The generation of large pyroninophilic cells in the lymphoid tissues of rats infused with cell-free lymph.

The thoracic duct of Wistar strain rats was cannulated during 5 days for studying the effect of selective lymphocyte depletion on the lymphoid tissue. A technique for the continuous infusion of cell-free lymph, whole lymph of Eagle's medium to the rat with the thoracic duct fistula is described in detail. The prolonged drainage of lymph from rats was followed by lymphopenia, sever atrophy of lymphoid tissues and the depletion of small lymphocytes in the thymus-dependent areas of spleen and lymph nodes. The infusion of cell-free lymph into the drained rat resulted in the recovery of the weight of lymphoid tissues and in the massive proliferation and accumulation of large cells with prominent nucleoli and intensely pyroninophilic cytoplasm in the lymphocyte depleted areas of the peripheral lymphoid tissues and thymic cortex. There was histological evidence that the large pyroninophilic cells developed well in the spleen and tended to localize preferentially around the periarteriolar region through the marginal zone bridging channels to the red pulp. The infusion of Eagle's medium was found ineffective in restoring the weight of the lymphoid tissues and in bringing about the proliferation of lymphoid cells. The rats infused with whole lymph showed almost similar findings biologically and histologically to those of sham-operated rats.     

Repair processes of hemopoiesis after applying cyclophosphamide. II. Quantitative changes in basic proteins and DNA in the bone marrow, spleen, and thymus

The effect of cyclophosphamide on the organ weight, DNA and basic protein content in nuclei of bone marrow, thymus and spleen of rats was studied. Animals were examined 6, 12 and 24 hours and 3, 5, 10, 20 and 30 days after intraperitoneal application of cyclophosphamide in the doses of 100 and 200 mg X kg-1 of body weight. From the 5th day after application a marked decrease in weight and cellularity of all organs was observed. Amounts of DNA and basic proteins in nuclei of bone marrow and spleen were higher as compared with controls and their increase occurred prior to recovery of cellularity and organ weight, that was initiated within 5 and 10 days after cyclophosphamide treatment. Changes in the thymus persisted for a longer period of time and recovery was incomplete even at the day 30 after cyclophosphamide application. In accordance with these data no increase in DNA and basic protein amounts in the thymus nuclei was observed.     

Sensitivity and specificity of the bioassay of estrogenicity in mammary gland and seminal vesicles of male mice

Young intact (18 days old) and adult castrated males of CBA and C3H/Di mice were used for measuring the estrogenicity on the basis of growth response of mammary epithelial structures and the weight of seminal vesicles. It was demonstrated that heavier young males had disproportionally heavier seminal vesicles (sex steroid-responsive organs) than small animals at day 33 of age (that is on the day when experimental animals were killed and organs dissected). However, the weight of the spleen (sex steroid-nonresponsive organ) was proportionally related to body weight. To minimize variability in hormone responsiveness, all animals were weighed at the age of 18 days and only males weighing 8+/-1 g were used for hormone treatment. The percentage area of mammary fat pad occupiedby mammary epithelial structures was progressively increased by 17beta estradiol from dose 0.01 microg x d(-1). The maximum effective dose of estradiol was 0.1 microg x d(-1) and dose 10 microg x d(-1) of estradiol decreased mammary size to control level (inverted-U-shaped dose-response curve). Progesterone alone stimulated mammary growth only in high doses (500 microg x d(-1) and higher) in young intact males, but had no effect on mammary growth in adult castrated animals. In young intact males, estradiol alone, or progesterone alone decreased the weight of seminal vesicles. No such inhibitory effect of these hormones was noted in adult castrated males. Progesterone acted synergistically with estradiol to produce higher mammary growth compared to that in males treated with estradiol alone. In the presence of progesterone seminal vesicles weight was decreased by estradiol given in such low doses as 0.001 microg x d(-1) of estradiol, which is 10 times lower than that effective in animals treated with estradiol alone. On the other hand, in the adult castrated males a combination of estradiol plus progesterone stimulated seminal vesicles weight. The effects of a combination of estradiol plus progesterone in the mammary gland were mimicked by norethindrone acetate (a synthetic steroid exhibiting progestantial and estrogenic activities) and inhibited by both testosterone and cortisol. Estradiol, progesterone, norethindrone acetate, or testosterone did not affect spleen weight and size of mammary lymph nodes.However, cortisol significantly decreased not only spleen weights but also size of mammary lymph nodes. These results showthat simultaneous evaluation of mammary gland growth, seminal vesicles, and the spleen weight in the same animal is suitable for bioassay of estrogenicity as well as for detection of androgenic and antiandrogenic activities.     

