scholarly journals Sequential responses of mouse spleen T cells in mixed lymphocyte culture-induced cytolysis.

1975 ◽  
Vol 141 (2) ◽  
pp. 508-512 ◽  
Author(s):  
P Häyry ◽  
L C Andersson
Keyword(s):  
T Cells ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Mouse Spleen ◽  
Blast Transformation

T cells triggered to blast transformation and proliferation by histoincompatible cells have the capacity of reverting "back" to lymphocytes. These "secondary" lymphocytes and their progeny cells are able to respond repeatedly to the same allogeneic stimulus in vitro.

scholarly journals Haplotype-specific suppression of cytotoxic T cell induction by antigen inappropriately presented on T cells.

1983 ◽  
Vol 157 (1) ◽  
pp. 141-154 ◽  
Author(s):  
P J Fink ◽  
I L Weissman ◽  
M J Bevan
Keyword(s):  
T Cells ◽  
Cytotoxic T Lymphocyte ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Cytotoxic T Cell ◽  
Spleen Cells ◽  
Specific Suppression ◽  
Veto Cells

To detect a strong cytotoxic T lymphocyte (CTL) response to minor histocompatibility (H) antigens in a 5-d mixed lymphocyte culture, it is necessary to use a responder that has been primed in vivo with antigen-bearing cells. It has previously been shown that minor-H-specific CTL can be primed in vivo both directly by foreign spleen cells and by presentation of foreign minor H antigens on host antigen-presenting cells. This latter route is evident in the phenomenon of cross-priming, in which H-2 heterozygous (A x B)F1 mice injected 2 wk previously with minor H-different H-2A (A') spleen cells generate both H-2A- and H-2B-restricted minor-H-specific CTL. In a study of the kinetics of direct- vs. cross-priming to minors in F1 mice, we have found that minor H-different T cells actually suppress the induction of virgin CTL capable of recognizing them. CTL activity measured from F1 mice 3-6 d after injection with viable A' spleen cells is largely H-2B restricted. The H-2A-restricted response recovers such that roughly equal A- and B-restricted activity is detected in mice as early as 8-10 d postinjection. This temporary hyporeactivity does not result from generalized immunosuppression--it is specific for those CTL that recognize the foreign minor H antigen in the context of the H-2 antigens on the injected spleen cells. The injected spleen cells that mediate this suppression are radiosensitive T cells; Lyt-2+ T cells are highly efficient at suppressing the induction of CTL in vivo. No graft vs. host reaction by the injected T cells appears to be required, as suppression of direct primed CTL can be mediated by spleen cells that are wholly tolerant of both host H-2 and minor H antigens. Suppression cannot be demonstrated by in vitro mixing experiments. Several possible mechanisms for haplotype-specific suppression are discussed, including inactivation of responding CTL by veto cells and in vivo sequestration of responding CTL by the injected spleen cells.

scholarly journals Phenotype, Localization, and Mechanism of Suppression of CD4+CD25+ Human Thymocytes

2002 ◽  
Vol 196 (3) ◽  
pp. 379-387 ◽  
Author(s):  
Francesco Annunziato ◽  
Lorenzo Cosmi ◽  
Francesco Liotta ◽  
Elena Lazzeri ◽  
Roberto Manetti ◽  
...  
Keyword(s):  
T Cells ◽  
Transforming Growth Factor ◽  
Mixed Lymphocyte Culture ◽  
Interferon Γ ◽  
Lymphocyte Culture ◽  
Target Cells ◽  
Suppressive Activity ◽  
Α Chain ◽  
Tgf Β1

