TPC NEWS Vol.16, No.1 Summer, 1997
(Whole Number 27)
( English Summary)
Page-
3 Message from the chief director
of the Corporation for Production and Research of Laboratory Primates (CPRLP):
Current topics on infectious diseases
Dr. Toshiro Matsuura, the chief director of the
Corporation for Production and Research of Laboratory Primates (CPRLP),
describes in this note an aspiration
of strengthening CPRLPfs
commitment to the social changes and needs, reviewing several current topics
including O-157 and legionella outbreaks in Japan.
Page -4
Message from the new director of the TPC: Greetings
Dr. Akio Yamada was inaugurated as Director of the TPC, succeeding Dr.
Yoshikawa. Until the end of this
March, he had been the chief of the Laboratory of Mumps Virus and Vaccines,
the Department of Viral Disease and Vaccine Control, the National
Institute of Infectious Diseases (former NIH, Japan).
He says that he have committed with monkeys as an user of vaccine safety
tests so far, and this new position made him face with a new field for him.
Now he is urgently studying the TPCfs systems in both sides of hard and
soft. He wishes he will be able to contribute to further development and leap of
the TPC.
Page-5
Message from the former director of the TPC: Looking back over the events
at the TPC
Dr. Yoshikawa, former director, contributed this message to the editors
as his farewell greetings. He is
looking back over the 6 years he spent at the TPC, the second stage for the TPC,
picking up several major organizational events.
For
instance, in 1992, the tasks of the TPC was expanded. In 1994, the facility of Biosafety Level 3 (P3-Laboratory)
was built, and also the project of a research resource bank for the primates was
started, which implied that the TPC should have a function as a joint use
facility for research with monkeys.
The
year 1995 was a big year for the TPC. The
budget for the construction of the joint use facility for research by the
Ministry of Health and Welfare has been passed.
The working group to make the original plan for hard and soft wares of
the facility was started, and in 1997, supplementary budget for the construction
of the information center with accommodations was passed.
In these facilities, research by new technologies such as gene therapy,
and the studies of brain science and aging shall be carried out along with the
research for infectious disease models.
In addition to these events related to the TPCfs management, Dr.
Yoshikawa recollects the events in which he had good time with the TPCfs
people, such as a cherry blossom party in spring, a beer party in summer, and a
year-end party at the Mt. Tsukuba, as well as soft ball games held every year.
He says that it has been his great pleasure having a lot of time to
discuss with the people and having been in beautiful nature in Tsukuba.
Page-7 Research Project at the TPC:
Characterization of
peripheral blood CD4+ CD8+ double-positive lymphocytes in
cynomolgus monkeys
Hirofumi
AKARI
In this study, we analyzed peripheral blood CD4+ CD8+
double-positive (pDP) lymphocytes in adult cynomolgus monkeys (Macaca
fascicularis). Forty of 55
monkeys had more than 5% of pDP subpopulation (9.3 }
5.9%; mean }
SD) in peripheral blood lymphocytes (PBL) in contrast to low percentage of@pDP
cells in humans and mice. In cross-sectional study, the pDP cells were found to
increase in proportion with age.
To clarify whether pDP lymphocytes were immature precursors released from
thymus without prior differentiation, the expressions of CD8 chains and CD1b on
pDP lymphocytes were compared with those on thymocytes.
The pDP lymphocytes were CD8ż+Ŕ-
and CD1b-, while thymic DP lymphocytes were CD8ż+Ŕ+
and CD1b+, suggesting that the pDP cells are extrathymic T lymphocytes.
Furthermore, the pDP@lymphocytes
exhibited a resting memory T cell phenotype with CD2hi CD3+CD28- CD29hi CD49dhi
CD69- CD80lo. Taken together, adult
cynomolgus monkeys possess a unique pDP T cell subpopulation which express a
resting memory T cell phenotype. In
addition, similar phenotypic properties of pDP lymphocytes were distributed in
the spleen and lymph node, although the proportion was less in the spleen and
much less in lymph nodes than in PBLs.
