Lymphocyte subpopulations in sensitization and specific immunotherapy in an experiment. Lymphocyte subpopulations in sensitization to staphylococci, Artemisia pollen and their combinations

The distribution of T- and B-lymphocytes in the body of guinea pigs was studied in different groups of the animals. As shown in this study, in delayed hypersensitivity to staphylococci the number of PE- and E-rosette-forming cells increased in the blood, the spleen, and the lymph nodes and decreased in the thymus; the number of EA- and EAC-rosette-forming cells decreased in the bone marrow and the spleen, the number of T gamma-suppressors decreased in the bone marrow and the distant lymph node. Immediate hypersensitivity to tarragon pollen induced the general increase of the content of T- and B-lymphocytes; the number of T gamma-cells decreased in the thymus, the bone marrow, and the lymph nodes and increased in the spleen. The characteristic features of combined microbial-pollen sensitization were the high content of B-cells in all lymphoid organs (except the thymus), a low level of T-lymphocytes in the blood and the peripheral lymphoid organs, the decreased number of T gamma-cells in most of the immunogenetic organs.     

Comparative study of the T- and B-cell immunity systems in human and animal embryogenesis

Data are presented on the spleen and thymus structure, time of appearance of the lymphocytes and their heterogeneity in the liver, thymus, spleen, lymph nodes and blood of human foetuses with hemochorial placenta (3 to 34 weeks) and of the minipigs foetuses with epitheliochorial placenta (32 to 95 days). In both foetuses the first T- and B-lymphocytes are found in liver, T-lymphocytes are then found in thymus and later in spleen and lymph nodes whereas B-lymphocytes are found, after liver, in spleen. Kinetics of T- and B-lymphocytes during embryogenesis is described. Reaction of the minipig lymphocytes to mitogens was demonstrated.     

Effect of the antineoplastic antibiotic bruneomycin on lymphocyte population ratio in mice

Data on the quantitative ratio of the populations of T- and B-cells in the lymphoid organs of mice at various periods after oral administration of bruneomycin are presented. It was shown that in contrast to the total number of T- and B-cells in the spleen and mesenteric lymph nodes amounting to the minimal values at the early periods (days 1-3) after the antibiotic injection, their relative content remained rather high. Complete recovery of the number of T- and B-lymphocytes in the above organs was observed only by the end of a month period. Study on the kinetics of the immune response to sheep red blood cells showed that reactivity of the antibody producers in the experiments with bruneomycin increased 1.5-2 fold as compared to the control.     

Lymphocyte subpopulations in sensitization and specific immunotherapy in an experiment. Lymphocyte subpopulations in the specific immunotherapy of microbial allergy and subsequent heterologous pollen sensitization

The influence exerted by the specific immunotherapy (SIT) of delayed hypersensitivity (DH) to staphylococci and subsequent sensitization with tarragon pollen on the level of immunocompetent cells in the blood and lymphoid organs of guinea pigs was studied. On the whole, SIT normalized the characteristics of T- and B-lymphocytes, altered as the result of experimentally induced DH: the content of T-cells in the peripheral blood and the lymph nodes increased, while the number of B-cells in the blood and T gamma-suppressors increased. The subsequent heterologous sensitization with pollen abolished the effect of SIT, inducing the general decrease of the level of T gamma-lymphocytes and enhancing the number of T-lymphocytes in the lymph nodes.     

Change in the strength of DNA-protein binding in T- and B-lymphocytes of the spleen of C3HA mice during chemical hepatocarcinogenesis

The tightness of DNA-protein binding in the nuclei of mouse spleen T- and B-lymphocytes was assessed, using nucleoprotein celite chromatography, and changes in the number of T- and B-suppressors in the course of o-AAT-induced chemical hepatocarcinogenesis were studied. Attenuation of DNA-protein bonds in T-lymphocytes at the early stages (up to 3 months) was observed, and by the time of hepatoma formation (8 months) about 50% of T-lymphocyte DNA was loosely bound to proteins, which is a typical feature of quiescent cells. In B-lymphocytes attenuation of DNA-protein interaction was only observed by the 8th month of carcinogenesis. By the time of hepatoma formation the number of T-suppressors in mouse spleen increased 2.8-fold, while the number of B-suppressors in lymph nodes remained unchanged.     

Participation of mouse T- and B-lymphocytes in the graft versus host reaction]

Cells of the spleen or lymph nodes of CBA mice were transplanted to sublethally irradiated (CBAXC57BL/6)F1 mice; this caused development of the graft-versus-host reaction (GVHR). Lymphocytes lost the capacity to realize this reaction after in vitro treatment with specific sera against mouse T- and B-lymphocytes. Apparently, development of the GVHR in mice was connected with the cooperative interaction of T- and B-lymphocytes.     

