14 research outputs found
Relationship between the histological structures of spleen and its possible functions in the immune response
The spleens of human, ox, rabbits and rats were
studied histologically. Serial sections of the tissues
stained with methyl-green pyronin, Mallory-azan
and Gridleys silver method were examined carefully
in regarding to its architecture and staining nature
of the composed cells.
Pyronin-positive plasma cells were found to be
localized at the periarteriallymphocyte sheath which
surrounds the central arteries. The cells were more
numerous at the terminal portion of the central
arteries.
Two distinct layers, the outer and the inner, were
recognized and the apposed borders of each layer
were separated by the reticular membrane which
contains numerous marginal sinuses and capillaries.
Composing cells of the marginal zone characterized
by the small amount of pyroninophi1ic cytoplasm
and lighter nucleus were different from the small
lymphocytes and the macrophages.
A nu ;nber of macrophages and rich capillaries were
observed in the outer layer of the marginal zone.
The so-called dark zone and the light zone of the
g섬rminal center seemed to have constant direction of
the arrangement, that is. the dark zone was located
always near the central artery, while the light zone
was far from the central artery.
The possible invading route of the antigen in
relation to the structures were discussed
Effects· of the Azathioprine Administered during Pregnancy on the Lymphoid Orga n s of the Neonatal Rats.
The following experiment was performed to study
the effect of azathioprine administered during pregn'
ancy on the development of lymphoid organs of neo'
natal rats.
The experimental animals were the three hundred
Sprague·Dawley neonatal rats which were divided into
four groups. The animals born from the azathioprine'
administered adults during pregnancy were named
for group 1 and 2. and were injected with antigen
to group 1 and saline to group 2 respectively after
birth. The animals of group 3 and 4 were born from
the adults free of azathioprine during pregnancy and
were injected with antigen to group 3 and saline to
group 4 respectively after birth.
Antigen or saline was injected intraperitoneally on
the 1st. 4th. 9th. 12th and 16th week after birth.
Then the spleen and thymus were resected on the
1st. 3rd. and 7th day after the injection for the plaque
assay and histochemical studies
The following results were obtained:
1. No plaque·forming cells and large pyroninophilic
cells were observed in the spleens of all groups on
the first week after birth
2. The number of plaque·forming cells and large
pyroninoph i1ic cells observed in the spleens was much
less after antigen injection but a little less after
saline injection in the group of azathioprine admini
stration than in the control group on the 4th. 8th
and 12th week after birth.
3. A similar number of many plaque·forming cells
and large pyroninophi1ic cells were observed in the all
groups on the 16th week after birth.
4. While the development of thymus was retMded
histologically for the groups of azathioprme acimini·
stration than the control groups until the· 8th week
after birth. a similar development of thymus between
the two groups was observed on the 12th and 16h
week after birth.
5. It may be suggested that adminislration of
azathioprine did interfere with the development of
T·lymphocyte and censequently affected the formation
of antibody due to the insu뻐cient collaboration between
T cells and B cells
Metabolism of C14-labeled Pyruvate and Glucose by Brain-Slices of Normal and Alloxan-Diabetic Rats.
Brain slices of 58 normal and alloxan-diabetic rats
were divided into 4 groups. Each group was incubated
for a period of 5 hours with incubation mixture
consisting of 15 C.c. of phosphate buffer to which
were added C1L glucose or C14-pyruvatc and 15 C.c
of oxygen. During 5 hours of incubation period. the
incubation media and gas phase were replaced with
fresh media and oxygen at hourly intervals. This
procedure made it possible to obtain hourly samples to determine the concentration of medium substrates
and the amount and radioactivity of CO2 produced by
the brain slices during the experimental period. It
also had the advantage of presenting the tissue with
a substrate of essentially constant concentration and
specific activity.
Data obtained from the C14-pyruvate incubation
experiment, in the first and second group , and from
the ClL glucose incubation experiment in the third and
fourth group , were summarized as the follows.
