TPC NEWS Winter 1990

TPC NEWS Winter 1990, Vol.9, No.1 (Whole Number 16)

(English Summary)

Page-3 Essay: A Visitor from the U.S.S.R.

In the last issue of TPC NEWS, I expressed my opinion on international exchange at our center, stating that we had not yet had guests from the U.S.S.R.. But just a little before the release of the NEWS I received a letter from Dr. Lapin, the Director of the Institute of Experimental Pathology and Therapy, Academy of Medical Sciences in the Soviet Union. The letter told of his plans to visit our center, which materialized on July 18th. Of course, Dr. Lapin was the first Russian scientist who has come to our center. And his visit was such a valuable event for us that we held a meeting to hear his lecture, under the auspices of the Researchers Association of National Institute of Health (NIH) of Japan.

I once had a Russian friend, Dr. S. G. Dzagurov, who was the director of the Tarasevic State Institute for the Control of Medical Biological Preparations, in the USSR's Ministry of Health. I first met him at a meeting of the scientific group on "Health Aspects of the Supply and Use of Nonhuman Primates for Biomedical Purposes", which was held by the World Health

Organization in Geneva in 1970. After that, we began to exchange letters. We frankly wrote opinions to each other on various matters, involving not only scientific problems, but also social and political affairs. He told me in his letters sometimes that he had an intention of visiting Japan. But, several years ago, before his visit could be realized, he passed away. 　I mourned his death deeply.

On the other hand, I have known of Dr. Lapin's high reputation since the 1960s. I once planned to translate "Nonhuman Primates for Medical Science" edited by him into Japanese, although the plan did not materialize. He was the researcher I always wanted to meet. Moreover, Dr. Lapin's Institute, built in Sukhumi, Georgia, in 1927, is the oldest research facility for laboratory primates in the world. In the 1960s, when the National Institute of Health (NIH) of America established seven regional primate research centers, it sent an investigative committee to the Institute in Sukhumi. The information given by the Institute was a very good guide for the NIH.

On the train from Tokyo to TPC, Dr. Lapin and I enjoyed our conversation, which was in English but also sometimes involving Russian words. I was surprised to hear that Dr. Dzagurov was his relative. And, just around that time, the ethnic struggle in the Republic of Georgia was reported to be escalating, so Dr. Lapin seemed to be worried about the safety of his family left in Sukhumi.

The aim of his visit was to learn of TPC's present activities and to promote exchanging information and studies. As well, he wanted to see TPC's facilities, which he had often heard about, since a plan to improve and enlarge his institute is now under way. It pleased me to hear Dr. Lapin express his admiration for the precision and effectiveness of our management of monkeys and laboratories.

He began his lecture, which was presented in beautiful Russian, with the following humorous introduction: "My institute has 8,000 primates, consisting of 7,000 nonhumans and 1,000 humans......, and is aiming at having 10,000 or more animals in the future.......". One thousand newborns have been born there every year, and studies of experimental oncology, infectious diseases, higher nervous activities, endocrinology, aging, and space biology have been actively carried out with the monkeys. The most interesting information for me was that 400 leukemia/lymphoma cases had already been found in the monkeys kept in breeding colonies. His two-and-a-half-hour lecture was full of interest for all of us.

Dr. Lapin born in 1921, is 68 years old. He is a sturdy, active person as well as a person of gentleness and sincerity. As he and I said good-bye to each other, we both wished a steady, continuing exchange between our two institutions.　Fortunately for us, we will be able to meet again in the 13th International Primatological Society Congress which will be held next July here in Japan.

Page-4 Breeding Topics: Utilization of wild　breeders ---On parity, abortion, stillbirth and nursing---

In 1977, the year before the establishment of TPC, we began selecting breeders from the wild cynomolgus monkeys imported, continuing this work until February, 1982. We selected almost the same number of breeders from three different monkey groups according to the country of origin, Malaysia, Indonesia and The Philippines. The numbers of the breeders at the end of the year are shown in the Table 1.

From 1980 to 1984, we were holding more than 500 wild breeders at the end of each year. 　After that period, the number has gradually decreased as the alternation of generation to F1 breeders has progressed. The female breeders who have been used since 1985 are the F1 generation produced through the Rotation Line Breeding System of TPC.