Short- and long-term effects of whole-body irradiation with fission neutrons or X rays on the thymus in CBA mice

Young adult (6 weeks old) female CBA mice were exposed to whole-body irradiation with either 2.5-Gy fast fission neutrons of 1 MeV mean energy or 6.0-Gy 300 kVp X rays at centerline dose rates of 0.1 and 0.3 Gy/min, respectively. The weight of spleen and animal and the weight, cellularity, and histological structure of the thymus were studied at different times after irradiation. Thymic recovery after whole-body irradiation showed a biphasic pattern with minima at 5 and 21 days after irradiation and peaks of regeneration at Days 14 and 42 after X irradiation or at Days 14 and 70 after neutron irradiation. After the second phase of recovery, a marked decrease in relative thymus weight and cellularity was observed, which lasted up to at least 250 days after irradiation. Splenic recovery showed a monophasic pattern with an overshoot on Day 21 after irradiation. After neutron irradiation a late decrease in relative spleen and animal weight was observed. The observed late effects on thymus and spleen weight and thymus cellularity are discussed in terms of a persistent defect in the bone marrow.     

The effect of corticosteroids upon the number and organ distribution of "background" immunoglobulin-secreting cells in mice

The influence of the synthetic corticosteroid dexamethasone sodium phosphate (DEXA) upon the immunoglobulin (Ig)-secreting cells was studied in not intentionally immunized BALB/c mice. This was done for IgM-, IgG-, and IgA-secreting cells in spleen, mesenteric lymph nodes (MLN), and bone marrow (BM). A single injection of DEXA (16 to 144 mg/kg body wt) markedly reduced the number of Ig-secreting cells in spleen and MLN within 1 day, but hardly affected their number in the BM. The decrease was immediately followed by a recovery and, at the highest doses and especially in MLN, by an overshoot. Two weeks after the initial decrease a second decrease was found. When mice were subjected to daily treatment with DEXA during 1 week, initially a recovery pattern was found in spleen and MLN similar to that found after a single injection of a high dose. In this case, however, the effects were less dose dependent, and the overshoot reaction was followed by a period of subnormal numbers of Ig-secreting cells which lasted at least 1 week. This late effect of DEXA not only occurred in spleen and MLN, but also in the BM. The most prominent effect of daily treatment with DEXA was the long-lasting decrease of the number of IgG-secreting cells starting 1 week after withdrawal of treatment. This decrease was associated with a severely decreased serum IgG level.     

Changed murine lymphocyte trapping after treatment with Propionibacterium granulosum

Summary The trapping of lymphocytes occurring at different times after a single injection of Propionibacterium granulosum was studied. Labeled syngeneic lymphocytes injected into Propionibacterium-treated recipients showed a different pattern of localization from that observed in untreated animals. A pronounced decrease in homing to the lymph nodes and spleen and an increase in localization in the liver were found. The extent of trapping in the liver corresponded to the increase in weight of this organ. Whole-body irradiation with 400 R, used to achieve autologic lymphocyte depletion, did not change the localization of labeled cells. However, macrophage damage caused by silica resulted in diminished trapping in the liver in Propionibacterium-treated animals and increased accumulation of labeled lymphocytes in the spleen and lymph nodes.     

