T and B Lymphocyte Populations in Uterus Draining Lymph Nodes at Estrus and Diestrus in Wistar Rats

ABSTRACT: In this paper, data are presented on the characterization of uterous draining lymph nodes (UDLN) B and T lymphocytes of virgin rats at estrus (E) and diestrus (D). We established that the T/B lymphocyte relationship in the UDLN is less than one at estrus and more than one at diestrus. This is due to a decrease in the percentage and in the total number of mature T cell population in association with a decrease in the percentage of the OX8 subset in the UDLN at estrus. This situation may be related to an increase in estrogens known to produce thymic involution. These changes were not observed when we studied peripheral (popliteal) lymph nodes. The changes observed in the UDLN T cell population at estrus could be under hormonal control and we think that this condition may be important to prepare the immune system for an eventual pregnancy.     

An Unusual Case Involving the Spleen and Lymph Nodes

A case of peliosis hepatis in a 52-year old woman with unusual involvement of the spleen and lymph nodes is described. The initial sign was intraperitoneal hemorrhage from the splenic lesion, recessitating splenectomy. The hepatic lesion, which was first noticed during the operation, rapidly progressed and eventually resulted in hepatic rupture with fatal intraperitoneal hemorrhage. Serial sections treated by silver impregnation revealed degeneration and dissolution of the fine reticulin framework in the involved organs, suggesting the possible morphogenesis of the peliotic lesion. The patient had no history of any underlying disorders or of medication with steroids. Acta Pathol. Jpn. 32: 212∼215, 1989.     

Cytokine mRNA in the Joints and Draining Lymph Nodes of Rats with Adjuvant Arthritis and Effects of Cyclosporin A

TNF- and IL-1 promote leukocyte recruitment to arthritic joints and may contribute to cartilage degradation while regulatory cytokines such as IL-4 and IL-1RA may in part determine the course of arthritis. Here we report the pattern of TNF-, IL-1, IL-6, IFN-, IL-1RA, and IL-4 mRNA expression, detected by RT/PCR, in the talar joint and draining popliteal lymph node (PLN) of rats with adjuvant arthritis (AA). Levels of TNF- and IFN- mRNA were increased in the PLN before clinical signs of arthritis. This was followed by increases in IL-1 and IL-1RA mRNA at d9 and IL-6 mRNA at d12. PLN IL-1RA mRNA levels were positively correlated with those of IL-1 and TNF- throughout d5-d20. IL-4 mRNA levels were highest on days 7 and 20. In the synovium, a small increase in TNF-, IL-1, and IL-6 mRNA was detected on d5 then again on d12. Maximal synovial TNF- levels were reached on d20, while IL-1 peak expression was on d16 and IL-6 on d14. IL-4, IL-1RA, and IFN- mRNA was undetectable in the synovium. Cyclosporin treatment for 4 days, initiated at the height of arthritis, rapidly decreased clinical disease, and decreased migration of neutrophils and T lymphocytes into the joints. Yet no significant effect of CyA was observed on inflammatory cytokine expression, although the correlation between PLN IL-1RA and IL-1 or TNF- was lost in treated animals. Thus there is a variable pattern of cytokine gene expression in rat AA, the undetectable IL-4 and IFN- mRNA in synovium being analogous to human rheumatoid arthritis.     

Electrophoretic Mobility of Lymphocyte Populations in Juvenile Rheumatoid Arthritis

Peripheral blood lymphocytes from 16 patients with juvenile rheumatoid arthritis and 27 age-matched healthy controls have been studied, using several lymphocyte markers: electrophoretic mobility (EM), E-rosettes, immunofluorescence, and refringency. Eight patients (mean age, 6 years) were selected with a typical EM pattern--that is, a decrease in the mean EM of T cells and increase in B versus T-cell ratio. The other group of patients (mean age, 11 years) showed no significant difference when compared with their age-matched controls, with the exception of the positive refringence test. These findings suggest an impairment in the maturation of the immune system in childhood, which in turn may be responsible for the increased susceptibility to disease.     