　　Pregnancies for the wild breeders totaled up to 2292 in number(See the Table 2). One of the breeders had ten parities for ten years. This is a rare case at TPC. The mean rate of fetal (RFL) at TPC was 13.2%. The rate can be said to be fairly low. The RFL depends on the species of monkeys and breeding conditions, being generally around 15% in indoor breeding colonies and occasionally more than 30% in outdoor and group feeding colonies. This comparatively low RFL at TPC is a result of our persistent and strict care and management of the monkeys. Although the number of the breeders having nine and ten parities is too small to lead a definite conclusion, the rate of fetal loss with nine and ten parities seems to be higher than those of the seven and eight parities. This is a noteworthy tendency.

　　　Successful nursing rate for the wild female breeders was more than 95%. This figure was natural, because we selected wild female breeders from animals showing clear development of nipples which can be regarded as an evidence of having nursed babies. The causes f the 5% which defaulted nursing were (1)immediately after Caesarean section, (2) difficult deliveries such as cephalic delivery and (3)functional troubles and anomalies such as abnormal weight loss and rectal prolapse. No cases were due to a defect in nursing ability. At present, the wild breeders seem to be more fertile than the colony-bred breeders at TPC.

Page-5 Serodiagnosis of Herpesvirus B by the Dot Immunobinding Assay(DIA)

　　　Since the opening of TPC, we have periodically carried out a serological survey on several antibodies to monitor viral infection at our monkey colony. Herpes B virus (HBV), in particular, is indigenous to Macaca sp., and known to be very hazardous to humans, ranking as Class IV of the criteria for biohazardous microorganisms. Recently, in the U.S.A., some fatal cases of HBV infection have been reported (two cases in 1987 and one case in 1989). The virus has a common antigenicity with HSV-1. Therefore, we had detected HBV antibody by the complement fixation test (CF), in which inactivated HSV-1 was used as an antigen. Recently, Dr. Kalter and Dr. R. L. Heberling of the NIH and WHO Collaborating Center for Reference and Research in Simian Virus, San Antonio, Texas, U.S.A. developed the Dot-Immunobinding Assay (DIA), which can detect HBV antibody more rapidly and sensitively than the CF test. Now, we are using this DIA to monitor our monkey colony.

　　The Dot Immunobinding Assay (DIA) can be said to be a kind of Enzyme-Linked Assay (EIA) using Western blot(WB). In the standard WB method diluted antibody solution is reacted directly to nitrocellulose sheets, whereas in the DIA method filter papers submerged in diluted antibody solution are applied to nitrocellulose gridded sheets(88 x 88 mm) pre-dotted with antigens. DIA allows a simultaneous testing for various antibodies with many serum samples.

　Cultural fluid of HBV inactivated with psoralen and ultraviolet irradiation is used as antigen. Following three cycles of freezing and thawing, the fluid is centrifuged at 2000 rpm for 30 minutes. One of the supernatant is dotted on a nitrocellulose gridded sheet. After the antigen-antibody reaction, the sheet is allowed to react with the second antibody labeled with alkaline phosphotase. As the result, a red spot appears at dotted sites if the sera contain HBV antibody. HSV-1 and uninfected cell cultures are used as control antigens. If a specimen is HBV-positive, the dots of HBV and HSV-1 change red. For identification of the virus in such a case, the tested serum is absorbed with cultural fluid of HBV or HSV-1 until either one of the colored dots disappears. The figure shown below is an example.

　Thirty-two sera from 16 wild-originated monkeys of our center, which had been positive for HSV-1 by the neutralization and CF tests, but were judged to be positive with 4 to 10 times intensity for HBV by this DIA. We would like to efficiently use DIA so that we establish HBV-free monkey colony.

Page-6 Introduction of Facilities: Serum Bank

　　TPC's serum bank is said to be a system of storing sera rather than a facility. The sera obtained from monkeys are aliquoted, labelled with stock number of eight figures, addressed and stored in a deepfreezer at -80 C. Until 1982, we managed the serum bank by a handwriting system. But, it has been included in a computer system since 1983.