Migration patterns of dendritic cells in the rat: comparison of the effects of gamma and UV-B irradiation on the migration of dendritic cells and Lymphocytes

To further define the underlying mechanisms of immune suppression induced by UV-B irradiation, we have examined the kinetics of homing patterns of in vitro UV-B-irradiated and gamma-irradiated-thoracic duct lymphocytes (TDL) compared to dendritic cells (DC). Our findings show that 111In-oxine-labeled TDL specifically home to the spleen, liver, lymph nodes, and bone marrow with subsequent recirculation of a large number of cells from the spleen to lymph nodes. In contrast, DC preferentially migrate to the spleen and liver with a relatively insignificant distribution to lymph nodes and an absence of subsequent recirculation. Splenectomy prior to cell injection significantly diverts the spleen-seeking DC to the liver but not to the lymph nodes, while the homing of TDL to lymph nodes is significantly increased. In vitro exposure of 111In-oxine labeled TDL to gamma irradiation does not significantly impair immediate homing to lymphoid tissues but inhibits cell recirculation between 3 and 24 hr. In contrast, gamma irradiation does not affect the tissue distribution of labeled DC, suggesting that DC are more radioresistant to gamma irradiation than TDL. Unlike the findings in animals injected with gamma-irradiated cells, UV-B irradiation virtually abolished the homing of TDL to lymph nodes and significantly reduced the homing of the spleen-seeking DC to the splenic compartment while a large number of cells were sequestered in the liver. The results of in vitro cell binding assay show that TDL, unlike DC, have the capacity to bind to high endothelial venules (HEV) within lymph node frozen sections while gamma and UV-B irradiation significantly inhibit the binding of TDL to lymph node HEV. These findings suggest that: (i) DC, unlike TDL, are unable to recirculate from blood to lymph nodes through HEV; (ii) although gamma irradiation impairs TDL recirculation, it does not affect DC tissue distribution; and (iii) UV-B irradiation impairs both TDL and DC migration patterns. We conclude that the lack of capacity of irradiated TDL to home to lymph nodes is due to damage to cell surface homing receptors and that the failure of DC to home to the lymph node microenvironment is related to the absence of HEV homing receptors on their cell surface.     

The entry of T and B lymphocytes into rat popliteal lymph nodes undergoing a graft-versus-host reaction

The entry of radiolabeled blood-borne T and B lymphocytes into resting popliteal lymph nodes and popliteal lymph nodes stimulated with semiallogeneic lymphocytes was investigated in rats. Thoracic duct lymphocytes separated into T- and B-lymphocyte populations on nylon-wool columns were radiolabeled with 51chromium and equal numbers of T or B lymphocytes were injected intravenously. While the ratio of T and B lymphocytes in the blood is approximately 3:1 it was found that the ratio of T to B lymphocytes migrating into lymph nodes was approximately 9 T to 1 B lymphocyte in both resting and antigenically stimulated lymph nodes. Since the ratio of T to B lymphocytes in thoracic duct lymph is similar to that of blood, there is a disparity between the number of T cells entering and leaving lymph nodes. These results suggest that some T lymphocytes may return to the blood directly and/or there is increased T lymphocyte death in lymph nodes.     

Dynamics of immunoglobulins and immunocompetent lymphocytes in guinea pigs with experimental ascariasis

The paper presents data on variations in the level of serous immunoglobulins (Ig M and Ig G classes), immunocompetent T- and B-lymphocytes in lymphoid organs (spleen, mesenteric, portal and mediastinal lymph nodes) and specific antiparasitic antibodies in guinea pigs during the dynamics of experimental ascariasis.     

Nonrandom migration of CD4+, CD8+, and gamma delta+T19+ lymphocyte subsets following in vivo stimulation with antigen

We report here the results of experiments in which the migration of three T cell subsets (CD4+, CD8+, and gamma delta+T19+ cells) through antigen-stimulated lymph nodes and subcutaneous granulomas has been compared with that through normal skin and resting lymph nodes. The percentage of gamma delta+T19+ lymphocytes was halved and the percentage of CD8+ lymphocytes was doubled in lymph draining stimulated compared with control tissues, and all lymphocyte subsets except gamma delta+T19+ lymphocytes had higher hourly outputs in lymph draining antigen-stimulated compared with control tissues. Antigen also resulted in a higher percentage of CD8+ lymphoblasts and a lower percentage of gamma delta+T19+ lymphoblasts in efferent lymph draining antigen-stimulated lymph nodes. The data indicate that lymphocyte subsets leave the blood with differing efficiencies in different vascular beds and raise the possibility that antigen can influence the rate at which tissues extract individual T cell subsets from the blood.     