Action of Co60 gamma radiation on the nucleic acid concentration and ribonuclease activity in the splenic tissues of inbred and hybrid mice]

The spleen tissue radiation injury expressed in the organ weight loss, nucleic acid concentration decrease and ribonuclease activity increase was observed to a greater extent in mice of the AKR line and to a less extent in those of C57BL line; C57BL X AKR hybrids occupied an intermediate position. It shows that animal radiosensitivity is probably determined by the genotype.     

The effect of feeding 11-ketotestosterone on the food conversion efficiency and tissue protein and nucleic acid contents of juvenile carp, Cyprinus carpio L.

Juvenile common carp were fed 11-ketotestosterone for 60 days with the diet and the effect on food conversion efficiency, organ weights and protein and nucleic acids (RNA, DNA) content of the liver, kidney, brain and muscle were observed. Feeding of the steroid increased the food conversion efficiency of all the experimental groups studied as compared with controls. A decrease in weight of brain, liver and kidney in relation to body weight was noticed after 60 days of the hormone feeding. No change in the visceral weight was observed. These changes in relative weights of the organ were ameliorated 30 days after the withdrawal of the steroid from the food. The weight of the viscera decreased compared with the control weight during this time. Feeding of the hormone brought variable changes in the total proteins, RNA/DNA, protein/RNA and protein/DNA in all the organs studied. In addition to these findings, changes in the moisture, total lipid and ash contents of the muscle were also observed. The results are discussed in the light of existing knowledge of the effect of anabolic-androgenic steroids on the growth processes of different organisms.     

The effect of primary and secondary immunization on the lymphoid tissues of the carp, Cyprinus carpio L

Mirror carp immunized with human gamma globulin (HGG) in Freund's complete adjuvant (FCA) show a proliferative response involving cells whose cytoplasm stains deep red with methyl green-pyronin (pyroninophilic cells). This response occurs particularly in the haemopoietic parenchyma of the pronephros and mesonephros. It peaks at week 3, with the formation of clusters of pyroninophilic cells in the pronephros. Immunization with Aeromonas salmonicida elicited a less intense pyroninophilic response but caused a larger increase in pigment-containing cells. After a secondary immunization with HGG in FCA, a distinct response was observed in the spleen: Pyroninophilic cells collected within the ellipsoid sheaths in large numbers and formed nodules. The reticulum of such nodules acquired spherical proportions and resembled the white pulp reticulum of the tetrapod spleen. The roles of such pyroninophilic cells and the possibility that aggregations of them may be functionally analogous to homoiotherm germinal centres are discussed.     

Lymphoid organ weights and organ:body weight ratios of growing beagles

Although dogs, especially beagles, are used extensively in biological and clinical investigations, the literature dealing with normal biological measurements of their lymphoid organs is scanty. This study was undertaken to provide the information on the weight of lymphoid organs of beagles. The thymus, spleen, and prescapular, popliteal, and mesenteric lymph nodes of 95 normal beagle dogs, from one day to 11 months of age, were weighed and compared with body weights. The weight of the thymus and spleen increased drastically at and after 2 months of age, although the organ:body weight ratios remained the same at 2 months of age and decreased afterward. Similar increases in the weight of the mesenteric lymph node complex, but with an increase in the organ:body weight ratio, occurred also at and after 2 months of age, reflecting the importance of the gut-associated lymphoid organs after weaning. The increases in the size of the cutaneous nodes, prescapular and popliteal, were less pronounced and their organ:body weight ratios remained the same from birth through 11 months of age.     