Page
- 12 Research Project at the TPC:
Evaluation of anti-AIDS drugs in monkeys
Ryozaburo
MUKAI
Simian immunodeficiency virus (SIVmac) causes immuno-suppression and
diseases similar to those in AIDS patients, therefore, it provides potential
models for HIV infection and AIDS. Its
clinical and immunological features are as follows : 1) reduction of CD4 cells
and decrease in blastogenesis of peripheral blood mono-nuclear cells (PBMC),
2) opportunistic infection such as abscess formation (Streptococcus aureus) and
reactivation of Cytomegalovirus (CMV) and pneumonia (Puemocystis. carinii), 3)
anorexia, diarrhea and emaciation, 4)occurrence of AIDS encephalitis (fig. 1),
and 5) presence of long-term non-progressor.
In order to establish the methodology to evaluate anti-HIV drugs in
animal models, we examined
anti-viral activities in rhesus monkeys, by using 6-Chloro-2',3'
dideoxyguanosine (6-Cl-ddG). The
monkeys tested were consisted of 3 acute phase, 3 asymptomatic carrier (AC)
stage and one ARC/AIDS monkeys.
Administration of 6-Cl-ddG to the ARC/AIDS monkey resulted in remarkable
recovery in clinical signs and symptoms including weight gain, stop of
diarrhea and disappearance of neuropathy. The
increase in CD4 and CD8 cells and the decrease in plasma viral load were also
observed during and after the treatment in this case (fig. 2).
In
the acute phase monkeys, the appearance of antibodies to SIV, plasma viremia and
cellular viremia significantly delayed during the treatment (fig. 3).
In
the AC stage monkeys, the decrease in cellular and plasma viral load was
observed during administration of 6-Cl-ddG (fig. 4).
Therefore, the evaluation of anti- HIV drugs would be possible in rhesus
monkeys by the quantitation of cellular viral load using cocultivation and
plasma viral load using Reverse transcriptase polymerase chain reaction (RT-PCR)
in all stages of infection with SIV.
In the near future, the construction of chimeric virus (SHIV) of which
env and pol regions are derived from SIV is necessary to develop new anti-AIDS
drugs more efficiently.
Page16-Breeding
Topics: Nursing ability of laboratory-bred breeders
Takahiro ONO and Fumiaki CHO
The number of deliveries in laboratory-bred breeders has just exceeded
2,000 in the TPC. A total of
2,039 normal deliveries ( No. of multiparous breeders: 590 ) covers the
1st parity to the 11th parity at maximum (the 7th, 8th, 9th and 11th parities
were collectively dealt with as "the 7th parity and thereafter" in
this report owing to scarcity of the number of deliveries at each of these
parities ).
Fig. 2. The nursing ratios
by parity. The ratio at the 1st
parity, 52.2%, was significantly lower than that at the 2nd parity or
thereafter(p<0.05);the nursing ratio rose as parity increased .
Fig. 3. The nursing ratios
at the 1st parity by generation. No
significant differences were seen among the generations of
F1 to F3.
Fig. 4. The nursing ratios
at the 1st parity by breeding system, by which the deliveries were brought
about. No significant differences
were seen among the breeding systems.
Fig. 5. The nursing ratios
at the 1st parity by age . No significant differences were seen among the ages of 4 to 9
years (p<0.05).
Fig.6. The nursing ratios at the 1st parity by years. Technical advancement of animal technicians may have reflected on the nursing
ratio, which was improving up to 1989. Simian
varicella-like herpesvirus infection, which occurred in the TPC at the end of
1989 and resulted in the six month-suspension of reproduction and elimination of
a large number of infected animals, determined the breeding efficiency (lowered
the nursing ratios) thereafter.
The present report, the third report on the nursing ability of the
laboratory-bred female breeders in the TPC, validates the following results: (1)
the nursing ability of laboratory-bred breeders at the 1st parity is inferior to
that of wild-born breeders; (2) the nursing ability of the laboratory-bred
breeders is improved at the 2nd parity or thereafter; the nursing ratio rose to
90.9% (169/186)at the 5th parity, though it is still lower than 97.0%
(1973/2033) obtained in the wild-born breeders; (3) the improvement in the
nursing ability at the 2nd parity or thereafter implies that the first
parturition at an earlier age leads to the improvement in breeding efficiency at
the 2nd parity or thereafter; and (4) in order to breed monkeys with higher
nursing ability, it is needed to establish a new breeding/rearing system, which
will be actualized by the technical advancement of animal technicians.
