzyme in these cells and to determine the levels of expression from the transfected human gene. Higher levels of the human thymidyiate synthase appear to be required to support norma1 growth, but the presence of the mouse enzyme suppresses the production of the human thymidylate synthase. His studies indicate that the higher production of the latter is not due simply to the gene copy number.
The meeting was well received by ail the participants and, as previously determined, the appropriate atmosphere, together with the important constraint on the size of the meeting, provided an extraordinary opportunity for extensive and meaningful discussions. DNA transfer and onc genes were timely subjects that served to maintain a keen interest and enthusiastic pace.

(3) Seminar on "Cytology Automation"

The seminar on "Cytology Automation" was held February 19-22, at the Memorial Sloan-Kettering Cancer Center, New York, New York. On Saturday, February 19, 1983, the evening prior to the opening scientific session, Drs. Y. Tenjin and M. Melamed met to review the present status of the U.S.-Japan Cooperative Program on Cytology Automation. Prom both the Japanese and the U.S. perspective, it was clear that the emphasis on instrument development over the past 5 years has been a wise decision. Research flow cytometers and high resolution scanning microscopes now are capable of carrying out very precise measurements of single cells and cell populations. For the next decade, it is felt, instrument development should continue at a lower level of effort, with much more emphasis placed on biologic research. In particular, there is a need to develop biologic methods and new probes for cell analysis that would lead to more effective clinical, as well as research, applications of the new instruments. Clinical applications, for example, should be expanded beyond cancer detection, the initial goal, to include monitoring in vivo cancer treatment effects; developing new in vitro assays of cancer cell cycle kinetics for evaluating chemotherapeutic agents; subclassifying cancer cells with fluorescence-tagged monoclonal antibodies; using chromatin structural differences to recognize or subclassify cancer cells; and, perhaps, using fluorescence-tagged complementary nucleic acid probes to identify intracellular viruses and gene products.
The formal scientific presentations were held in a conference room of the Memorial Sloan-Kettering Cancer Center, February 20-22, 1983. On the morning of the first day, new methods of specimen preparation and tumor cell enrichment were presented. Drs. Walle and Kodama described a continuous Percoll density gradient for sub-fractionating cell suspensions that was highly effective in concentrating lung cancer cells. Dr. M. Takahashi presented the results of cell sorting by flow cytometry for enriching cancer cells from uterine cervical specimens. Dr. N. Tanaka described a device that automatically syringed cell specimens to obtain a single cell suspension and then prepared a monolayer smear for high resolution scanning.
Several important advances in instrumentation were presented. Drs. M. Takahashi, N. Yoshimi, and H. Aoki used an array of charge coupled diodes (CCD) to rapidly scan a slide area of 15 square micrometers at 1.0 and 0.25 micrometer intervals, with quite good resolution. Dr. J. Steinkamp described a new three-laser, three-station system for multiparameter flow cytometry using argon and krypton lasers. He was able to measure simultaneously DNA, RNA, and protein content of CHO cells that were stained by Hoechst 33342, Pyronin Y, and fluorescein isothiocyanate. An ultrafast laser scanner microscope was reported by Dr. P. Bartels; with this instrument, scanning rates of 64 megahertz were obtained at two wavelengths. Thus, a 2x2 cm slide could be scanned in approximately 60 seconds, with excellent cell image detail. A multiprocessor, microprocessor system is being designed to process the enormous data stream in real time.
The results of automated screening for uterine cancer using CYBEST Model 4 were reported by N. Tanaka. While similar to Model 3, it is much smaller and requires only 3 minutes per specimen. Dr. Wheeless reported results of a single blind study of gynecological cytology specimens examined by multidimensional slit-scan flow cytometry. Of 740 specimens, including 156 that were abnormal, 17.6% were false positives, but only 2.8% were false negatives. Dr. D. Rosenthal reported on the current status of the Jet Propulsion Laboratory-UCLA project to develop a high-resolution cytology scanning instrument. Algorithms have been developed based on features extracted from 10,000 normal and abnormal human uterine cervical cells, and instrument design specifications have been determined.
Other clinical applications of high-resolution and flow cytometry were presented in the afternoon of the first day: Drs. Tenjin, Sugita, Muroya, and Sugishita reported studies of the flow cytometry of cycling uterine cervical cancer cells. Dr. Bartels pointed out the difficulties inherent in distinguishing rare cancer cells in a clinical cytology specimen from "false alarms" due to artefacts. The reported discovery of measurable changes within many normal-appearing intermediate cells that accompany cancer provides a novel opportunity to circumvent this problem. He estimates that samples of only 400-800 intermediate cells could provide correct prescreening diagnoses, and that this marker expression ranges from 90% to 70% of cases. Drs. Wied and Bibbo reported that computing two indices from Feulgen-stained smears (percent aneuploid cells exceeding 5N DNA and deviation of measured DNA from a lymphocyte standard) would provide a clinically useful "malignancy index."
The remaining afternoon session dealt with applications of flow cytometry in hematology. Drs. S. Takamoto, R. Veda, K. Ota, and T. Takahashi reported on the specificity of three monoclonal IgM antibodies obtained by immunizing with HL-60 cells. Using flow cytometry and rosette analysis, they were able to demonstrate good specificity for myelocytes and monocytes in normal and leukemic patients. Drs. K. Nomura and T. Watanabe reported a significant imbalance of T cell subsets in patients with gastric cancer, measured by flow cytometry with fluorescence-labeled monoclonal antibodies, and related this to differences in prognosis at each stage of disease. Dr. M. Andreeff reported on 430 acute leukemias and 100 non-Hodgkin's lymphomas studied by flow cytometry DNA/RNA analysis over a 6-year period. His data showed useful prognostic information in DNA content, RNA content, proliferative index, chromatin structure, and rate of cell kill with therapy. Dr. A. Redner examined cerebrospinal fluid specimens from 23 patients with acute leukemia; 14 had documented CNS relapse, and in 6 there were aneuploid cells identified by flow cytometry. In three of the six cases, leukemia could not be identified in the spinal fluid by conventional cytology.
