Association between T lymphocyte sub-sets apoptosis and peripheral blood mononuclear cells oxidative stress in systemic lupus erythematosus

Abstract

Increased oxidative stress and lymphocyte apoptosis are a hallmark of the autoimmune disease systemic lupus erythematosus (SLE). However, the association between oxidative stress and T lymphocytes apoptosis has still to be elucidated in SLE. In order to appraise the interaction between oxidative stress and T lymphocyte apoptosis with the severity of disease, oxidative stress profile and T lymphocytes apoptosis were studied. Increased levels of ROS, MDA and CD4⁺ lymphocyte apoptosis were positively associated with disease activity while decreased levels of GSH and percentage expression of CD4⁺ lymphocyte were negatively associated with disease activity. The decrease in intracellular levels of GSH was negatively associated with T lymphocyte, CD4⁺ lymphocyte, CD8⁺ lymphocyte apoptosis and intracellular caspase-3 expression. The present study suggests that increased T lymphocyte sub-sets apoptosis may be mediated by decreased intracellular glutathione concentration and severity of disease might be enhanced together by over-production of ROS in SLE.     

Thymus and Bursa Dependence of Lymphocyte Mitogenic Factor in the Chicken

AMONG the soluble products generated during activation of rodent or human lymphocytes¹, lymphocyte-stimulating (‘mitogenic’) factors are detected by their ability to accelerate DNA metabolism of freshly cultured allogeneic or syngeneic lymphocytes^2–8. If such lymphocyte activation products function as mediators of cellular immunity^1,9–11, their activities will be generated independently of the antibody-forming system. In the chicken, antigen-stimulation of sensitized lymphocytes in vitro involves both thymus (T-) and bursa-(B-)-dependent cells^12–14. If antigen-induced lymphocyte transformations are generally mediated by lymphocyte mitogenic factors, both T-cells and B-cells should produce lymphocyte mitogenic factors when stimulated by specific antigen. We have therefore determined whether antigen-stimulated chicken lymphocytes generate a lymphocyte mitogenic factor and whether this response is affected by neonatal thymectomy or bursectomy.     

Flow cytometric characterization of the lymphocyte composition in a variety of mucosal tissues in healthy rhesus macaques

Background Rhesus monkeys play a central role in model studies on human infectious diseases, and often mucosal organs are affected by these pathogens, e.g. HIV. However, a comparative investigation into lymphocyte composition from different mucosal tissues is still missing. Methods Lymphocyte composition of duodenum, jejunum, ileum, colon, vagina, cervix, uterus and bronchoalveolar lavage from healthy rhesus monkeys was characterized in detail by flow cytometry. Moreover, we compared the lymphocyte proportions from intestinal biopsies with resections. Results All mucosal tissues exhibited higher values of CD8⁺, CD4⁺ CCR5⁺ and CD45RA^? memory T cells than blood, but similar levels of total T cells. Especially within the four gut sites, the lymphocyte composition varied significantly. The relative proportions of lymphocyte subsets from duodenal and colonic biopsies compared to resections differed. Conclusion The lymphocyte composition highly varies between different mucosal sites, and data obtained from biopsy and necropsy samples were mostly not comparable.     

Phorbol myristate acetate stimulation of lymphocyte guanylate cyclase

Human lymphocyte guanylate cyclase activities are increased in a dose-dependent fashion by incubation of intact cells with phorbol myristate acetate, a tumor promoter and lymphocyte mitogen. Increased activity is detectable after 1 minute, and peak membrane-bound and soluble forms of guanylate cyclase occur after 10- and 30-minute exposure to phorbol myristate acetate, respectively. The soluble form is stimulated much more than the membrane form. Enzyme activities measured in the presence of either Ca²⁺, Mg²⁺, or Mn²⁺ are elevated to similar degrees. Comparisons of phorbol and a series of its diesters revealed a good correlation between the capacities for guanylate cyclase stimulation, lymphocyte mitogenesis, and tumor promotion.     

Lymphocyte–Antigen Interaction in Electrophoretic Mobility Test for Cellular Sensitization

A SENSITIVE electrophoretic method for measuring lymphocyte sensitivity has been developed and applied to the study of human disease^2,3 (including cancer⁴) and to the mode of action of antilymphocytic serum⁵. In principle, the method depends on the observation that a sensitized lymphocyte interacts with its specific antigen (but not with others) to liberate into the ambient medium some material which has the property of causing electrophoretic slowing of normal macrophages. The latter are used merely as an indicator system of lymphocyte–antigen interaction.     