Changes in spleen histology in response to antigenic stimulation in the snapping turtle,Chelydra serpentine

Proliferative and migratory changes of lymphoid cells in the spleen were observed in turtles stimulated with KLH and maintained at 30°C. Small foci of pyroninophilic lymphoblasts were first seen in the white pulp at about day 5. Progressive enlargement of these centers continued and peaked by days 8-12. By days 15-20 the white pulp returned to a normal (unimmunized) state, while the number of pyroninophilic cells, primarily plasma cells, increased markedly in the red pulp. At days 22-25, the number of plasma cells returned to normal levels and the spleen appeared normal for the remainder of the 60 day observation period. These events suggest that at 30°C,-the turtle is capable of a strong and prompt proliferative response in the white pulp sheaths, followed by migration and differentiation of lymphoblasts into plasma cells, n the red pulp., Observations of pyroninophilic cells in sinuses, venules and veins of the spleen and a concomitant depletion of cells in red pulp, further suggest a migration from the spleen to other sites. Following a second antigenic challenge, at day 60, no significant histological changes were observed at 30°C. Nor were any changes observed following primary or secondary antigenic challenge, in animals maintained at 10°C. These findings are discussed with regard to immunological memory and low temperature immunosuppression in ectothermic vertebrates.     

3H-deoxythymidine incorporation in graft-versus-host disease in the Norway rat. I. Liquid scintillation studies.

Graft-versus-host-disease was produced in newborn Brown Norway (BN) rats with an intravenous (iv) injection of adult allogeneic Lewis (L) lymph node cells (experimental) and the response was compared to littermates injected with adult syngeneic BN cells (control). By 4 days the reaction in the spleen of experimental animals was such that the spleen index was 1.70 and 2.58 on day 7, and continued to increase until death. A one hour iv pulse of tritiated deoxythymidine (3HdT) administered to experimental and control animals revealed a whole organ peak incorporation of 3HdT on day 6 in experimental spleens. A second larger peak occurred on day 10 in the experimental spleen as compared to a single peak at days 6 or 7, respectively, in the experimental mesenteric and combined superficial lymph nodes. However, analysis of the incorporation of 3HdT with respect to organ weight revealed a peak incorporation in animals receiving L cells on day 4--6 with a second smaller peak on day 10 in the experimental spleen and again a single peak on day 5 or 6 in the lymph nodes. Total 3HdT incorporation within both experimental lymph node compartments became less than controls by day 15 even though experimental nodes had a larger mass. 3HdT incorporation per milligram tissue weight decreased in all tissue compartments of experimental animals by day 13--14. The contribution of donor and host cell proliferation to the various peaks observed is discussed.     

COMPARISON OF DNA RENEWAL IN GERM-FREE AND CONVENTIONAL MICE USING [125I]IODODEOXYURIDINE AND [3H]THYMIDINE

Germ-free (GF) and conventional (CV) C3H mice received a single injection of 1 μCi [³H]thymidine and 3 μCi [¹²⁵I]iododeoxyuridine to provide simultaneous labeling of DNA with the two precursors. Thymus, spleen, mesenteric lymph nodes, bone marrow (femora), small intestine, colon and skin were examined for total organ activity and rate of DNA renewal 1–8 days after injection. Precursor incorporation, assayed on day 1, was lower in the thymus, mesenteric lymph nodes and femora (and, to a lesser extent, in the spleen and colon) of GF mice as compared to CV animals. The opposite was observed in the small intestine and skin, i.e. total organ activity was higher in GF animals. Differences in precursor incorporation were partly due to differences in organ weights between the two groups of mice. In comparison to CV animals, DNA renewal rates were diminished in the mesenteric lymph nodes, bone marrow, colon (following a 3-day plateau) and spleen of GF mice. Little, if any, difference was observed between the two groups with respect to the rate of DNA turnover in the thymus and skin. Radioactivity of the small intestine remained constant for 2 days. Thereafter intestinal activity in GF mice declined at an initial slow rate between days 2 and 5 followed by a rapid decrease between days 5 and 8. In CV mice the first phase of activity loss was short with the rapid decline in intestinal activity beginning on day 3. From the slopes of the regression lines, the percentage thymidine reutilization was estimated. Reutilization varied from 0 to 63% in the various organs examined, with the greatest difference between GF and CV mice occurring in the mesenteric lymph nodes.     