1. In the ClL pyruvate incubation experiment, pyruvate
disappearance rates were mean of 10.7 μM/hr/g
in the first control group , and 12. 2 μM/hr/g in
the second alloxan-diabetic group. The lactate accumulation
rates in both groups were mean of 5. 8 and
5.1 μM/hr/g respectively.
Total CO2 production rates in both groups were
mean of 49. 7 and 63. 4 μM/hr/g respectively, showing
a slightly higher value in the second group.
However, since RSA, which represents the fraction
of C14-02 derived from medium CILpyruvate to total
CO2 production rate, was slightly hiher in the control
group than the diabetic-group, there were little diffe•
rences in C02 production rates derived from medium
CILpyruvate of both groups.
2. In the C14-glucose incubation experiment, ClL
glucose disappearance rates decreased about one half
of the control in the diabetic group , showing a mean
of 44.1 μM/hr/g in the third control group and
20.0 μM/hr/g in the fourth alloxan-diabetic group.
Since there were not significant differences in total
CO2 production rates and values in RSA, Cl4-0 2 production
rates derived from medium clιglucose have
been found to be almost the same value in both
groups.
From the data described above, insulin cannot have
an influence on all the steps of carbohydrate metab
olism in the brain tissue. However there is evidence
that insulin exhibits the effect on membrane transport
of glucose as found in the other tissues
Experimental Microvascular Grafts in Rats and Its Histological Studies.
Histopathological features of microvascular anasto!
TIGses using "Invagination technique," end-to-end anastomoses and long term results of interposed venous
grafts were studied in the femoral vessels of rat
one to 12 months following surgery. The observation
performed under light microscope, TEM and SEM.
I. Histopathological features of the anastomotic site,
which were repaired by "Invagination technique"
and usual end·to·end anastomosis respectively,
did not differ each other in the long term results.
2. Histological regeneration of anastomotic site did
not accomplished by post·op 2 months and revealed
active process was undergoing.
3. Endothelial lining of intima was nearly completed
by 2 months at undamaged portion directly by
suture materials.
4. In summary it may be said that histological
examination of the specimen ranging in age from
one to 12 months led general conclusion that
venous grafts persist as living structure but
undergo certain histological changes consisting of
fibrous reinforcement
Production of Monoclonal Antibody for Vasopressin
Monoclonal antibodies were produced against arginine
vasopressin (AVP) by fusion of mouse myeloma
cells with hyperimmune mouse spleen cells. Modified
radioimmunoassay(RIA) was used to detect the
presence of specific antibody in culture. Specificity
of the antibody was further characterized by enzyme
linked immunospecific assay(ELISA). This antibody
was able to localize differentially the vasopressin
neuron from the oxytocin neuron in tissue sections
of the hypothalamus
Histochemical Changes of the Rat Lymphatic Organs in Relation with the Altered Status of Thyroxin
To observe the effects of thyroxine on the lymphatic
organs, the Sprague-Dawley rats weighing about
200g were classified into 4 groups (Group 1: control,
Group 2: thyroidectomy, Group 3: tyroidectomy plus
thyroxin treatment in the dose of restoring the metabolism
to normal, Group 4: thyroxin treatment in
the dose of inducing hyperthyroidism), and from
each experimental animals thymus, lymph node and
spleen were resected on the Ist, 3rd, 5th and 8th
week and fixed in 10% neutral formalin after measuring
the organ weight.
Each sections were stained with Hematoxylin-Eosin,
Methyl Green Pyronin, PAS and Silver stain and
observed under the light microscopy.
The results are summarized as follows.
1. Weights of the lymphatic organs per body
weight decreased after thyroidectomy and increased
after thyroxin treatment.
2. After thyroxin treatment:
1) Pyronin positive lymphocytes increased in the
corticomedullary junction of thymus, germinal
center of the lymph node and periarteriallympho
cyte sheath of the spleen.