Reactive Proliferative Lesions in Lymph Nodes from Rheumatoid Arthritis Patients

In 22 cases of rheumatoid arthritis (RA), including 4 cases of malignant RA (MRA), reactive proliferative lymph node lesions were studied clinicopathologically and immunohis-tochemically. This series included 5 males and 17 females. The period between disease onset and lymph node biopsy ranged from 3 months to 41 years. Generalized lymphadenopathy was noted in 13 cases and constitutional symptoms in 8. The histological findings characteristic of RA were 1) follicular hyperplasia with active germinal centers and 2) polyclonal plasma cell infiltration in the interfollicular area. Studies of intracytoplasmic immunoglobulin showed that γ-heavy chain-expressing plasma cells were a major component in the interfollicular area in 17 RA cases. However, in 4 MRA cases, a prominent increase of μ chain-expressing plasma cells was recognized in the same area. In the 3 cases for which fresh tissue sections were stained with monoclonal antibodies against lymphocytes, we found that the majority of T cells in the interfollicular area had helper/inducer markers. The identical locations of the T cell population and plasma cells indicated that both played a role in the proliferation and/or differentiation of B cells in lymph nodes in RA.     

Lymphocyte Populations and Cellular Immune Reactions in Juvenile Rheumatoid Arthritis

Ten patients with juvenile rheumatoid arthritis and age- and sex-matched healthy controls were investigated in pairs. The patients were found to have both normal proportions and normal absolute numbers of T lymphocytes, B lymphocytes, and the Fc-receptor-bearing lymphoid cells in peripheral blood. No abnormality of mitogen-induced lymphocyte transformation was observed. Lymphocyte-mediated cytotoxicity induced by phytohemagglutinin (PHA) or anti-target cell antibodies was also found to be normal. As in an earlier study, impaired delayed hyper-sensitivity by skin testing was observed in the patient group, thus indicating a dissociation between in vivo and in vitro parameters of lympboid cell function.     

Lymphocyte Populations and Cellular Immune Reactions in Juvenile Rheumatoid Arthritis

Ten patients with juvenile rheumatoid arthritis and age- and sex-matched healthy controls were investigated in pairs. The patients were found to have both normal proportions and normal absolute numbers of T lymphocytes, B lymphocytes, and the Fc-receptor-bearing lymphoid cells in peripheral blood. No abnormality of mitogen-induced lymphocyte transformation was observed. Lymphocyte-mediated cytotoxicity induced by phytohemagglutinin (PHA) or anti-target cell antibodies was also found to be normal. As in an earlier study, impaired delayed hyper-sensitivity by skin testing was observed in the patient group, thus indicating a dissociation between in vivo and in vitro parameters of lymphoid cell function.     

Nitric Oxide Modulates Lymphocyte Proliferation But Not Secretion of IL-2

The objective of this study was to determine the effects of nitric oxide (NO) on lymphocyte proliferation and cytokine release. Bronchoalveolar lavage (BAL) cells served as the source of NO and were obtained from rats treated with a single, intratracheal dose of bleomycin (3.6 mg/kg). At the time of sacrifice, the spleens were removed and the lymphocytes separated. Co-cultures containing BAL cells, lymphocytes and concanavalin-A were established and incubated at 37°C for 24 hours at which time proliferation, nitrite concentration and interleukin-2 (IL-2) production were measured. At ratios from 5:1 to 1:4 (BAL:lymphocyte) there was a significant reduction in lymphocyte proliferation. There was a significant, negative correlation between NO concentration and thymidine incorporation which was reversed when the NO synthase inhibitor N^G-monomethyl-L-arginine (NMA) was added to the co-cultures. Despite marked inhibition of spleen lymphocyte proliferation by NO₂ released by BAL cells, there was no corresponding reduction in IL-2 production. These data demonstrate that macrophages, activated in vivo, produce NO which regulates lymphocyte growth but not necessarily functions such as the secretion of the cytokine IL-2. Further, the ability of IL-2-dependent CTLL-2 cells to proliferate in the presence of excess IL-2 was also inhibited by BAL cells, confirming that NO inhibits lymphocyte growth.     