不同周龄SD大鼠脏器重量及其变化趋势分析

目的分析不同周龄SD大鼠的脏器重量及其变化趋势,为评判药物毒性反应提供理论参考。方法分别选取试验第13、26、52、78和104周对照组动物脑、脾脏、心脏、肺脏、肝脏、肾脏、肾上腺、睾丸、卵巢的重量数据并分析。结果从13～104周SD雌鼠脑、脾脏、心脏、肺脏、肝脏、肾脏、肾上腺、卵巢的重量呈升高趋势。从13～104周SD雄鼠脑、脾脏、心脏、肺脏、肝脏、肾脏重量均重于雌鼠,但雌鼠肾上腺重量、脏体比和脏脑比均显著高于雄鼠。结论本研究首次在国内建立了符合我国实验动物现状的,不同周龄SD大鼠的脏器重量背景数据和参考值范围,并分析了不同周龄SD大鼠脏器重量变化趋势。     

Radiation sensitivity of hemopoietic stroma: long-term partial recovery of hemopoietic stromal damage in mice treated during growth

We studied the ability of the hemopoietic organ stroma to recover from damage inflicted by 5 or 7 Gy gamma radiation administered during a period of stromal growth in 4-week-old mice. Irradiation resulted in an immediate depletion of femoral colony-forming fibroblastic progenitors (CFU-F) down to 10-20% of age-matched control values. A full recovery to normal numbers occurred between 120 and 240 days after irradiation and was followed by a secondary decrease 1 year after irradiation. This secondary decrease was accompanied by a decrease in the femoral CFU-S and CFU-C content. Femoral CFU-F attained normal numbers and it was demonstrated to occur from surviving CFU-F and could not be enhanced or prolonged following infusion of unirradiated bone marrow cells after irradiation. During the transient CFU-F recovery the hemopoietic stroma remained severely damaged as judged by the regenerative capacity of spleen and femur stroma after subcutaneous implantation, and the ability of the spleen to accumulate CFU-S in response to lipopolysaccharide injection. We have reported earlier that in similarly irradiated adult mice, no restoration of femoral CFU-F was observed. This difference between 4-week-old and adult mice could not be explained by a difference in in vitro radiosensitivity of CFU-F or in their in vivo regeneration kinetics following irradiation and subsequent lipopolysaccharide injection. We conclude from these observations that the recovery kinetics of the CFU-F population is different in young and adult irradiated mice, infused CFU-F do not contribute to CFU-F regeneration in an irradiated femur, CFU-F are not the sole determinants of stromal regeneration in femur and spleen following irradiation.     

Histological changes in spleen and lymph nodes of mice administered cyclophosphamide and levan

Summary Morphological changes in spleen and lymph nodes of C57B1 mice induced by the cytotoxic agent cyclophosphamide and the polysaccharide levan, separately and in combination were studied. In the spleen, an early decrease (phase 1) and a late increase (phase 2) in weight were found to result from all drug administrations. The hypocellularity of phase 1 was due to a depletion in the white pulp affecting mainly the B-region. Splenic weight decrease was parallel to B-cell depletion and most marked in animals exposed to cyclophosphamide with levan. The splenomegaly observed during phase 2 with all treatments was due to extramedullary hematopoiesis in the red pulp. In the lymph nodes, administration of cyclophosphamide and levan produced opposite effects on the B-cell region: cyclophosphamide eliminated the germinal centers for 8 days, but levan seemed to enhance B-cell activity. In animals given both cyclophosphamide and levan, inhibition of B-cell activity began earlier than with cyclophosphamide alone. Levan does not antagonize the suppressive effect of cyclophosphamide on the B-cell component at the early phase when the drugs are given together.     