　　The system of this bank consists of registering the data of the stocked sera, checking out the data and searching for the address of the sera required to be used. Now, about 40,000 specimens, including the sera obtained from imported wild monkeys at quarantine, those from monkeys at the time of periodical checkups done every two years and those from monkeys suffering from diseases, are stocked in this bank.

　　　The sera obtained from the people working with the monkeys when they were employed by TPC, have been stored, too. 　These sera are helpful to check the health condition of the people. This bank with four deepfreezers has a capacity to store about 70,000 specimens.

　　We use these sera for epidemiological, genetical and biochemical surveys, as well as, as criteria for some experimental data. For example, when ATL-like leukemia by STLV-1 infection was found in an African green monkey, we could trace the monkey back to six years before to find whether the monkey had been infected with STLV-1 before its arrival at TPC. In addition, we could offer the sera of wild monkeys that were not yet sensitized to cedar pollen, because before they were imported into TPC they had inhabited the area where no cedar trees were indigenous.

　　Not a few sera have not yet been used. We hope that this bank's sera will be utilized effectively in various fields.

Page-7 A Special Article: The major histocompatibility complex of laboratory primates

The cynomolgus monkey is used as a laboratory animal much more frequently than other nonhuman primate species, except for the rhesus monkey. In Europe and the U.S.A., difficulties in importing rhesus monkeys have increased the use of cynomolgus monkeys. In Japan, this species is used by far the most, in comparison to other nonhuman primates. Coincidentally, since it has become clear that MHC(major histocompatibility complex) plays an important role in self nonself recognition system with T cell receptors, elucidation of the MHC system of cynomolgus monkeys is needed in primate immunology. Further, through the development of molecular immunology of HLA and H-2, attention has been paid to primate MHC from the view point of evolution and disease.

In November 19-22, 1989, I attended the symposium "The primate MHC: Implication for evolution and disease" in Oegstgeest near the University of Leiden, the Netherlands. Here, I would like to relate recent information from primate MHC studies, along with a report of the symposium.

[1] Nonhuman primate MHC

The history of MHC was started as a result of tumor grafting in mice in the early 1900's. In this regard Little first described the basic genetic principles governing histocompatibility. With humans, Medawar(1946) first showed that graft rejection between unrelated individuals is implicated with the immunological system.

The MHC region is important because of its biological and immunological function and its relation to a system of self - nonself recognition. In addition, this region maintains extreme genetic polymorphism. Therefore, we can use this region as a good indicator of genetic control and monitoring of experimental animals and of genetic evaluation of population diversity of wild animals. In the mid 1960's, the need for laboratory primate models of human diseases increased with the development of the studies of renal transplantation. At that time, the studies on MHC of the rhesus monkeys were started in the Netherlands and the U.S.A.. In 1971, the first primate MHC workshop was held at Rijswijk, where TNO Primate Center is located. Twenty researchers from 9 laboratories met there for the purposes of identifying reagents, defining antigens and getting international agreement on nomenclature of primate MHC. n this conference it was decided that confirmed alleles were designated as Rhw.

However, the report using the designation has not been found. I heard that the second workshop was held in the U.S.A. but did not know the details because of the absence of report. MHC of Japanese monkeys was reported in the Congress of Japan Immunology Society (1982) by A. Noguchi. However, genetic evidence was not sufficient because there were very few well documented pedigrees for family analysis. For this study, he could use only colonies having undecided fathers and wild populations which are not suitable for a family analysis.

[2] The symposium of primate MHC in the Netherlands

Few distinguished reports of primate MHC has appeared because of delayed establishment of an artificial breeding system and the difficulty of importing after the Washington Convention on International Trade in Endangered Species of Wild Flora and Fauna in 1973. Only a few primate centers having large or many breeding colonies could survive with regard to primate MHC research because MHC analysis requires many monkeys and their families.

At the symposium "The primate MHC: Implications for evolution and disease", 50 researchers met at a corner of the Castle Oud Poelgeest in Leiden. (Number of participants; the Netherlands 22 persons, F.R.G. 8, U.S.A. 7, U.K. 3, Spain 2, Australia 1, France 1, Sweden 1 and Japan 1.)