Page-18
Laboratory Tests: Use of polymerase chain reaction (PCR) for diagnosis of
simian retrovirus / type D (SRV/D) infection
Toyoko
NARITA
Simian retrovirus/type D (SRV/D) is a group of viruses
having relatively close antigenecity, and macaque monkeys are infected
spontaneously. In 1980s, it was
revealed that this virus caused serious immunodeficiency in macaque monkeys, and
so far, immunodeficiency cases in eight different species of macaques have been
reported. Infection of this virus
in monkeys varies in its clinical signs. Its
inapparent infection has possibility to result in virus carriers having no
evident antibodies, and the carrier may become a cause of prevalence in
colonies.
In
order to produce the free colony from SRV/D infection, we have established a
method to detect the proviral DNA of SRV/D by using polymerase chain reaction (PCR).
The
primer was designed according to the conserved sequence of SRV/D-1 to 3.
Fig. 1 shows the results of nested PCR using standard SRV/D-1 and SRV/-tsukuba.
@
SRV/D
DNA were detected directly by PCR, using the peripheral blood mono-nuclear cells
(PMBC) co-cultivated with Raji cells for two weeks and for three weeks.
The results were shown in Fig. 2.
Fig.3
summarized the detection of SRV/D antibody by Western blot analysis (WS) and
proviral SRV/D by the PCR.
Page- 20 Case Report:
Hamartoma of the lung in a cynomolgus fetus
Ippei
SKAKIBARA
Hamartoma is a focal malformation that looks like tumor.
I have never seen any case report on hamartoma in nonhuman primates.
This is the first case in our Center.
Case: A cynomolgus
monkey fetus of 171 gestation days. The
fetus, whose mother had been diagnosed as placenta previa, died soon after
cesarean section.
Histological findings: Marked
edema were seen in the subcutaneous tissue of whole body. A huge tumor (28.5 g)
originated from the left upper lobe occupied the thoracic cavity (Fig. 1).
Tumor surface covered with membrane was smooth and milk-white in color.
Cross-section was evenly spongy.
No adhesion to adjacent organs such as the pericardinal cavity and chest
wall were observed. Histological
structures of the tumor were similar to those of the lung of normally developed
fetuses: glandular carcinoma-like lumens, epithelial cells of respiratory
apparatus ( alveolar, bronchial, and columnar ciliated epithelium), bronchial
cartilage, connective tissues and blood vessels were seen.
The luminal connective tissues were edematous (Fig. 2).
Aplasia of thymus was suspected since no thymus was seen by naked eyes
nor histological examination. The
liver was hyperplastic a little, of which surface looked uneven.
Atrophy and degeneration of liver cells are observed under the hepatic
theca. Pale yellow pigmentation was
seen in cytoplasma of the liver and Kupffer cells. Proliferation of collagen
fibers was seen.
Page-21 Clinography: gThis is the
TPC Clinich
Fumiko
ONO
Now we are introducing a computerized administration system of clinical
records. The data of our monkeys, such as the body weight, observation records,
family line, and mating records have already been computerized, but the clinical
records were hand- written on paper sheets.
Then, we began computerizing the clinical data three years ago.
Here I report several topics picked up from the clinical records.
The total number of treatment to the monkeys from April f96 to March
f97 was about 10.000. It means that 35% of the cynomolgus, 8% of the African green
and 11% of the squirrel monkeys of the TPC underwent to medical treatments.
As for the cynomolgus monkeys, most of the cases were diarrhea : about
40% of the treatments to them. The
second major case was a wound in a variety of degree from a serious one by
fighting to a slight one made by a cage. The
number of the monkeys underwent to the wound treatment was 161 (21%).
During mating period, in particular, it is not rare that female breeders
seriously suffer injuries from bite by males (Fig. 1, 2).
In that case, we stop mating immediately, and clean and sterilize the
wound with hydrogen peroxide, saline or iodine, and apply an ointment of
antibiotics as well as the one to promote tissue recovery.
Blood examination and transfusion are sometimes done to the animals according to their symptoms.
Fig. 1 shows a case that recovered from serious bite injuries in about 2
weeks. However, there were five
death cases by septicemia with pneumonia or myocarditis resulted supposedly from
bites.
The following disorders were also seen in the mating period: diabetes
resulted from lack of exercise and overeating, malnutrition and hypoglycemia
resulted from persecution by mating males, and anemia that seems to be related
to the viral diseases induced by the stress in the mating period.
Recently, the following six tumor cases were found out.
Case1:
A 3-year-old female cynomolgus monkey, suspected of chondroma at the left caput
femoris.
Case 2: A 3-year-female cynomolgus monkey, suspected of the lymphoma at
the right cheek (Fig. 3, 4).