Phenotypic markers, localization, functional activities, and mechanisms of action in vitro of CD4+CD25+ T cells, purified from postnatal human thymuses, were investigated. These cells showed poor or no proliferation in mixed lymphocyte culture (MLC), and suppressed in a dose-dependent fashion the proliferative response to allogeneic stimulation of CD4+CD25− thymocytes. Virtually all CD4+CD25+ thymocytes constitutively expressed cytoplasmic T lymphocyte antigen (CTLA)-4, surface tumor necrosis factor type 2 receptor (TNFR2), and CCR8. They prevalently localized to perivascular areas of fibrous septa and responded to the chemoattractant activity of CCL1/I-309, which was found to be produced by either thymic medullary macrophages or fibrous septa epithelial cells. After polyclonal activation, CD4+CD25+ thymocytes did not produce the cytokines interleukin (IL)-2, IL-4, IL-5, IL-13, interferon γ, and only a very few produced IL-10, but all they expressed on their surface CTLA-4 and the majority of them also transforming growth factor (TGF)-β1. The suppressive activity of these cells was contact dependent and associated with the lack of IL-2 receptor (IL-2R) α-chain (CD25) expression in target cells. Such a suppressive activity was partially inhibited by either anti–CTLA-4 or anti–TGF-β1, and was completely blocked by a mixture of these monoclonal antibodies, which were also able to restore in target T cells the expression of IL-2R α-chain and, therefore, their responsiveness to IL-2. These data demonstrate that CD4+CD25+ human thymocytes represent a population of regulatory cells that migrate in response to the chemokine CCL1/I-309 and exert their suppressive function via the inhibition of IL-2R α-chain in target T cells, induced by the combined activity of CTLA-4 and membrane TGF-β1.

scholarly journals Effect of activated lymphocytes on the regulation of hematopoiesis: enhancement and suppression of in vitro BFU-E growth by T cells stimulated by autologous non-T cells

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1143-1147 ◽  
Author(s):  
M Harada ◽  
S Nakao ◽  
K Kondo ◽  
K Odaka ◽  
M Ueda ◽  
...  
Keyword(s):  
T Cells ◽  
Mononuclear Cells ◽  
Colony Growth ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
T Cell Subsets ◽  
Activated T Cells ◽  
Peripheral Blood Mononuclear ◽  
Erythroid Progenitor

Abstract Autologous mixed lymphocyte culture (AMLR) is an immunologic response with memory and specificity and plays a role in immune regulation. Effects of T cells activated by AMLR were studied in the regulation of in vitro erythropoiesis. AMLR-activated T cells were cocultured with autologous non-T, nonphagocytic peripheral blood mononuclear cells for assaying erythroid progenitor cells (BFU-E). T cells activated for 3 days in AMLR showed significant enhancement of in vitro colony growth by BFU-E. In contrast, activated T cells from day 7 AMLR caused significant suppression of BFU-E growth. Both enhancing and suppressing activities of AMLR-activated T cells were mediated by an la-positive and radiosensitive population within the OKT4+ subset. These observations suggest that AMLR-activated T cells may play a role in the immune-mediated regulation of in vitro erythropoiesis. It is also suggested that heterogeneous T-cell subsets may exert regulatory functions in the regulation of in vitro hematopoiesis.

scholarly journals Effect of activated lymphocytes on the regulation of hematopoiesis: enhancement and suppression of in vitro BFU-E growth by T cells stimulated by autologous non-T cells

Blood ◽  
1986 ◽  
Vol 67 (4) ◽  
pp. 1143-1147 ◽  
Author(s):  
M Harada ◽  
S Nakao ◽  
K Kondo ◽  
K Odaka ◽  
M Ueda ◽  
...  
Keyword(s):  
T Cells ◽  
Mononuclear Cells ◽  
Colony Growth ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
T Cell Subsets ◽  
Activated T Cells ◽  
Peripheral Blood Mononuclear ◽  
Erythroid Progenitor

Autologous mixed lymphocyte culture (AMLR) is an immunologic response with memory and specificity and plays a role in immune regulation. Effects of T cells activated by AMLR were studied in the regulation of in vitro erythropoiesis. AMLR-activated T cells were cocultured with autologous non-T, nonphagocytic peripheral blood mononuclear cells for assaying erythroid progenitor cells (BFU-E). T cells activated for 3 days in AMLR showed significant enhancement of in vitro colony growth by BFU-E. In contrast, activated T cells from day 7 AMLR caused significant suppression of BFU-E growth. Both enhancing and suppressing activities of AMLR-activated T cells were mediated by an la-positive and radiosensitive population within the OKT4+ subset. These observations suggest that AMLR-activated T cells may play a role in the immune-mediated regulation of in vitro erythropoiesis. It is also suggested that heterogeneous T-cell subsets may exert regulatory functions in the regulation of in vitro hematopoiesis.

scholarly journals Continuously proliferating allospecific T cells. I. Specificity of cooperation with allogeneic B cells in the humoral antibody response to sheep erythrocytes.