First, Dr. Balner, who was retired from the director of TNO Primate Center talked about the history of primate MHC. Briefly, the studies of MHC of rhesus monkeys, chimpanzees and baboons were conducted at first in 7 laboratories. At present, however, only TNO Primate Center can continue to study this project. Thus a comparison of evaluation of antisera was not necessary. Other primate centers could not continue MHC studies because of the difficulty to maintain and breed monkeys for this project.

Why is it possible for TNO Primate Center to type RhLA antigens of all rhesus monkeys of the Center and the monkeys from other centers on request? The answer is the presence of two key men -- Dr. Balner of TNO Primate Center and Dr. van Rood of the University of Leiden, who embarked on a long-standing and productive collaboration in transplantation immunogenetics. Besides, Rijswijk and Leiden are very close to each other. And the Netherlands is a so-called transport center for animals in Europe. Van Rood is one of the pioneers of the studies of the HLA system.

Since nonhuman primates are outbred and phylogenetically close to man, RhLa typing was demanded for the development of transplantation immunogenetics in the rhesus monkeys as a model of HLA. And these studies paved the way for an organization like Eurotransplant ( Organ Transplantation ). The other important pre-clinical studies in nonhuman primates involve testing of new immunosuppressive modalities, such as pre-transplant bloodtransfusions.

The program of this symposium was as follows:

[ From Apes to Human...the way of MHC polymorphism ]

by J. Klein.

[ Typing for ChLa class I polymorphism using human alloantisera ]

by A. van Leeuwen

[ Comparison of class I MHC genes in human and apes ]

by P. Parham

[ Evolutionary analysis of allelicdiversity and haplotype combinations at the HLA class II DQ, DR, and DP loci in primates using DNA amplification ]

by U. Gyllensten and H. Erlich

[ Polymorphisms and evolution of the MHC loci]

by M. Nei

[ Matching for the MHC ]

by J.J. van Rood

[ MHC-class II polymorphisms in primates: from gene to protein to function ]

by R.E. Bontrop

[ Ancestral haplotype in nonhuman primates with particular reference to central MHC genes ]

by R. Dawkins

[ PCR typing ]

by H. Erlich

[ MHC immunity and immunopathology ]

by R.R.P. de Vries

[ MHC class II and cell activation ]

by D.J. Charron

[ Successful treatment of EAE in rhesus monkeys with MHC class II specific monoclonal antibodies ]

by M. Jonker.

We had more 9 presentation from the floor, including my own report [Leukocyte antigens of cynomolgus monkeys ]. I think the finest topic of this symposium was the report by Dr. Nei. Listening to his lecture, I felt that MHC studies should be reconsidered from the view point of population genetics dealing with data obtained by results of the recent molecular immunological analysis of homology between men and mice. On the last night of this symposium, nomenclature of markers of primate MHC were discussed, It was agreed that, for example, in the case of chimpanzees, a marker is named [ MHC Patr-Dr8 ].

From this conference, I keenly think that there is a necessity to establish a genetically-controlled breeding colony of laboratory primates which have many unrelated monkeys as well as many pedigreed families, for studies on primate MHC and primate immunology. A deadlock of primate MHC studies in Europe and U.S.A. resulted from a defect of biomedical research system that was not able to carry out breeding under genetic control.

I am sure that, since TPC has been breeding and maintaining monkeys under the policy of gene preservation, only such a center can develop further primate MHC study. My feeling is that this is a historical new breath in the air similar to the Breakdown of the Berlin Wall.

Page-11 A Comment on My Research: Recollections of my 37-year study life and hopes for the future activities of TPC

TPC Director, Dr. Shigeo Honjo will retire from his present position at the end of this coming March. The Editorial Board of TPC News asked him, therefore, to write a summary of his 37-year research history for publication in TPC News. Dr. Honjo kindly accepted the request, and in "A Comment of My Research", he has recollected memories of his research life, and has expressed his hopes for TPC's future activities.

Dr. Honjo has divided his past research life into three periods. The first was from April of 1953 through March of 1961, during which time he worked for Ibaraki University, after graduating from the University of Tokyo. The second period began in April of 1961, when he moved from Ibaraki University to the Department of Veterinary Science at the National Institute of Health, continuing there until March of 1978, just before the establishment of TPC. The third period was from April, 1978, when TPC was established, until today.