2) PAS positive cells increased in the corticomedullary
junction of thymus.
3) Reticular fibers increased in the medulla of thy
mus and perifollicular region of the lymph node.
4) Hematopoietic cells increased in the red pulp of
spleen.
3. After thyroidectomy the density of chromatin
in the nucleus of thymocytes decreased, and the
lymphoid follicles decreased in size and number. 4. The results suggested that thyroxin acts as a
mitogen and effects the proliferation and differentiation
of lymphocytes
Study on the Inhibition of the Immune Response of the Neonatal Rat Spleen by the Azathioprine Administered during Pregnancy
The following experiment was performed to study the inhibition mechanism of the immune response in
the neonatal rat spleen by azathioprine administered
during pregnancy.
The experimental animals were the two hundred
neonatal rats which were born by sixty normal Sqrague-
Dawley pregnant rats, of which 45 rats 8mgjkg
of azathioprine were administered to orally at the
7th gestational day.
Above two hundred experimental animals were
divided into 6 groups as follows:
Group 1: Neonatal rats to which T-dependent antigen,
Sheep Red Blood Cells (SRBC)
were injected intraperitoneally, born by
normal pregnant rats.
Group 2: Neonatal rats to which SRBC were injected
intraperitoneally, born by pregnant
rats to which azathioprine had already
been given orally during pregnancy.
Group 3: Neonatal rats to which SRBC were injected
intraperitoneally and thymus cells
derived from outbred neonatal rats were
injected intravenously, born by pregnant
rats to which azathioprine had already
been given orally during pregnancy.
Group 4: Neonatal rats to which T-independent
antigen, Dextran was injected intraperitoneally,
born by normal pregnant rats.
Group 5: Neonatal rats to which Dextran was
injected intraperitoneally, born by pregnant
rats to which azathioprine had already
been given orally during pregnancy.
Group 6: Neonatal rats to which Dextran was
injected intraperitoneally and thymus
cells derived from outbrcd neonatal rats
were injected intravenously, born by
pregnant rats to which azathioprine had
already been given orally during pregnancy.
Antigen was injected intraperitoneally to above
each group on the 1th, 8th and 12th weeks after
birth and then, the spleen was resected on the 3rd
and 7th days after injection of antigen for the plaque
assay and histochemical studies.
The following results were obtained:
1. Many plaque-forming cellsCPFC) were observed on the 3rd day after injection of antigen in all
groups to which SRBC were injected, while many
PFC were observed on the 7th day after injection
of antigen in all groups to which Dextran was
injected.
2. More PFC were observed in Group 1 than Group
2, and much more PFC were observed in Group
3.
3. There were no significant differences in number
of PFC between Dextran·Groups(Group 4, Group
5 and Group 6).
4. There was strong tendency to recover the immunoIogicaI
function with increasing age in SRBC·
Groups(Group 1, Group 2 and Group 3).
5. Above results were in good accordance with
those of histochemical observations.
6. It is strongly suggested that administration of
azathioprine during pregnancy did suppress the
development of fetal thymus, and that as a result
of that suppression peripheral Tvhelper cells were
depleted and consequently the formation of antibody
was inhibited due to the insufficient colla'
boration between T cells and B cells
Immunological and Histological Studies on the GALT and the Other Lymphoid Organs in the Rabbits
As an experiment for the study of bursa equivaleot
organ in mammalia, the present experiment was
performed on the GALT of young adult rabbits
whether the cells of it respond to the antigenic
stimulation or not.
The experimental animals, weighed 1. 5 to 1.8kg,
were randomly divided into two groups. Experimental
group received a single injection of 1 ml of 20%
sheep erythrocytes intravenously. Control group
received a single injection of 1 ml of saline solut ion
intravenously. After the injection of the antigen
(or saline) the animals at various interval (from 1st
day to 21st day) and the palatine tonsil, Peyer's
patch, appendix, spleen , and thymus were removed.