Morphogenesis of the Spleen During the Human Embryonic Period

We aimed to observe morphological changes in the spleen from the emergence of the primordium to the end of the embryonic period using histological serial sections of 228 samples. Between Carnegie stages (CSs) 14 and 17, the spleen was usually recognized as a bulge in the dorsal mesogastrium (DM), and after CS 20, the spleen became apparent. Intrasplenic folds were observed later. A high‐density area was first recognized in 6 of the 58 cases at CS 16 and in all cases examined after CS 18. The spleen was recognized neither as a bulge nor as a high‐density area at CS 13. The mesothelium was pseudostratified until CS 16 and was replaced with high columnar cells and then with low columnar cells. The basement membrane was obvious after CS 17. The mesenchymal cells differentiated from cells in the DM, and sinus formation started at CS 20. Hematopoietic cells were detected after CS 18. The vessels were observed at CS 14 in the DM. Hilus formation was observed after CS 20. The parallel entries of the arteries and veins were observed at CS 23. The rate of increase in spleen length in relation to that of stomach length along the cranial‐caudal direction was 0.51 ± 0.11, which remained constant during CSs 19 and 23, indicating that their growths were similar. These data may help to better understand the development of normal human embryos and to detect abnormal embryos in the early stages of development. Anat Rec, 298:820–826, 2015. © 2014 Wiley Periodicals, Inc.     

Size, CD4 and CDS Marker Profiles and Functions of Lymphocyte Subpopulations in Mucosal-Associated Lymph Nodes

We analyzed phenotypic and functional characteristics of T cell populations in mucosal-associated supramammary and mesenteric lymph nodes in goats. Here we demonstrate, by flow cytometry, quantitative differences in CD4 and CD8 T cell subsets among large and small mucosal-associated lymphocyte populations and their differential regulatory activities on resident lymph node B cells stimulated with Staphylococcus aureus Cowan I or pokeweed mitogen. The CD4/CD8 T cell ratio was Tower in mesenteric lymph nodes (1.46) when compared to that of supramammary lymph nodes (2.18), Analysis of large and small lymphocyte subpopulations from lymph nodes showed nearly 62% of the lymphocytes from mesenteric lymph nodes being of large cell phenotype with CD4/CD8 ratios of 1.34. In contrast, large cell subpopulations in supramammary lymph nodes showed a significantly lower number (50%) with a higher CD4/CD8 ratio of 2.05. Functionally, mesenteric lymph node T cells, isolated by nylon wool, showed heightened suppressive activity in mitogen-driven B cell proliferation responses, whereas T cells from supramammary lymph nodes were stimulatory. These findings clearly demonstrate distinctive functional properties between resident T cell populations of supramammary and mesenteric lymph nodes, suggesting that different proportions of T cell subsets in these; nodes are activated and thus regulate regional immune responses via different pathways.     

Size,CD4 and CDS Marker Profiles and Functions of Lymphocyte Subpopulations in Mucosal-Associated Lymph Nodes

We analyzed phenotypic and functional characteristics of T cell populations in mucosal-associated supramammary and mesenteric lymph nodes in goats. Here we demonstrate, by flow cytometry, quantitative differences in CD4 and CD8 T cell subsets among large and small mucosal-associated lymphocyte populations and their differential regulatory activities on resident lymph node B cells stimulated with Staphylococcus aureus Cowan I or pokeweed mitogen. The CD4/CD8 T cell ratio was Tower in mesenteric lymph nodes (1.46) when compared to that of supramammary lymph nodes (2.18), Analysis of large and small lymphocyte subpopulations from lymph nodes showed nearly 62% of the lymphocytes from mesenteric lymph nodes being of large cell phenotype with CD4/CD8 ratios of 1.34. In contrast, large cell subpopulations in supramammary lymph nodes showed a significantly lower number (50%) with a higher CD4/CD8 ratio of 2.05. Functionally, mesenteric lymph node T cells, isolated by nylon wool, showed heightened suppressive activity in mitogen-driven B cell proliferation responses, whereas T cells from supramammary lymph nodes were stimulatory. These findings clearly demonstrate distinctive functional properties between resident T cell populations of supramammary and mesenteric lymph nodes, suggesting that different proportions of T cell subsets in these; nodes are activated and thus regulate regional immune responses via different pathways.     