Ibaraki University, where Dr. Honjo worked during the first period of his research life, is a local university established along with the new higher education system of Japan after the end of the second World War. At that time there were many young and active scientists at Ibaraki University, They were all filled with enthusiasm for developing research and education on the basis of a new viewpoint, democracy. Although research facilities and equipment for research were quite poor at the newly-established local universities because of nation-wide poverty brought on by defeat in war, Dr. Honjo was able to develop his research activities freely, thanks to the democratic atmosphere of the university. One of his most impressive works during this period was a paper he published with Dr. Imamichi, regarding experimental results to contradict the endocrine theory of salivary glands which was popular in the field of medical endocrinology in Japan at that time. They proved that the established theory contained not only methodological faults caused by the restrictions of the times, but also cognitive faults, which viewed corelationships in physiological phenomena as causal relationships. Moreover, such an erroneous theory had been reproduced extensively under semi-feudalistic (anti-democratic) conditions in the Japanese academic world before the end of World War II. At any rate, American scientists also later denied the endocrine theory of salivary glands. Dr. Honjo learned from this research experience in his younger days, that developing scientific work requires a strictly critical attitude and resistance against incorrectly established authority.

Around the time when Dr. Honjo moved to the National Institute of Health in 1961, a widespread epidemic of acute poliomyelitis was hitting Japan. The public was agitated about the development of polio vaccine and the establishment of social preventive system. In order to protect children's health, many mothers joined a strong social movement demanding vaccinations and the promotion of polio-research. Most scientists responsible for people's health worked hard to honor their social responsibility. Dr. Honjo, for one, was working on the care and management of the monkeys indispensable for the production and testing of polio vaccine, In those days, his division at N.I.H. imported about 1500 monkeys per year. Quarantine and health care for those imported wild monkeys was carried out at a monkey facility, which was not so well constructed or equipped as today's TPC facility. He says that sometimes more than 50% of the new arrivals died during the first 2 to 3 weeks after importation. In those days, the term "biohazard" had not yet appeared, Dr. Honjo autopsied a lot of dead monkeys with care nonetheless, to guard against B virus infection. Accumulating and analyzing abundant data and experience, he made efforts to build up a concept of "monkeys as laboratory animals" in a paper which was published in 1965 in Seibutsu-kagaku(Biological Science), a Japanese language scientific journal. That concept is essentially the same as the one which still constitutes the principle of the TPC.

When Dr. Honjo began to work at N.I.H., he thought that it was necessary and indispensable to succeed in breeding under artificial feeding conditions, in order to establish monkeys as laboratory animals. In 1962 he and his colleague, the late Dr. Fujiwara, began basic research on breeding the cynomolgus monkey. Their studies on breeding developed at a steady pace, even though the scale of their breeding colony was small. Fortunately, the necessity of breeding nonhuman primates as laboratory animals which Dr. Honjo so strongly advocated, was recognized in 1967 by the Future Plan Committee of N.I.H. scientists. From that time until the establishment of TPC, Dr. Honjo and his research associate were able to obtain infant cynomolgus monkeys of the 3rd filial generation.

Dr. Honjo was also aware of the importance of research into diagnosis and control of naturally occurring diseases among nonhuman primates. When imported wild monkeys exclusively were employed as experimental animals, one of the most troublesome diseases encountered was bacillary dysentery. It occurred very frequently at almost all nonhuman primate facilities in the world. Therefore, Dr. Honjo chose "Shigellosis in the cynomolgus monkey" as one of the main research subjects in the second period of his research life, and worked most closely with Dr. Takasaka during the course of this investigation. The extent of their studies was wide, ranging from an epidemiological survey on natural Shigella infection among wild cynomolgus monkeys, to the establishment of an experimental model of shigellosis in the cynomolgus monkey. After success in some small scale experiments, Dr. Honjo organized a collaborative research group consisting of scientists from the Department of Veterinary Science, the Department of Pathology and the Department of Bacteriology at N.I.H.. They were all excellent colleagues dedicated to their collaborative studies. Their efforts resulted in the successful establishment of shigellosis models in the cynomolgus monkey models through such various experimental infection routes as oral, intra-cecal and anal routes. Obviously, those cynomolgus monkey models of bacillary dysentery are very useful, because of their close similarity to human bacillary dysentery. For example, killed Shigella oral vaccine that was developed under the auspices of the Ministry of Health and Welfare just before the 1964 Tokyo Olympics, was tested for its efficacy by the use of this cynomolgus monkey model of shigellosis. Results of the test demonstrated that the vaccine was not effective for the prevention of shigellosis. Dr. Honjo was very happy with his research activities in those days.