The cell suspensions obtained from each organ was
used for the plaque technique and the paraffin
sections obtained from each organ were carried
out for the staining with methyl-green pyronin,
silver, and hematoxylin-eosin. Hemolysin-producing
cells were determined using a modification of Jerne
plaque technique and the results were expressed as
plaque-forming cells per 10' cells.
The results were as followings:
1. In the experimental group the spleen and
palatine tonsil produced a number of plaqueforming
cells. It reached to the maximum at 3
days after the injection of the antigen and
then decreased gradually.
2. By contrast, thymus, appendix, and PeyerS
patch did not produce the plaque•forming cells
as did the spleen and palatine tonsil.
3. The number of large pyroninophilic cells were
markedly increased in the extrafollicular area
of palatine tonsil and the periarteriallymphocyte
sheath of spleen after the antigenic stimulation.
It reached to the maximum at 3 days after the
injection of antigen and then decreased
gradually.
4. Number of large pyroninophilic cells did not
change in the appendix, Peyers patch, and
thymus after the antigenic stimulation.
5. The diffuse lymphatic tissue of the extrafolli
cular area of palatine tonsil were more well
developed than that of appendix and PeyerS
patch.
These results suggest that the palatine tonsil in
the rabbits as well as spleen responds to the
antigenic stirn ulation, while the appendix and
Peyers patch do not
Studies on the Plasma Cells in the Lymphoid Organ during Pregnancy and Lactation in the Mouse I. Observations in the Thymus
The following experiment was performed to study
on the numerical changes of the IgG, IgM and IgA
bearing cells in the thymus during pregnancy and
lactation in the mouse. The experimental animals
were mated allogeneically (female ICRC x male
DDY), and the thymi were resected at 4, 6 and 19
days of gestation and postpartum 20 days. The thymi
were stained with MGP and immunohistochemical
reaction. Then each group was compared with the
virgin control group.
The following results were obtained:
1. In 6 days of gestation, the number of IgA bearing
cells was significantly more than that in virgin.
2. In 19 days of gestation, the number of IgG. IgM
and IgA bearing cells was significantly more
than that in virgin.
3. In suckling group, the number of IgG and IgA
bearing cells was significantly more than that
in virgin.
4. The immunoglobulin bearing cells were localized
in perivascular region.
On the basis of the above results, it is suggested
that the thymus. the primary lymphoid organ, has
relation with the maternal immune response, too
A Case of Anencephalus
A case of anencephalus. 38 weeks in fetal age. was
observed.
1) The calvaria was completely absent and whole
of the vertebrae showed complete spina bifida. There
was strong lordosis between the 7th cervical and lst
thoracic vertebra. which were compressed under the
skull base.
2) Brain was completely absent while the bifurcated
spinal cord was exposured on its upper half. Cranial
nerves. of which peripheral part were existed. end in
neighboring soft tissue or anastomosed with cervical
nerves.
3) Marked enlargement or the liver was observed.
4) Absence of hypophysis and remarkable atrophy
of the adrenal glands. characteristic features of the
anencephalus. were observed in this case.
5) Profuse amount of amnionic fluid was detected
in delivery.A case of anencephalus. 38 weeks in fetal age. was
observed.
1) The calvaria was completely absent and whole
of the vertebrae showed complete spina bifida. There
was strong lordosis between the 7th cervical and lst
thoracic vertebra. which were compressed under the
skull base.
2) Brain was completely absent while the bifurcated
spinal cord was exposured on its upper half. Cranial
nerves. of which peripheral part were existed. end in
neighboring soft tissue or anastomosed with cervical
nerves.
3) Marked enlargement or the liver was observed.
4) Absence of hypophysis and remarkable atrophy
of the adrenal glands. characteristic features of the
anencephalus. were observed in this case.
5) Profuse amount of amnionic fluid was detected
in delivery