Y+- and L-system amino acid transport in normal and chronic lymphocytic leukemia lymphocytes: photoinhibition by fluoronitrophenylazide

Three major pathways mediate amino acid transport into mammalian cells: the A-system and the ASC-system, which require a sodium gradient across the plasma membrane, and the L-system, which has no requirement for a sodium gradient. We have found that the lymphocytes from patients with B-cell chronic lymphocytic leukemia (CLL) have a marked reduction in the L-system of amino acid transport when compared to normal human B-lymphocytes from blood or tonsils. Transport by the A- and ASC-systems was not decreased in CLL B-lymphocytes. Because of the specific defect of the sodium-independent L-system amino acid transport in CLL cells, we have examined the activity of another sodium-independent transport system, the Y+-system, in human lymphocytes. The Y+-system favors the transport of dibasic, cationic amino acids such as lysine, ornithine, and arginine, which carry a positively charged group on their side chains. Our studies indicate that there is a large nonsaturable component of amino acid transport by the Y+-system in human lymphocytes. Using a multicomponent mathematical analysis, we have determined that the saturable component of Y+-transport is similar in T- (thymus-derived) and B- (bone-marrow-derived) lymphocytes and is unimpaired in CLL B-lymphocytes. Further, fluoronitrophenylazide, which was thought to be a specific inhibitor of the Y+-system when photoactivated, also inhibits A-, and L-system transport in CLL, T-, and B-lymphocytes.     

Characteristics of individual lymphocyte populations in the blood of virtually healthy persons]

The number of individual lymphocyte populations in the blood of apparently healthy persons was determined by spontaneous rosette-formation with SRBC (T cells) and mouse red blood cells (B cells). In the majority of persons examined the percentage of T-lymphocytes constituted 47.3 +/- 1,6, of B--16.8 +/- 1, and of "zero" cells--33.5 +/- 3.4. There was less T- and B-lymphocytes in the blood of elderly persons (over 50 years of age) than in the young ones. It was also shown that the T-lymphocyte population forming "active" rosettes could be assessed by the number of SRBC sorbed on their surface.     

Cyclic adenosine 3',5'-monophosphate response to dopamine in mouse lymphoid cells and cell lines

Cyclic adenosine 3',5'-monophosphate responses to dopamine and isoproterenol were studied in mouse and rat spleen, thymus, lymph nodes and Peyer's patches lymphocytes and in 7 mouse cell lines of T- and B-lymphoid derivation. The responses of normal cells to dopamine were moderate, of the same extent, but selective to spleen and thymus in mouse, and to spleen and lymph nodes in rat. The YAC-1 T lymphoma cell line was sensitive to dopamine with a higher magnitude than normal lymphoid cells. Dopamine was less potent than isoproterenol in all cells, and whereas dopamine-sensitive and isoproterenol-sensitive cells, or dopamine-insensitive and isoproterenol-insensitive cells were found, no cell type was dopamine-sensitive and isoproterenol-insensitive. Altogether, these results suggest that only a small subset of lymphocytes is susceptible to the cAMP-elevating action of dopamine.     

Characteristics of the lymphoid cells of rabbits]

By means of scanning electron microscopy it has been stated that the thymus gland, spleen, paratracheal lymph node, Peyer's patches and appendix in an intact rabbit of "Shinshilla" strain have T- and B-lymphocytes in different proportion. T-lymphocytes are seen as spherical cells with rough surface, with a few microprocesses, have 3.0 +/- 0.1 mcm--4.7 +/- 0.2 mcm in diameter depending on their location. B-lymphocytes are seen as spherical cells with a considerable amount of microvilli on their surface, have 2.9 +/- 0.1 mcm--4.4 +/- 0.1 mcm in diameter depending also on their location. In the thymus gland, cells with rough surface, a few microprocesses and crater-like hollows prevail. In the spleen and lymph node, most of the cells are of similar diameter and have microvilli on the surface. Lymphoid population in the Peyer's patch and appendix is represented by large cells with microprocesses.     

Mechanisms of action of synthetic polyelectrolytes and polyampholytes on the immune response]

We investigated the effect of polyacrilic acid (PAA) on the immune response in mice of various strains on sheep red blood cells and also the influence of poly-2-methyl-5-vinyl-pyridine (PMVY), PAA and their statistical copolymers on antibody-forming cells (AFC) production in cultures of T- and B-lymphocytes in vivo. PAA was seen to increase accumulation of AFC in the spleen of mice depending on their genotypes. PMVP and PAA were found to intensify the cooperating interaction of T- and B-lymphocytes, whereas their copolymers exert quite an opposite effect. The injection of copolymers to the recipients of cooperating T- and B-lymphocytes practically results in the complete elimination of the cooperation effect between T- and B-lymphocytes in the immune response to sheep erythrocytes without cytostatic action of cell proliferation.     