With the 1970's, Dr. Honjo had to spend fairly long hours making a basic plan for the present TPC. Dr. Cho had been working at Dr. Honjo's laboratory since 1968, cooperated closely with Dr. Honjo on the plan. It gradually became concrete with their efforts as they set out a unique Japanese idea, as well as absorbed and integrated the many experiences and results obtained by American Primate Research Centers. It was in this way that TPC was established and began to function in 1978.

On the basis of the above-mentioned experiences in the second period of his research life, Dr. Honjo became convinced that it was fundamentally important to know that there are many problems in the field of laboratory primates, and that they should be solved using pragmatic lines of thought. Furthermore, he believed that a basic and steady research approach was needed for the resolution of such problems. And it made him happy to know that his laboratory was always filled with an atmosphere of freedom, which is one of the most necessary conditions for the sound development of academic studies.

The third period of Dr. Honjo's 37-year research life began with the inauguration of TPC in April of 1978. When TPC's plan first proposed, Japan's economy was at the peak of its development. Economic conditions, however, had more or less darkened by the time TPC started functioning. And public opinion seemed to want small scale government, because public finances were tight. Therefore, TPC was faced with a national policy of a steadily-reducing annual budget.

When Dr. Honjo began to work at TPC, he felt that the realm of research there should include at least six fields: reproductive physiology (especially, endocrinology), genetics and breeding science, immunology and clinical pathology, pathogenic microbiology, nutrition science and behavior science. In addition to these research fields, he wanted three sections to be organized at TPC: a section for developing and improving primate clinic and feeding-related techniques for primates, a section for collecting and distributing organs and tissues obtained from monkeys autopsied at TPC. Dr. Honjo's original plan has not yet materialized completely, however, because of continuing financial restrictions. Nevertheless, Dr. Honjo has organized many research teams on different subjects, always encouraging TPC people to fully demonstrate their abilities.

According to Dr. Honjo's often-published opinion, the research activities at TPC extending over the six scientific fields and three work sections described above should stand on three basic standpoints. The first deals with the practical problems which occur constantly at the front of feeding more than 2,000 monkeys. Such problems include establishing and improving feeding equipment, clinical diagnostic methods, therapeutic and prophylactic measures, developing a breeding system that is useful for genetic control, and improving a rearing system for the induction of normal sexual behavior in colony-born monkeys, etc.. Studies dealing with these practical problems are undertaken mainly through the empirical or retrospective method. About 20% of the original articles published by TPC personnel have been categorized as studies carried out from this standpoint. The results obtained suggest numerous problems, which should be investigated from a second standpoint.

The second standpoint is to clarify the biological characteristics of nonhuman primates through a multifaceted approach. Studies classified as belonging to the second category mainly employ experimental methods, but also include survey and testing methods. This category of TPC's research activities includes, for example, analyses of genetic polymorphism of blood groups, MHC antigens, serum proteins and mitochondrial DNA, determination of hormones pertaining to reproductive mechanisms, analyses of relationships between aging or fetal development and immunological factors, biometrical analyses of normal body growth, and so on. In addition, microbiological and pathological studies on naturally occurring diseases are also conducted from the second standpoint. The results of these basic studies will directly or indirectly contribute to the resolution of many practical problems occurring at TPC's nonhuman primate colony. At the same time, the studies may result in the development of nonhuman primate models of human diseases. Dr. Honjo estimated that about 60% of the original articles published by TPC were based on the second standpoint.