Influence of perioperative whole blood transfusions on lymphocyte subpopulations in patients with stage II breast cancer

Preliminary reports have suggested an adverse relationship between blood transfusion and survival after surgery in patients with solid tumour. One might postulate that from these studies, perioperative blood transfusions alter host immune defences. We therefore examined the influence of homologous whole blood transfusion on circulating lymphocyte subpopulations in transfused patients compared with non-transfused patients. Fifty-one women with Stage II breast cancer who underwent surgical procedures were studied. Patients were classified into two groups on the basis of whether or not they had received blood transfusion. The lymphocyte subpopulations were analyzed by flow cytometry before cancer surgery and three weeks after the operation. CD3⁺, CD4⁺, CD8⁺, and CD20⁺ cells as the lymphocyte subsets were quantitated using appropriate monoclonal antibodies. No significant differences between pre- and postoperative lymphocyte subset levels were seen in non-transfused patients. However, there was a statistically significant increase in the CD8⁺ cell count; decreasing CD4⁺ cell count and decreased CD3⁺ cells levels were observed in the transfused group (P<0.05). Although these early results of the study suggest that the blood transfusions could be associated with alterations in lymphocyte populations, additional studies are needed to elucidate the possible mechanism of the transfusion-induced immunological modulations.     

Selective expression of murine lymphocyte alloantigens controlled by the X-chromosome

Alloantisera specific to X-chromosome linked lymphocyte membrane antigens (Ly-X) were prepared by immunizing F1 male mice with identical F1 female lymphocytes. Independent B cell specific (anti Lyb-X) and T cell specific (anti Lyt-X) antibodies were detected. The Lyt-X antigen was expressed on Lyt-2⁺, 3⁺, and on Tla^–, Lyt-1⁺, 2⁺, 3⁺ T cell subpopulations. The problem of X-chromosome inactivation and the relationship ofH-2-linkedIr genes and Ia antigens, with X-linkedIr genes and lymphocyte alloantigens are discussed.     

Lymphocyte subset analysis for the assessment of treatment-related complications after autologous stem cell transplantation in multiple myeloma

Background aimsThe aim of this study was to investigate the correlation between infused lymphocyte populations and lymphocyte subsets at engraftment, and the early clinical implications of lymphocyte subset recovery after autologous stem cell transplantation (ASCT) in multiple myeloma (MM).MethodsWe examined the lymphocyte populations of infused autografts and the lymphocyte subsets of peripheral blood at engraftment from 50 patients using flow cytometry. Each subset was grouped as low (below median) and high (above median) to examine the correlation with mucositis of grade 3 or more and the occurrence of infections and cytomegalovirus (CMV) reactivation.ResultsUsing Spearman correlation coefficients, we found that cell doses of infused CD8⁺ (P = 0.042) and CD19⁺ cells (P = 0.044) were significantly associated with the absolute lymphocyte count (ALC) at engraftment. The dose of infused CD34⁺ cells was not associated with the change of lymphocyte subsets except for an inverse correlation with CD4⁺ cells (P = 0.006). After adjusting for potential variables in univariate analysis, multivariate analyzes revealed that the lower ratio of infused CD4⁺ to CD8⁺ cells (P = 0.030) was an independent factor for severe mucositis. Of lymphocyte subsets at engraftment, a higher frequency of CD3⁺ (P = 0.024) and a lower frequency of CD56⁺ (P = 0.020) were independent predictors for infections after engraftment. A higher frequency of CD8⁺ cells (P = 0.041) and a lower ratio of CD4⁺ to CD8⁺ (P = 0.021) were independent predictors for CMV reactivation.ConclusionsOur data suggest that lymphocyte subset analysis of infused autograft and peripheral blood at engraftment may provide new predictors for early complications after ASCT in patient with MM.     