CD8+ alpha/beta or gamma/delta T cell receptor-bearing T cells from athymic nude mice are cytolytically active in vivo.

Phenotypic analysis of lymphocytes that mature extrathymically in congenitally athymic nude mice has revealed a large population of CD3+ CD8+ T cells that express gamma/delta-TCR. In euthymic mice, significant numbers of cells with this phenotype are found only in the intestinal epithelium. Intestinal intraepithelial lymphocytes have been shown to be cytolytically active in vivo, as measured by the redirected lysis assay. In this communication, freshly harvested T cell subsets obtained from pooled nude mouse spleen and lymph nodes and separated by flow cytometric cell sorting were assayed for their ability to lyse FcR+ P815 targets in the presence of mAb to the epsilon-chain of the CD3 complex. CD8+, but not CD4+ or CD4- CD8-, T cells in nude mice were cytolytically active. CD8+ alpha/beta- and gamma/delta-TCR-bearing T cells from the spleen and lymph nodes of nude mice demonstrated similar cytolytic activity. No cytolytic activity of purified cell subsets was apparent in the absence of anti-CD3 mAb, even when NK-susceptible target cells were used. These data indicate that, in contrast to euthymic mice, a large proportion of CD8+ cells from the spleen and lymph nodes of nude mice are cytolytically active in vivo. In addition, these results suggest that the intestinal epithelium is not the only anatomical location where constitutively cytolytic CD8+ alpha/beta- or gamma/delta TCR-bearing T cells may be found.     

Vimentin function in lymphocyte adhesion and transcellular migration

Although the adhesive interactions of leukocytes with endothelial cells are well understood, little is known about the detailed mechanisms underlying the actual migration of leukocytes across the endothelium (diapedesis). Leukocytes have been shown to use both paracellular and transcellular routes for transendothelial migration. Here we show that peripheral blood mononuclear cells (PBMCs; T- and B-lymphocytes) preferentially use the transcellular route. The intermediate filaments of both endothelial cells and lymphocytes formed a highly dynamic anchoring structure at the site of contact between these two cell types. The initiation of this process was markedly reduced in vimentin-deficient (vim(-/-)) PBMCs and endothelial cells. When compared with wild-type PBMCs, vim(-/-) PBMCs showed a markedly reduced capacity to home to mesenteric lymph nodes and spleen. Furthermore, endothelial integrity was compromised in vim(-/-) mice, demonstrating that intermediate filaments also regulate the barrier that governs leukocyte extravasation. Absence of vimentin resulted in highly aberrant expression and distribution of surface molecules critical for homing (ICAM-1 and VCAM-1 on endothelial cells and integrin-beta1 on PBMCs). These data show that intermediate filaments are active in lymphocyte adhesion and transmigration.     

PHA activation of encephalitogenic T cells: in vitro line selection overcomes splenic suppression

In this study we compared myelin basic protein (MBP) and phytohemagglutinin (PHA) for their ability to induce proliferation and experimental autoimmune encephalomyelitis (EAE) transfer activity in mixed cell cultures obtained from spleen and lymph nodes versus highly selected MBP-specific T cell lines and clones. Established MBP-specific cells derived initially from immune lymph nodes attained both proliferative and EAE-transfer activities after in vitro activation with either MBP or PHA. In contrast, PHA was unable to induce immune spleen cells to transfer EAE, in spite of its potent mitogenic activity. On the basis of these results, we evaluated the in vitro proliferation and differentiation responses of MBP-specific T cells during the line selection process using cells derived from both immune lymph node and immune spleen. During the initial selection process with MBP, proliferation of MBP-specific T cell precursors from immunized spleen populations was reduced relative to lymph node cells. After antigen-dependent selection the encephalitogenic cells from either organ exhibited identical in vitro response characteristics. Freshly isolated immune spleen cells were potent suppressors of MBP-specific T cell proliferation suggesting that the in vitro differences between the two organs was due to splenic suppression of the encephalitogenic cells.     

Are γδ T cells important for the elimination of virus-infected cells?

Rhesus monkeys (Macaca mulatta) gamma delta T cells were identified using a monoclonal antibody. The relative representation of gamma delta T lymphocytes in the peripheral blood, lymph nodes, and spleen resembles that of Homo sapiens. The analysis of function and specificity revealed further significant similarities between the simian and human gamma delta T-cell systems. Since both human and monkey gamma delta T lymphocytes can effectively lyse cells infected with immunodeficiency viruses, it is possible that the primate gamma delta T-cell systems contribute to antiviral immunosurveillance.