Immune Response in Deep Cervical Lymph Nodes and Spleen in the Mouse after Antigen Deposition in Different Intracerebral Sites

Brain interstitial and cerebrospinal fluid drainage into the lymphatics was studied by injections of 5 microliters of packed sheep red blood cells (SRBC) injected into the caudate nucleus, the occipital lobe, and the lateral ventricle of the brain in mice. The number of plaque-forming cells (PFC) was determined in the deep cervical lymph nodes, the axillary lymph nodes, and the spleen, and the number of PFC was compared with the response in the same tissues after intravenous immunization with 0.1 ml 10% SRBC. The weight of the deep cervical lymph nodes increased 3.0 times on day 3 after injection in the brain parenchyma compared with the weight of these nodes after intravenous immunization. The antigen-specific response peaked on day 5, 392 +/- 37 PFC/10(6) for IgG in the deep cervical lymph nodes after antigen deposition in the caudate nucleus, whereas only a minor peak in the antigen-specific response was obtained after intraventricular antigen deposition, 127 +/- 79 PFC x 10(6) for IgG on day 6. There were no increased PFC in any of the lymph nodes after intravenous immunization. The experiments show an antigen-specific response in the deep cervical lymph nodes after intracerebral antigen deposition, whereas antigens deposited in the lateral ventricles drain preferentially to the blood, with a high response in the spleen.     

Opposite Effects of BCG on Spleen and Lymph Node Cells: Lymphocyte Proliferation and Immunoglobulin Synthesis

C57BL/6 mice were immunized intravenously (i.v.), intraperitoneally (i.p.), or subcutaneously with one dose of Bacillus Calmette-Guérin (BCG). At various time intervals after injection, the lymphocyte response, as measured by thymidine incorporation into DNA, and the number of immunoglobulin-secreting cells were determined in vitro before and after mitogenic stimulation with phytohemagglutinin, concanavalin A, or lipopolysaccharide. In unstimulated cultures, the spontaneous thymidine incorporation and immunoglobulin synthesis of spleen cells were increased to some extent in mice infected i.p. or i.v. with BCG, as compared with noninfected mice. In contrast, after mitogenic stimulation, a marked depression of the proliferative response of spleen cells to both T- and B-cell mitogens and a marked inhibition of LPS-induced immunoglobulin secretion were observed in mice infected i.v. and to a lesser extent in those infected i.p. The depression of lymphoblastogenesis in spleens was fully established 15 days after infection and persisted for a long period of time. When unfractionated or plastic-adherent spleen cells from BCG-infected mice were cultured with normal spleen cells, a strong depression of their reactivity to phytohemagglutinin, concanavalin A, and lipopolysaccharide was observed. After the removal of cells adherent to plastic, the response was partially restored in the nonadherent population from mice infected i.p., but not in that from mice infected i.v. After mitogenic stimulation, lymph node cells of mice inoculated subcutaneously showed a response to mitogen higher than that of normal cells. These results thus demonstrate that, depending on the route of administration, BCG exerts very different effects.     

Ultrastructural Analysis of Antibody Synthesis in Cells from Lymph and Lymph Nodes

A variety of cells containing antibody were found in lymph from sheep responding to secondary challenges with horseradish peroxidase. Antibody was present in blast cells in lymph within the perinuclear space, the endoplasmic reticulum, the Golgi apparatus and on polyribosomes. Some lymphocytes in lymph also contained antibody but in these cells it was located principally in the perinuclear space. No cells were found containing antibody distributed throughout their cytoplasm nor were any lymphocytes found with a Golgi apparatus positive for antibody. After the immune response in the lymph had died away, a population of cells containing antibody was still present in the regional lymph node. These were all plasma cells in which the antibody was present for the most part in a highly organized endoplasmic reticulum. Cells of this type were never found in the lymph. The cells containing the smallest amounts of antibody had a few discrete focal points in the perinuclear space and a few positive groups of ribosomes in their cytoplasm. The endoplasmic reticulum in some cells was filled completely with antibody, while in others positive segments were found adjacent to negative ones. The antibody in the cytoplasmic endoplasmic reticulum was in continuity with the antibody in the perinuclear space. The Golgi apparatus contained antibody in only a small proportion of the cells but when it was positive it was strongly so, suggesting that antibody was concentrated in this organelle. In some cells a positive reaction to horseradish peroxidase antibody appeared in the nucleus over the nucleoli. The significance of this finding is not known.