黄芪多糖联合布洛芬对创伤感染患者巨噬细胞分泌功能、T淋巴细胞亚群水平的影响及其影响因素分析

目的:探讨黄芪多糖联合布洛芬对创伤感染患者巨噬细胞分泌功能、T淋巴细胞亚群水平的影响及影响因素。方法:2014年3月-2018年3月我院收治创伤感染患者85例,其中重度感染39例,将所有患者按感染轻重进行平衡随机分为对照组43例和研究组42例,对照组应用布洛芬治疗,研究组应用黄芪多糖联合布洛芬治疗。比较两组患者巨噬细胞分泌功能、T淋巴细胞亚群水平,分析患者发生重度感染的影响因素及巨噬细胞分泌功能、T淋巴细胞亚群水平与创伤感染的关系。结果:治疗后研究组患者血清中肿瘤坏死因子(TNF)、白介素细胞-6(IL-6)和白介素细胞-1(IL-1)的水平低于对照组(P<0.05)。治疗后研究组外周血中CD3⁺、CD4⁺、CD8⁺和CD4⁺/CD8⁺水平高于对照组(P<0.05)。巨噬细胞分泌功能较高、T淋巴细胞亚群水平较低的患者发生重度感染的概率较高(P<0.05);巨噬细胞分泌功能与创伤感染呈正向关联关系(P<0.05,OR>1);T淋巴细胞亚群水平与创伤感染呈负向关联关系(P<0.05,OR<1);治疗方法以研究组方法为优(P<0.05,OR>1)。结论:黄芪多糖联合布洛芬治疗创伤感染患者效果更显著,巨噬细胞分泌功能、T淋巴细胞亚群水平均是创伤感染的影响因素。     

Lymphocyte suppression associated with alpha globulin. Changes in cellular immunity

Sensitization of guinea pigs to 2,4-dinitrofluorobenzene is accompanied by increases in alpha globulins determined electrophoretically. During sensitization, lymphocyte responses were measured in vitro by mitogen induced ³H-thymidine uptake in whole blood cultures and in vivo by dermal skin reactivity. Following 5 days of dinitrofluorobenzene sensitization alpha globulins were elevated and lymphocyte transformation to phytohemagglutinin, pokeweed mitogen, and concanavalin A was significantly suppressed. When the alpha globulins returned to normal levels following sensitization, lymphocyte responses returned to pretreatment values. Antigen induced lymphocyte responsiveness was also suppressed concomitant with elevations in alpha globulins. Tuberculin sensitive guinea pigs responded poorly to PPD in vitro and in vivo during DNFB sensitization. It is suggested that increases in alpha globulins detected during the development of cellular immunity are associated with immunosuppression.     

Histocompatibility Gene Organization and Mixed Lymphocyte Reaction

TRANSFORMATION of allogenic lymphocytes in mixed cultures depends chiefly on an incompatibility between the lymphocyte donors at the major histocompatibility locus in man (HL-A), mouse (H-2) and rat (H-l)¹. Although the mouse H-2 locus can be divided into several regions each of which controls one or more antigenic specificities² and two or more subloci control HL-A antigens in man³, it is not known whether all parts of the major histocompatibility locus are equally important in eliciting transformation in mixed lymphocyte cultures. We now show that capacity to elicit lymphocyte transformation is different for different parts of the mouse H-2 locus.     

FOXP3+ Lymphocyte Density in Pancreatic Cancer Correlates with Lymph Node Metastasis

Objective

To determine if the density of FOXP3⁺ lymphocytes in primary tumors and lymph nodes in pancreatic cancer correlates with the presence of lymph node metastases.

Methods

FOXP3⁺ lymphocyte density in primary pancreatic cancer tissue and draining lymph nodes was measured using immunohistochemistry. We analyzed the clinical and pathological aspects associated with the accumulation of FOXP3⁺ lymphocytes in pancreatic cancer. We also analyzed the correlation of density of FOXP3⁺ lymphocytes in lymph nodes with the nodal status and distance from the primary tumor.

Results

FOXP3⁺ lymphocyte density in pancreatic cancer was significantly higher than in paratumoral pancreatic tissue. The density of FOXP3⁺ lymphocytes in local tumor tissue correlated significantly with the histological grade and overall lymph node status. Furthermore, FOXP3⁺ lymphocyte density was significantly higher in positive lymph nodes than in negative ones, while it had no correlation with the distance of the lymph node from the primary tumor.

Conclusion

FOXP3⁺ lymphocyte density in primary tumor tissue in patients with pancreatic cancer correlates with lymph node metastasis. Lymph nodes containing metastases having higher FOXP3⁺ lymphocyte densities than do negative lymph nodes.     