Case 3: A-wild- male green monkey, suspected of the basal cell carcinoma
at the chest.
Case 4: A 16-year-old male green monkey, suspected of the hepatocellular
carcinoma.
Case 5: A 1-year-old male cynomolgus monkey with encephaloma.
He had a squint about one month before his death.
Case
6: A 13-year-old male squirrel monkey with lymphocyte leukemia.
The
occurrence of the disease are induced by many factors such as the environment,
pathogens or genetic factors. We must investigate the cause of diseases by not
only the investigation on each
animal but also by epidemiological and comprehensive survey using the database
as a colony record. Then, we can
prevent the spread of pathogens as soon as possible, and select valuable disease
models.
It
is needed to make a network to use the information effectively.
Any information and advice on the clinical medicine of the primate are
welcomed.
Page-23
Topics: Importation and quarantine of primates
Dr. Yoshikawa had been the chief of a study group of prevention of
zoonoses through primates, one of the Health Science Research Programs by the
Ministry of Health and Welfare in Japan (MHW).
This yearfs report of the study already have been submitted to the MHW.
The following is a brief summary of the report.
This yearfs object of the study group was to discuss the present
situation of the primate importation and make practical guidelines for
preventing zoonoses caused by imported primates.
The following issues were discussed: (I.) present state of the
importation of primates and the countries of habitat, (II) measures for safe
transportation, (III) management and handling on arrival at an air port, (IV)
quarantine, (V) items and
application of quarantine examination, and (VI) input and output of information.
Page-26 Announcement from the CPRLP
Now,
the CPRLP is recruiting supporting members.
Page-27
Introduction of facilities: Information center of the joint use facility for
research
with monkeys
It was decided that an information center will be constructed from this
June. The center will be attached
to the joint use facility for research with monkeys, of which we have reported
in the previous issue ( See, TPC News Vol. 15, No.2).
The floor plan of the first floor is shown at the bottom of the page.@
The information center involves three information rooms, one meeting room
and an accommodation area for visiting researchers.
This center is expected to be useful for yielding further utilization of
the joint use facility for research with monkeys.
Page-28
Comment from a trainee: Two
phantoms
Mr.
Usami, who had been a trainee at the TPC for three years, outlined his result,
with a lot of thanks to the TPCfs people, including Dr. Yoshida, his
supervisor.
Mr.
Usami came to the TPC to establish the animal model of osteopenia in the African
green monkey. He and Dr. Yoshida
had to design new calibration instruments shaped phantom to measure more
correctly the mineral density of the spinal bone and whole body with a dual
energy X-ray absorptiometry (DXA).
The
two calibration phantoms are shown in the figures.
Page-29
Report on the Seminar f97 by the CPRLP
The third seminar by the CPRLP was held on September 14, 1997, at the
Tsukuba Center for Research in Tsukuba city.
Mr. Hiyaoka, who was a secretary of the Seminar, reports the outline of
the Seminar.
The theme of this year was g
Anesthesia for laboratory primates: foundation and practice .h
Four sessions by six lecturers: 1) clinical practice in humans, 2)
clinical practice in animals, 3) effect on experiments and its countermeasure,
and 4) techniques in handling monkeys without anesthesia, resulted in hot
questions and answers among the participants after every lecture.
Mr.
Hiyaoka concludes that the seminar was highly successful.
Page-30
Report on g 14th Symposium on Non-human Primate Model for AIDSh
Dr.
Mori has been to Portland, USA, last year to attended g 14th
Symposium on Non-human Primate Model for AIDS g, which was held on October 23-
26, under the auspices of the Oregon Regional Primate Research Center (ORPRC),
Beaverton, Oregon.
Prior
to attending the symposium he visited the ORPRC.
There, he was very impressed of the advanced facilities for animal
experimentation as well as biosafety
level containment facilities.
He outlines some sessions he was interested in, including three reports
from Aaron Diamond AIDS Research Center, by Drs. David Ho, Preston Marx and Zwei
Chen.
Page-31
From the Primate Forum
At the present, three serial online-lectures are running in the Primate
Forum (PF).
gZoonosesh by Dr.
Yamanouchi, gResearch on growth and laboratory animalsh and
gLetter from Uruguayh by Dr. Gotoh, and gPopulation geneticsh by
Dr. Yasuda.
Every
title of the lectures uploaded into the PF so far were listed up.