He has stressed advantages of researching at TPC from the second viewpoint. First, TPC maintains many laboratory-bred monkeys whose ages are exactly known ranging from a conception date to a very old age. Secondly, TPC has many monkeys of successive generations whose consanguinity is well defined. These conditions can never be satisfied if scientists have no choice but to use wild-originating monkeys alone for their studies and if only a few animals are kept at their research facilities. Dr. Honjo strongly wants that TPC's people will make efficient use of these advantageous conditions for their further studies. The third standpoint of TPC's research activity advocated by Dr. Honjo is to make conscious efforts to demonstrate the usefulness of nonhuman primates as laboratory animals in various research fields, in particular, the development of useful nonhuman primate models of human diseases. The number of paper written from this standpoint constitutes about 20% of the entire body of TPC publications.

In the opinion of Dr. Honjo, the first step of research carried out from this third standpoint is to thoroughly perform clinico-pathological examinations of naturally occurring diseases. And then the character of a disease occurring among monkeys should strictly be compared with that of a corresponding diseases in man. Next, efforts are made to produce an experimental disease model, using some factors presumed to be etiological in the naturally occurring disease of the monkey concerned. In some cases, a causative factor that was determined to a certain extent in humans is experimentally applied to healthy monkeys for the production of an animal model of the human disease. Moreover, there is a way in which scientists employ some factors that are not directly implicated in naturally occurring diseases, but can induce symptoms and/or pathological lesions similar to those observed in naturally occurring diseases (for example, MPTP-induced Parkinson's disease model and Streptozotocin-induced type-I Diabetes mellitus in macaques). If a genetic background is associated with a naturally occurring disease, a patient breeding effort should be made to produce so-called honozygotes. Dr. Honjo also points out the necessity of developing refined techniques concerning the production of early embryos, fetuses and infants that have a transferred gene of a certain genetic disease.

Dr. Honjo has always sought the collaboration on this research activity, of scientists from various disciplines at other research institutions, because he would like to see more official governments-accredited scientists working at TPC. And he has stressed that further efforts should be made by TPC's researchers to develop nonhuman primate models of brain and nervous system disorders, geriatric diseases and genetic diseases.

During Dr. Honjo's 12-year career in Tsukuba, the most pleasant thing for him was that he heard reports every day of new developments and accomplishments in breeding and rearing many monkeys. He always encouraged the people who were working in the animal facilities as well as in the laboratories, to perform their work with a scientific attitude and to publish the results of their work. Drafts of every paper written by TPC's people were strictly reviewed and amended by Dr. Honjo.

Dr. Honjo has also published his opinions about the social movement against the use of animals for biomedical experiments and testings. He has stated unequivocally that animal experimentation is an important and indispensable method in research which is necessary for ensuring and promoting human health and welfare.

He added that the so-called alternative method to animal experiments is merely one of many research methods, which can never replace animal experiments in the true sense of the words. These days, some believe alternative methods are unquestionable. But Dr. Honjo believes that an independent course of alternative methods alone is more dangerous even than that of animal experiments which are carried out without regard to the relationship between the human side and that of the animal.

Dr. Honjo finished writing his recollections on October 31, 1989, expressing his hearty thanks to all the people who worked with him for their invaluable collaboration and advice during the past 37-years.

Page-17 Case Report: Ovarian granulosa cell tumor in a cynomolgus monkey

Statistics by the Japan Society of Obstetrics and Gynecology show that 4.6% of solid ovarian tumor cases were ovarian granulosa cell tumor cases. It has been reported that, in human beings, this disease occurs in both young and middle aged individuals with two peaks of incidence, below 10 and at 40 to 50 years of age. This cynomolgus monkey case is the first ovarian tumor case we experienced at TPC.

Case : A female cynomolgus monkey of Philippine origin. Her estimated age was 15 or more. She had five parities at TPC, but had been amenorrheic for six months before death.

Autopsy findings: The right ovary was remarkably enlarged (12x17x7 cm) and gray in color. Its surface was smooth, upheaving partly (Photo. 1). Torsion of pedicle was observed. The cysts of various sizes were observed in the tissue-sections routinely prepared. They were filled with a gelatinous, faintly yellow fluid. The left ovary had no significantly abnormal findings.

Histologically, many Call-Exner bodies were recognized: Amorphous eosinophilic substances surrounded with tumor cells were observed in small round cavities. The shape of the nuclei of the tumor cells were oval, and their cytoplasm was faintly dyed.