Biologic and medical applications of automated cytology were discussed in additional presentations on the second day of the meeting, February 21. The morning session began with a description of the CCD image analysis system applied to the grading of bronchial epithelial atypia in histologic sections. Drs. M. Takahashi, N. Yoshimi, and H. Aoki used this system to demonstrate differences in distribution of nuclear area and integrated nuclear density of normal, atypical, and metaplastic bronchial epithelium. Drs. L. Koss and A. Sherman then described results of bladder cancer diagnosis by high-resolution image analysis of exfoliated epithelium in the urine. They used a hierarchical triage to exclude degenerating cells that were difficult to analyze, and successfully diagnosed 15 of 15 bladder cancers. Bladder cancer diagnosis by flow cytometry of bladder irrigation specimens was presented by Dr. L. Staiano-Coico. She reported 95% accuracy in detecting carcinoma in situ, based on a finding of aneuploid cells, and was able to distinguish low- and high-grade papillary tumors. Flow cytometry detection and monitoring of bladder cancer is now routinely applied to patients on the Urologic Service of Memorial Hospital in New York City, and appears ready for implementation at other hospitals. Dr. J. Steinkamp reported studies of bronchial lavage specimens analyzed by flow cytometry, and proposed using this method to assess effects of environmental air pollutants. In the last presentation of the session, Dr. L. Koss described striking differences in prognosis of human colon carcinoma, according to DNA stemline as measured by flow cytometry.
The second session of the morning addressed several applications of automated cytology to cell biology. Dr. M. Takahashi described a simple and convenient two-dimensional table from which to read the c.v. of DNA distribution from the channel number of the G₁ Peak and the "bluntness index" (defined as the ratio of mean height of the two adjacent channels to the height of the peak). Dr. Z. Darzynkiewicz described a new flow-cytometric technique for detailed cell cycle kinetics analysis based on simultaneous measurement of DNA content and chromatin structure of the cells under study. Using this technique, drug effects on the cell cycle and drug-induced changes in nuclear chromatin can be assayed with precision. Dr. M. Takahashi proposed a new computer program for analysis of DNA histograms obtained by flow cytometry. Using this, he reported the S-phase distribution of three cell lines studied to be relatively flat, rather than parabolic, with a sharp bend upward near G₁ and G₂M. Drs. S. Takamoto and K. Ota studied the effect of the vital DNA stain Hoechst 33342 on a T cell leukemia line, and found that it caused transient slowing in cell cycle progression and cloning efficiency. Dr. C. Cordon-Cardo described the present status of tumor-associated monoclonal antibodies as new markers for identifying and subclassifying tumors in histologic sections, cytology smears and by flow cytometry. The most promising thus far are those developed for urinary tract tumors.
The afternoon of the second day was devoted primarily to studies of drug effects in clinical situations. However, in the first presentation, Dr. P. Higgins reviewed the new technique of in situ hybridization of nucleic acids, and discussed the possibility of tagging complementary nucleic acid probes with a fluorescent dye so they might be used for automated cytology. White this is technically feasible, it probably will be used only for high copy sequences such as viruses. Dr. F. Traganos described the use of flow cytometry to measure drug effects on the cell cycle, and showed cell cycle changes induced by several different drugs. Drs. Y. Tenjin, M. Sugita, T. Muroya, and T. Sugishita reported a study of combined chemotherapy (cisplatin, endoxan, adriamycin, and 5-FU) used in four cases of peritoneal carcinomatosis with ascites, and were unable to relate DNA measurements or cytologic morphology to clinical response.
The final session of the afternoon began with a report from Dr. K. Nomura, in which he assessed the effect of ACNU and Mitomycin C on the cell cycle kinetics of four glioma cell lines. He found that an accumulation of cells in SG₂M predicted sensitivity and cell kill by the drug, and this was confirmed in multi-cell spheroid cultures and in Fisher rats with brain tumors. Dr. I. Nishiya used flow cytometry to study site-directed chemotherapy, and described the effect of a nitrogen mustard-estradiol conjugate on the cell cycle kinetics of a cultured endometrial adenocarcinoma. The last presentation of the afternoon was a report by Dr. D. Evenson, who described a new method of assaying drug effects on spermatogenesis by means of flow cytometry. The assays may be conducted on semen or on testicular cells, and they indicate abnormalities of sperm maturation as well as sperm number, viability, and motility. The assay is highly sensitive to drugs at low doses and to low levels of irradiation. It has potential applicability in monitoring environmental chemical pollutants and radiation exposure.
Tuesday morning, February 22, the third and final day of the meeting, there was a presentation by Dr. C. Herman, who described machine-measurable changes in normal-appearing intermediate squamous cells of patients with associated uterine cervical neoplasia. These changes were similar to those described earlier by Dr. Bartels and involved nuclear and chromatin structure.
Dr. Koss then chaired a discussion of the potential clinical applications of automated cytology. The most immediate and clinically most important applications at present and in the near future are likely to be in hematology. These will include subsetting normal leukocytes by surface antigens using monoclonal immunofluorescent reagents; in leukemia, applications will include classification by DNA/RNA with surface antigens, and monitoring treatment regimens by cell-kill kinetics. Cancer detection using flow cytometry is feasible at present for carcinoma of the bladder. Further work remains before uterine cervical cancer detection will be practical, but it appears feasible in principle. One of the major applications, and certainly one of the greatest needs, will be in assaying solid tumors by automated cytology of needle aspiration specimens. Cell cycle kinetics can be analyzed with extraordinary detail and high precision using flow cytometry, and has been used primarily to study the effects of drugs in vitro. This should be extended to studies of solid tumors in vivo (it is already used for leukemia and in preliminary studies of some solid tumors), and to studies of drug effects in patients. Flow cytometry of semen and bronchial lavage specimens appears very promising for monitoring environmental pollutants. There was considerable speculation and much enthusiasm for the potential value of monoclonal antibodies as markers, but more caution about complementary nucleic acid sequences as markers. There also was much discussion about measurements of cell functions, (i.e., with Rhodamine-123 which is taken up by mitochondria) with dyes to measure cell membrane potential, and ' with fluorescence-tagged steroids to measure steroid receptors. It was concluded that the next several years will see very rapid advances in clinical and biological applications of both flow cytometry and high-resolution scanning cytometry.