1979 ◽  
Vol 149 (4) ◽  
pp. 808-814 ◽  
Author(s):  
J D Waterfield ◽  
G Dennert ◽  
S L Swain ◽  
R W Dutton
Keyword(s):  
T Cells ◽  
B Cells ◽  
Humoral Response ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Strain Specificity ◽  
Sheep Erythrocytes ◽  
Effect Factor ◽  
Helper Activity

Allospecific mouse T cells, positively selected in one-way mixed lymphocyte culture were maintained for 3 yr in tissue culture by sequential restimulation. Such proliferating T cells were tested for their ability to induce a positive allogeneic effect: activating B cells in an in vitro primary humoral response to sheep erythrocytes. It was found that such T lymphocytes could function as helper cells. Helper activity was shown to be specific in that the B cells activated had to share major histocompatibility complex (H-2) antigens with the strain used for selection of the cell line. Intra H-2 mapping showed that antigens coded in the IAk subregion played an important role in the induction of the positive allogeneic effect. Supernatant factors could substitute for the allogeneic T cells in activation of the in vitro humoral response. However, such supernates exhibited no strain specificity. Therefore, the specificity seen in the positive allogeneic effect is presumably a consequence of the alloantigenic recognition receptors intrinsic to the T cells, and not to any biologically restricting properties of the allogeneic effect factor itself.

scholarly journals Glycolytic Inhibitor 3-Brpa Inhibits Proliferation and Induces Apoptosis of Mouse Splenic Lymphocytes in Mixed Lymphocytes Culture

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4975-4975
Author(s):  
Ruiqing Zhou ◽  
Huiqing He ◽  
Ziwen Guo ◽  
Dafa Qiu ◽  
Weihua Li ◽  
...  
Keyword(s):  
T Cells ◽  
Research Funding ◽  
Inhibitory Effect ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Mouse Spleen ◽  
Control Group ◽  
Spleen Cells ◽  
Guangzhou City ◽  
Ifn Γ

Abstract Background: T-cell activation plays a critical role in the pathogenesis of acute graft-versus-host disease (GVHD). Quiescent T cells utilize oxidative phosphorylation to generate ATP, whereas activated T cells utilize glycolysis, so use glycolysis inhibitor may be a metabolically regulator needed to control T cells induced GVHD. The mixed lymphocytes culture (MLC) was used as a model to evaluate the effect of treatment for GVHD in vitro. Glucolysis inhibitor 3-Bromopyruvic acid (3-BrPA), a glucolysis inhibitor, can effectively induce multidrug resistance leukemia cell lines apoptosis and enhanced chemotherapy-induced cytotoxity to leukemia cells. Objective : This study aimed to study the effects of glycolytic inhibitor 3-Bromopyruvate (3-BrPA) on the proliferation, the apoptosis, the T lymphocyte subsets and the contents of cytokine IL-4 and IFN-γ in mouse spleen cells harvested from mixed lymphocyte culture. Methods An one-way mixed lymphocyte culture system characterized by labeled responder cells with BALB/c mouse spleen cells (H-2kd) and stimulator cells with C57BL/6 mouse spleen cells (H-2kb) was established. With treatment of 3-BrPA at different concentrations (0-200 μmol/L), the CCK-8 method was applied for lymphoproliferation activity, flow cytometry for cell surface markers of CD3, CD4 and CD8, and ELISA method for the levels of cytokine IL-4 and IFN-γ in the supernatant. Results: The CCK-8 test revealed that 3-BrPA in middle or high concentrations (over IC 30, 20 μmol/L) displayed a dose-dependent inhibitory effect on T-cell proliferation of MLC system. The IC50 were 48.6、41.2 and 41.9 μmol/L after 24 h, 36 h and 48 h of culture, respectively. FCM test discovered that the inhibitory effect mainly occurred in the CD4+ cells. After 48 h of culture, the apoptosis rate of 0, 10, 20, 50 and 100 μmol/L group were 4.86±0.88%, 5.2±1.13%, 12.63±2.97%, 18.55±4.06% and 22.47±3.61%, respectively. With treatment of 20 or 50μmol/L 3-BrPA, the levels of IFN-γ decreased obviously to 243.37±15.64 ng/L and 164.25±20.14 ng/L, compared with the control group (277.61±18.46 ng/L). The levels of IL-4 increased mildly to 33.18±5.69 ng/L and 31.06±6.06 ng/L, compared with the control group (28.64±3.97ng/L). Thus, the IFN-γ/IL-4 ratio decreased significantly. Conclusions :The results indicated that 3-BrPA could inhibit T cells proliferation, induce apoptosis and contribute to the Th2 cytokine environment in murine mixed lymphocyte culture system. Disclosures Liu: National Natural Science Foundation of China (81270647, 81300445, 81200388): Research Funding; National High Technology Research and Development Program of China (863 Program) (2011AA020105): Research Funding; National Public Health Grand Research Foundation (201202017): Research Funding; Natural Science Foundation of Guangdong Province (S2012010009299): Research Funding; the project of health collaborative innovation of Guangzhou city (201400000003-4, 201400000003-1): Research Funding; the Technology Plan of Guangdong Province of China (2012B031800403): Research Funding; the project of the Zhujiang Science & Technology Star of Guangzhou city (2013027): Research Funding.