Lymphocyte maintenance during healthy aging requires no substantial alterations in cellular turnover

In healthy humans, lymphocyte populations are maintained at a relatively constant size throughout life, reflecting a balance between lymphocyte production and loss. Given the profound immunological changes that occur during healthy aging, including a significant decline in T‐cell production by the thymus, lymphocyte maintenance in the elderly is generally thought to require homeostatic alterations in lymphocyte dynamics. Surprisingly, using in vivo ²H₂O labeling, we find similar dynamics of most lymphocyte subsets between young adult and elderly healthy individuals. As the contribution of thymic output to T‐cell production is only minor from young adulthood onward, compensatory increases in peripheral T‐cell division rates are not required to maintain the T‐cell pool, despite a tenfold decline in thymic output. These fundamental insights will aid the interpretation of further research into aging and clinical conditions related to disturbed lymphocyte dynamics.     

A close relationship between cytotoxic T lymphocytes generated in the mixed lymphocyte culture and insulin receptor-bearing lymphocytes: enrichment by density gradient centrifugation.

In contrast to liver, fat, muscle, the fibroblast, and the monocyte, the lymphocyte does not bear insulin receptors unless it is activated by antigen or mitogen. Antigen stimulation by skin graft or in the mixed lymphocyte culture (MLC) generates a population of T lymphocytes, the effector function of which can be augmented by insulin. In the same pool of cells are found T lymphocytes with newly emergent insulin receptors capable of supporting this augmentation. This study demonstrates the close relationship between the augmentable effector T cell and the insulin receptor-bearing cell and strongly suggests that these cells are identical. Splenic lymphocytes from unidirectional murine MLCs were separated into light and heavy fractions by discrete density gradient eentrifugation daily and assayed for cellular-mediated cytotoxicity and for insulin receptors. Receptor-bearing and cytotoxic lymphocytes waxed and waned together primarily in the light fraction. Receptor-positive cells preceded effectors by 24 hr and the two characteristics were highly correlated over time (r ≥ 0.95). T-Cell depletion by specific antisera or by immunoabsorbent column chromatography demonstrated that most, but not all, receptor-bearing cells were T cells and that virtually all effectors were also receptor positive. When the insulin receptor was functionally removed from the lymphocyte membrane by trypsin proteolysis, effector function ceased. The return of cytotoxicity was accompanied by return of the lymphocyte insulin receptor. Receptor-bearing cells were predominantly of the Ly 2⁺3⁺ pedigree but Ly 1⁺ cells were also induced to bear the insulin receptor along with a few non-T cells. These data show that the emergence of a lymphocyte insulin receptor is not just a fortuitous marker event of cellular activation but provides a structure capable of supporting lymphocyte effector function. The appearance of Ly 1⁺ receptor-bearing cells suggests the alloactivation of T helpers and their participation in a T-T cooperative event.     

Antibody-dependent cellular cytotoxicity in the rat, rabbit, and guinea pig: relationship of killer (K) cell activity with the presence of Fc+ C3- and Fc+ C3+ lymphocytes.

ADCC of lymphocyte suspensions from the spleen, lymph node, and blood in the rat, rabbit, and guinea pig, was directly correlated with the presence of Fc⁺ C₃^? and indirectly with the presence of Fc⁺ C₃⁺ lymphocyte subpopulations in these preparations. Rabbit lymphocytes, from all three lymphoid compartments, were deficient in the Fc⁺ C₃^? subpopulation but enriched in the Fc⁺ C₃⁺ subpopulation and lacked K-cell activity. A similar lymphocyte profile and lack of K-cell activity was found in the lymph node of the rat and guinea pig. Lymphocytes from the blood and spleen, on the other hand, had a substantial population of Fc⁺ C₃^? lymphocytes and a prominent K-cell activity. The Fc⁺ C₃⁺ lymphocyte, although unable to lyse IgG-coated target cells, is able to compete in vitro with the Fc⁺ C₃^? (K) cell and significantly reduce ADCC. Phagocytic cells lyse very effectively erythroid target cells and contamination by such cells may explain previously reported K-cell activity of rabbit lymphocytes.     