SEMINAR AGENDA AND PARTICIPANTS

(1) SEMINAR ON IMMUNOGENETIC ANALYSES OF THE EXPRESSION OF TUMOR ANTIGEN
AND RESPONSES TO TUMORS
Osaka, Japan, November 29, 1982-December 1, 1982

AGENDA

PARTICIPANTS

UNITED STATES
Dr. Steven Burakoff
Dana Farber Cancer Institute
Boston, Massachusetts

Dr. Mark I. Greene
Harvard Medical School
Boston, Massachusetts

Dr. Richard J. Hodes
National Cancer Institute
Bethesda, Maryland

Dr. Stanley Korsmeyer
National Cancer Institute
Bethesda, Maryland

Dr. Susumu Tonegawa
Massachusetts Institute of Technology
Boston, Massachusetts

Dr. Ellen Vitetta
University of Texas Southwestern Medical School
Dallas, Texas

JAPAN
Dr. Shigeyoshi Fujimoto
Kochi Medical College
Kochi, Japan

Dr. Hiromi Fujiwara
Faculty of Medicine
Osaka University
Osaka, Japan

Dr. Yoshiyuki Hashimoto
Tohoku University
Sendai, Japan

Dr. Toshiyuki Homaoka
Faculty of Medicine
Osaka University
Osaka, Japan

Dr. Tadashi Kishimoto
Osaka University
Osaka, Japan

Dr. Noboru Kuzumaki
Hokkaido University
Sapporo, Japan

Dr. Masaji Okada
Osaka University
Osaka, Japan

Dr. Ko Okumura
Tokyo University
Tokyo, Japan

Dr. Kazuo Sugamura
Kyoto University
Kyoto, Japan

Dr. Masaru Taniguchi
Chiba University
Chiba, Japan

Dr. Tsuyoshi Uchida
Osaka University
Osaka, Japan

Dr. Takeshi Watanabe
Saga Medical College
Saga, Japan

Dr. Mitsuaki Yoshida
Cancer Institute
Tokyo, Japan



(2) SEMINAR ON GENE TRANSFER IN ANIMAL CELLS AND ACTIVATION OF CELLULAR ONCOGENES
East-West Center, University of Hawaii, Honolulu, Hawaii
January 17-19, 1983