ONTOGENY OF IMMUNITY IN MAN

1975 ◽  
Vol 80 (2_Suppl) ◽  
pp. S306-S317 ◽  
Author(s):  
Daniel P. Stites ◽  
Joseph Caldwell ◽  
Martin C. Carr ◽  
H. Hugh Fudenberg
Keyword(s):  
T Cells ◽  
Human Subjects ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Lymphoid Tissues ◽  
Foetal Development ◽  
Lymphoid Development ◽  
Foetal Liver ◽  
Low Levels

ABSTRACT We have reviewed selected aspects of recent findings in the ontogeny of immunity in man. For obvious reasons, constraints placed on experimental work with human subjects, albeit deceased foetuses, limit the work to in vitro studies. Nevertheless, a number of novel and important concepts have emerged. First, as in lower animals, lymphoid development of the foetal thymus in general precedes the development of immunocompetence in peripheral lymphoid tissues. A striking exception to this rule is the finding of cells in early foetal liver which respond to allogeneic cells in the mixed lymphocyte culture some weeks before lymphoid organization of the thymus. In addition, the response of foetal cells to allogeneic cells in the mixed lymphocyte reaction (MLR) precedes the response to phytohaemagglutinin (PHA), a stimulant with relative T cell specificity. The use of markers for T cells to map the emergence of this class of lymphocytes correlates well with various functional attributes of T cells in in vitro culture systems. B cells emerge first in foetal liver at about 9 weeks' gestation, but formation of immunoglobulin by the foetus occurs at very low levels until after birth. There is a suggestion that progression from IgM to IgA synthesis occurs during human foetal lymphoid development. Taken as a whole, these data suggest a rather remarkable and perhaps unexpected degree of cellular and potential humoral immunocompetence at early stages of foetal development in man.

scholarly journals Autologous mixed lymphocyte culture reactions and generation of cytotoxic T cells.

1977 ◽  
Vol 146 (6) ◽  
pp. 1809-1814 ◽  
Author(s):  
R A Vande Stouwe ◽  
H G Kunkel ◽  
J P Halper ◽  
M E Weksler
Keyword(s):  
T Cells ◽  
Cell Lines ◽  
Lymphoid Cell ◽  
Cytotoxic T Cells ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Cell Generation ◽  
Cytotoxic T Cell ◽  
Cytotoxic Cells ◽  
Lymphoid Cell Lines

Autologous mixed lymphocyte culture (MLC) reactions were studied utilizing autologous purified B cells and autologous established B lymphoid cell lines as stimulating cells. Similar results were obtained although somewhat greater stimulation of lymphocyte proliferation was found with the autologous lymphoid cell lines. Cytotoxic T cells were not generated against the stimulating cells in either case when peripheral blood cells were used as targets. A low cytotoxicity was detected when lymphoid cell lines were used both as stimulators and target cells. However this was nonspecific and was always greater for heterologous lines than for the stimulator line. Third-party cell experiments demonstrated that the autologous reaction could serve as a proliferative stimulus for specific cytotoxic lymphocyte generation. Heat-treated allogeneic lymphocytes that alone do not stimulate proliferation ro cytotoxic T-cell generation in MLC reactions when added to the autologous system produced specific cytotoxic cells. The separation of the proliferative phase from the cytotoxic cell generation was especially striking in these experiments. Possible uses of this system for the generation of specific cytotoxic cells to other nonstimulatory cells are discussed.

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