Page-18 Report on the Symposium "The Role of Laboratory Primates in Aging Research"

The 36th General Assembly of The Japanese Association of Laboratory Animal Science(JALAS) was held in Tokyo from May 29 to 31, 1989, having a symposium titled "The Role of Laboratory Primates in Aging Research". The research on aging and brain structure and function will be the most important issues hereafter in the field of biomedical science. Six lectures, including that by Dr. Bowden of Regional Primate Research Center at the University of Washington, were presented in the symposium. They all demonstrated that the usefulness of nonhuman primates as an animal model in aging research.

The proceeding of this symposium will be issued in Experimental Animals (Jikken Doubutsu), the official organs JALAS, Vol. 39, No. 2.

Page-19 The Statement of the Vth Congress of Primate Society of Japan

For the establishment of breeding and supply system of laboratory primates in Japan---

Primate Society of Japan adopted unanimously a statement asking for the establishment of breeding and supply system of laboratory primates in Japan, on July 25th at the general meeting of its Vth Congress held in Tokyo from July 24th to 26th.

The statement was completed following the discussion and suggestion, which were made in the symposium on "Conservation and useful use of nonhuman primates in biomedical research" held on the day before the general meeting. This statement was sent or handed to governmental authorities concerned under the name of Dr. Masao Kawai, President of Primate Society of Japan.

Primate Society of Japan has been strongly concerned about the conservation of wild monkeys since its establishment, discussing the use of laboratory primates.

In the symposium on " The Present State and Problem of Laboratory Primates in Japan" held during the Vth Congress of PSJ, we affirmed the following recognition:

Laboratory primates play an important role for developing and maintaining human health and welfare.

When we use laboratory primates, we should give serious consideration on wild primate conservation.

The establishment of breeding and supply system is essential for both conservation of wild primates and the improvement of quality of laboratory primates.

The PSJ appeals the following four points to the public and organizations concerned.

(1) The use of wild primates in biomedical research and testing should be done in careful consideration for their wild conservation.

(2) Artificially bred and reared animals should be used in biomedical laboratories. Therefore, large scale breeding facilities be established.

(3) A large scale breeding and rearing of primates for laboratory use requires a great amount of money, and inevitably needs gene preservation and various basic studies. For establishing and improving such projects, political and financial support by nation is indispensable.

(4) The existing institutions and organizations related to laboratory primates should endeavor to make a flexible plan opened to the outside, and if necessary, to reorganize themselves to actualize the three points mentioned above.

Page-20 Overseas Topics: Financial aspect of the German Primate Center(DPZ)-II-

According to the Primate Report 24, March 1989, the financial state of the German Primate Center in 1988 was so good that the administrative director of DPZ told that 1988 had been an exiting year. Four figures show the DPZ's Total Income in 1988, Total Expenditures in 1988, Income Development 1977-1988 and Employees in 1988.

Page-21 A Memory of a Trainee: TPC and I ---TPC's door is always open to those students who want to study with laboratory primates---

At TPC, Miss F. who is a student of Tsukuba University's Master Course has been studying on obesity, metabolic diseases and immune function in cynomolgus monkeys. She describes in this memory why she decided to become a trainee at TPC and what she felt as a trainee.

First, she was deeply impressed with TPC's openness and kindness to those who want to study with laboratory primates, as well as, its excellent system for animal care and management. She will never forget what she learned at TPC, thanking all TPC people very much.

Page-21 Sketches from Animal Rooms: Thinking in a monkey colony

An animal technician Mr. N. tells that he works with monkeys at TPC, sometimes reminding of monkeys which appeared in old Japanese stories and ancient Indian stories such as Rahnayana. He must have much literary culture.

Page-22 Sketches from Animal Rooms: Cage-repairing

Mr. H., an animal technician reports on cage-repairing work at TPC.

Now, TPC has about 2,000 cages which are classified into several types. All of the cages are made of stainless steel. Therefore, they are rustless and clean, although they are considerably heavy. But, today, twelve years after TPC's opening, many damaged parts are seen in those cages. When the damage is not serious, we repair them by ourselves, 20 to 40 cages at a time, twice a month. Repairing the cages of TPC is fairly hard work, because the number of the cages too great.