Biosynthetic human growth hormone: effects on growth of Snell dwarf mice

Bacterially synthesized human growth hormone (bhGH) administered to Snell dwarf mice during 4 weeks, induced an increase in body length and weight to a comparable degree as obtained with pituitary-derived human growth hormone (hGH). At a dose of 150 mU/day both bhGH and hGH induced a significant stimulation over saline-treated controls, of the weight of the submandibular salivary glands, the m. quadriceps femoris and gastrocnemius, the heart, liver, kidneys, thymus and spleen. The weight of the brain and the thickness of the skinfold were not influenced by either of the preparations used. When organ weights were expressed as a function of body weight, the contribution of the kidneys to body weight was significantly higher with hGH than with bhGH. The other organs studied did not show differences. As a biochemical parameter of cartilage growth, the sulfate incorporation into costal and epiphyseal cartilage in vitro was measured, and it was found to be stimulated by both hormones after short-term treatment. Thus bacterially synthesized hGH behaves identically to pituitary-derived hGH with respect to body length, sulfate incorporation into costal and epiphyseal cartilage, body weight and organ growth of Snell dwarf mice, with one exception: increase of weight of the kidneys, as a function of body weight, was more pronounced after treatment with hGH than with bhGH.     

Dynamics of postnatal histogenesis of the thyroid in normal rats and after sympathectomy

The changes of the thyroid gland and neurocytes of the cranial sympathetic ganglia were followed in rats of different ages after guanethidine injections with the use of radioimmunological assay, electron microscopy and morphometry. The injection of 15 mg of the drug per kg of body weight within the first two weeks after birth caused the death of over 80% of the cells in the sympathetic ganglion. In the sympathectomized 15-day- and 1-month-old rats the functional activity of the thyroid gland was markedly reduced. Later on, intrathyroid hormonogenesis somewhat increases due, apparently, to partial recovery of the organ adrenergic innervation and increase in the production of thyrotropic hypophysial hormone and calcitonin.     

B-Lymphocyte differentiation in lethally irradiated and reconstituted mice. II. Recovery of humoral immune responsiveness.

The recovery of humoral immune responsiveness was studied in lethally irradiated, fetal liver-reconstituted mice. By means of both membrane fluorescence and antibody formation to sheep red blood cells (SRBC) as a functional assay, the rate of recovery of the compartments of B and T lymphocytes was determined in various lymphoid organs. The recovery of the immunoglobulin-positive (B) cell compartment after irradiation and reconstitution started in the spleen. This organ was also found to be the first in which the recovery of the B-cell population was completed. The interval between the recovery of the B-cell population in the spleen and that in the other organs tested was found to increase when the irradiated mice were reconstituted with spleen colony cells instead of fetal liver cells. This proved to be caused by the number and nature of the reconstituting hemopoietic stem cells. The immunoglobulin-positive (B) cells were found to appear before SRBC-reactive B cells could be demonstrated in spleen, lymph nodes, and Peyer's patches. The appearance of T lymphocytes in the various lymphoid organs required even more time. By means of cell transfer experiments, a sequential appearance of the precursors of anti-SRBC IgM-, IgG-, and IgA-plaque-forming cells could be demonstrated in spleen, bone marrow, lymph nodes, and Peyer's patches.     

Adjuvant Action of Capsular Polysaccharide of Klebsiella pneumoniae on Antibody Response

The sequence of histological changes in the regional lymph node and other lymphoid organs of mice injected with the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) or bacterial lipopolysaccharide (LPS) was followed. Injection of CPS-K, but not LPS, induced the following characteristic histological changes in the regional lymph node. In the early stage there was a marked decrease in the number of small lymphocytes, accompanied by the appearance of scattered fragmented nuclei and infiltration of polymorphonuclear neutrophilic leukocytes, and in the late stage there was marked proliferation of macrophage-like cells and pyroninophilic cells. Histological changes in the thymus and spleen and changes in cell populations in the bone marrow and peripheral blood after CPS-K injection were essentially the same as after LPS injection. Since CPS-K has a much stronger adjuvant action on antibody response than does LPS, it is suggested that the characteristic histological changes in the regional lymph node after injection of CPS-K are closely related to its extraordinarily strong adjuvant action.