The involvement of cAMP in lymphocyte capping

We have observed that agents that are known to elevate intracellular levels of cAMP such as N⁶,O²-dibutyryl adenosine 3′,5′-cyclic monophosphoric acid (dbcAMP) and theophylline cause a remarkable stimulatory effect on the lymphocyte receptor mobility phenomenon. Increased intracellular concentration of cAMP enhances not only antibody-induced but also Con A-induced lymphocyte capping events in T-lymphoma cells. In addition, we have noted that N²,O²-dibutyryl guanosine 3′,5′-cyclic monophosphoric acid (dbcGMP) does not stimulate but actually slightly inhibits the receptor movement. Furthermore, we have determined cAMP levels to increase greater than twofold during ligand-induced capping using a radioimmunoassay. Therefore, our data strongly suggest that cyclic adenylic monophosphoric acid (and not cGMP) is specifically involved in the redistribution of lymphocyte membrane proteins induced by both antibody and Con A.     

Human T Lymphocyte Isolation,Culture and Analysis of Migration In Vitro

The migration of T lymphocytes involves the adhesive interaction of cell surface integrins with ligands expressed on other cells or with extracellular matrix proteins. The precise spatiotemporal activation of integrins from a low affinity state to a high affinity state at the cell leading edge is important for T lymphocyte migration ¹. Likewise, retraction of the cell trailing edge, or uropod, is a necessary step in maintaining persistent integrin-dependent T lymphocyte motility ². Many therapeutic approaches to autoimmune or inflammatory diseases target integrins as a means to inhibit the excessive recruitment and migration of leukocytes ³. To study the molecular events that regulate human T lymphocyte migration, we have utilized an in vitro system to analyze cell migration on a two-dimensional substrate that mimics the environment that a T lymphocyte encounters during recruitment from the vasculature. T lymphocytes are first isolated from human donors and are then stimulated and cultured for seven to ten days. During the assay, T lymphocytes are allowed to adhere and migrate on a substrate coated with intercellular adhesion molecule-1 (ICAM-1), a ligand for integrin LFA-1, and stromal cell-derived factor-1 (SDF-1). Our data show that T lymphocytes exhibit a migratory velocity of ~15 μm/min. T lymphocyte migration can be inhibited by integrin blockade ¹ or by inhibitors of the cellular actomyosin machinery that regulates cell migration ².Download video file.^{(98M, mp4)}     

Mst1 controls lymphocyte trafficking and interstitial motility within lymph nodes

The regulation of lymphocyte adhesion and migration plays crucial roles in lymphocyte trafficking during immunosurveillance. However, our understanding of the intracellular signalling that regulates these processes is still limited. Here, we show that the Ste20-like kinase Mst1 plays crucial roles in lymphocyte trafficking in vivo. Mst1^−/− lymphocytes exhibited an impairment of firm adhesion to high endothelial venules, resulting in an inefficient homing capacity. In vitro lymphocyte adhesion cascade assays under physiological shear flow revealed that the stopping time of Mst1^−/− lymphocytes on endothelium was markedly reduced, whereas their L-selectin-dependent rolling/tethering and transition to LFA-1-mediated arrest were not affected. Mst1^−/− lymphocytes were also defective in the stabilization of adhesion through α4 integrins. Consequently, Mst1^−/− mice had hypotrophic peripheral lymphoid tissues and reduced marginal zone B cells and dendritic cells in the spleen, and defective emigration of single positive thymocytes. Furthermore, Mst1^−/− lymphocytes had impaired motility over lymph node-derived stromal cells and within lymph nodes. Thus, our data indicate that Mst1 is a key enzyme involved in lymphocyte entry and interstitial migration.     

Chicken Harder's gland: evidence for a relatively pure bursa-dependent lymphoid cell population

Theophylline, caffeine, and dibutyryl cAMP, agents that elevate intracellular levels of cyclic AMP, were found to inhibit cytotoxin elaboration by PHA-stimulated human lymphocytes. Cytotoxins in diluted supernatants from 3-day lymphocyte cultures were assayed by ⁵¹Cr release from L cells. Addition of the agents to the lymphocyte cultures inhibited cytotoxin elaboration 70–90% at 10^?3M and 20–50% at 10^?5M. In addition, it was found that the inhibition is reversible and occurs at a step other than the initial mitogen triggering.The inhibition of cytotoxin elaboration by these agents correlates strikingly with their inhibition of lymphocyte-mediated target cell lysis. The results are consistent with the hypothesis that cytotoxin is the mediator of target cell killing by lymphocytes.