AGENDA

PARTICIPANTS

UNITED STATES
Dr. Moses Chao
Columbia University
New York, New York

Dr. Geoffrey M. Cooper
Dana Farber Cancer Institute
Boston, Massachusetts

Dr. Barthol Sefton
Salk Institute
San Diego, California

Dr. George F. Vande Woude
National Cancer Institute
Bethesda, Maryland

Dr. Channing Der
Harvard University
Cambridge, Massachusetts

Dr. William S. Hayward
Rockefeller University
New York, New York

Dr. Michael Wigler
Cold Spring Harbor Laboratory
Cold Spring Harbor, New York

JAPAN
Dr. Hiroshi Amanuma
Cancer Institute
Tokyo, Japan

Dr. Yoji Ikawa
Cancer Institute
Tokyo, Japan

Dr. Masahiro Ishiura
National Institute for
Basic Biology
Okazuki, Aichi-ken, Japan

Dr. Yoshio Okada
Osaka University
Osaka, Japan

Dr. Takeshi Seno
Saitama Cancer Center
Research Institute
Ageo, Saitama-ken, Japan

Dr. Eiichi Soeda
National Institute of Genetics
Mishima, Shizuoka-ken, Japan

Dr. Tadatsugu Taniguchi
Cancer Institute
Tokyo, Japan


(3) SEMINAR ON "CYTOLOGY AUTOMATION"
Memorial Sloan-Kettering Cancer Center, New York, New York
February 20-22, 1983

AGENDA

PARTICIPANTS

UNITED STATES
Michael Andreeff, M.D.
Memorial Sloan-Kettering Cancer Center
New York, New York

Peter H. Bartels, Ph.D., PIAC (Hon.)
Dept. of Microbiology
University of Arizona
Tucson, Arizona

Kenneth Castleman, Ph.D.
Jet Propulsion Laboratory
Pasadena, California

Carlos Cordon-Cardo, M.D.
Memorial Sloan-Kettering Cancer Center
New York, New York

Don Evenson, M.D.
Memorial Sloan-Kettering Cancer Center
New York, New York

Paul Higgins, M.D.
Memorial Sloan-Kettering Cancer Center
New York, New York

Leopold G. Koss, M.D., FIAC
Department of Pathology
Montefiore Medical Center
Bronx, New York

Myron R. Melamed, M.D., FIAC
Department of Pathology
Memorial Sloan-Kettering Cancer Center
New York, New York

Lloyd John Old, M.D.
Sloan-Kettering Institute for
Cancer Research
New York, New York

Arline Redner, M.D.
Department of Pediatrics
Memorial Sloan-Kettering Cancer Center
New York, New York

Dorothy L. Rosenthal, M.D., FIAC
UCLA Center for Health Sciences
Los Angeles, California

Lisa Staiano-Coico, M.D.
Memorial Sloan-Kettering Cancer Center
New York, New York

John A. Steinkamp, Ph.D.
Los Alamos Scientific Laboratory
Los Alamos, New Mexico

George L. Weid, M.D., FIAC
University of Chicago
Chicago Illinois

JAPAN
Iwao Nishiya, M.D., FIAC
Department of Obstetrics & Gynecology
Iwate Medical University
Morioka, Japan

Kazuhiro Nomura, M.D.
Department of Neurosurgery
National Cancer Center Hospital
Tokyo, Japan

Manabu Takahashi, M.D.
Department of Pathology
Yamaguchi University
School of Medicine
Yamaguchi Pref., Japan

Masayoshi Takahashi, M.D., FIAC
Department of Pathology
Gifu University School of,Medicine
Gifu, Japan

Shigero Takamoto, M.D.
Laboratories of Chemotherapy and
Experimental Pathology
Aichi Cancer Research Institute
Nagoya, Japan

Noboru Tanaka, M.D., DDS, FIAC
Chiba Cancer Research Center
Chiba, Japan

Yoshio Tenjin, M.D., FIAC
Sasaki Institute
Tokyo, Japan

OTHER PARTICIPANT
Chester J. Herman, M.D., Ph.D., MIAC
Inst. Voor Pathologische Anatomie
Universiteit Nijmegen
Nijmegen, The Netherlands