scholarly journals ROLE OF H-2 LYMPHOCYTE-DEFINED AND SEROLOGICALLY-DEFINED COMPONENTS IN THE GENERATION OF CYTOTOXIC LYMPHOCYTES

1974 ◽  
Vol 140 (5) ◽  
pp. 1410-1415 ◽  
Author(s):  
Barbara J. Alter ◽  
Fritz H. Bach
Keyword(s):  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Mouse Spleen ◽  
Target Cells ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Efficient Generation

The cell-mediated lympholytic capability of mouse spleen cells stimulated in mixed lymphocyte culture is determined by lymphocyte-defined (LD) and serologically-defined (SD) antigenic differences present during sensitization. Cells which are activated by LD differences alone are markedly less effective in causing lysis of target cells. This lack of cytotoxicity is shown to be, at least in part, due to the inability of LD differences to allow the efficient generation of cytotoxic lymphocytes. SD antigens not only serve as good targets for CML but are also shown to be important for the generation of cytotoxic lymphocytes during the mixed lymphocyte culture.

scholarly journals SYNERGY BETWEEN SUBPOPULATIONS OF MOUSE SPLEEN CELLS IN THE IN VITRO GENERATION OF CELL-MEDIATED CYTOTOXICITY

1974 ◽  
Vol 140 (6) ◽  
pp. 1646-1659 ◽  
Author(s):  
Richard J. Hodes ◽  
Barry S. Handwerger ◽  
William D. Terry
Keyword(s):  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Specific Cell ◽  
Spleen Cells ◽  
Normal Spleen ◽  
In Vitro Induction ◽  
Cytotoxic Effector ◽  
Nylon Wool

Two subpopulations separated from normal spleen have been shown to synergize as responding cells in the in vitro induction of specific cell-mediated cytotoxicity during the mixed lymphocyte culture (MLC). The synergizing populations are a nylon wool column-adherent and a nylon wool column-nonadherent fraction, enriched for B lymphocytes and T lymphocytes, respectively. When a mixture of these fractions is used as the responding cell population in MLC, greater cytotoxicity is generated than would be expected from the sum of activities generated in the two subpopulations sensitized separately. The synergy appears to occur at the sensitization rather than the effector phase. The synergizing cell which is contained in the nylon-adherent subpopulation is distinct from the cytotoxic effector T lymphocyte, is resistant to lysis by rabbit antimouse brain serum, and is unresponsive to phytohemagglutinin; its synergizing function could not be replaced by either plastic-adherent spleen cells or peritoneal exudate cells. These results suggest a role of a non-T-cell nonmacrophage population in the generation of cytotoxic activity.

scholarly journals CELL-MEDIATED LYMPHOLYSIS

1973 ◽  
Vol 137 (5) ◽  
pp. 1303-1309 ◽  
Author(s):  
Barbara J. Alter ◽  
Dolores J. Schendel ◽  
Marilyn L. Bach ◽  
Fritz H. Bach ◽  
Jan Klein ◽  
...  
Keyword(s):  
Major Histocompatibility Complex ◽  
Target Cell ◽  
Effector Cell ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Mouse Spleen ◽  
Spleen Cells ◽  
Histocompatibility Complex ◽  
Region Difference ◽  
Mediate Cell

The cell-mediated lympholytic capability of mouse spleen cells stimulated in mixed lymphocyte culture is related to the major histocompatibility complex genotype on target lymphocytes. The strain combinations AQR-B10. T(6R) and B10.A(4R)-B10.A(2R) that result in significant mixed lymphocyte culture activation do not mediate cell-mediated lympholysis on sensitizing target lymphocytes; serologically defined regions (H-2K and H-2D) are identical within each combination. H-2K or H-2D region disparity alone does not cause cell-mediated lympholysis. However after mixed lymphocyte culture activation as seen with B10.A-B10.T(6R), a target cell bearing only an H-2K region difference from the effector cell is sensitive to cell-mediated lympholysis. Likewise an H-2D region difference is an adequate target after mixed lymphocyte culture activation of the effector cell in the combination B10.A(2R)-B10.D2.

scholarly journals The effect of allogeneic presensitization on H-Y graft survival and in vitro cell-mediated responses to H-y antigen.

1976 ◽  
Vol 144 (3) ◽  
pp. 810-820 ◽  
Author(s):  
R D Gordon ◽  
B J Mathieson ◽  
L E Samelson ◽  
E A Boyse ◽  
E Simpson
Keyword(s):  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
The Other ◽  
Target Cells ◽  
Spleen Cells ◽  
Cytotoxic Cells ◽  
Parental Haplotype ◽  
Cytotoxic Responses

C57BL/6 and C57BL/10 female mice were grafted with skin from male or female donors incompatible for H-2 and/or non-H-2 antigens. Syngeneic male grafts applied after the rejection of primary allografts or syngeneic male grafts were rejected in accelerated (second set) fashion, whereas male grafts applied after primary female grafts were not. In addition, C57BL/10 female spleen cells, primed in vivo with an allogeneic (BALB/c, CBA, or B10.BR) male graft and challenged in vitro in mixed lymphocyte culture with syngeneic (C57BL/10) male cells, produced cytotoxic cells specific for syngeneic male target cells. We conclude that at least some component of H-Y is detected by female responder cells on allogeneic male cells, and that the second set cell mediated response to H-Y is not necessarily restricted by the H-2 haplotype of the primary sensitizing strain. Moreover, (CBA X B10) F1 females, primed in vivo with male cells of one parental haplotype (B10 or CBA) and challenged in vitro with male cells of the other parental haplotype (CBA or B10), fail to lyse male target cells of either parental haplotype. It therefore seems unlikely that a helper determinant shared between B10 and CBA is sufficient to explain the ability of CBA male cells to prime H-2-restricted T-cell cytotoxic responses by B10 females.

scholarly journals Concanavalin A potentiates syngeneic response in murine lymphocytes.

1976 ◽  
Vol 143 (1) ◽  
pp. 1-14 ◽  
Author(s):  
K Ozato ◽  
J D Ebert
Keyword(s):  
Concanavalin A ◽  
Proliferative Response ◽  
Specific Inhibitor ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Con A ◽  
Strong Suppression ◽  
Two Factors

In an attempt to modulate the recognition processes that occur on lymphocyte membranes in mixed lymphocyte culture, responding cortisone resistant thymocytes or stimulating spleen cells (treated with mitomycin C) were pretreated with native concanavalin A (N-Con A) or succinyl-Con A (S-Con A). Highly significant cell proliferation was observed in syngeneic combinations when either the responding cells or the stimulating cells were so treated with Con A, although Con A pretreatment alone was never mitogenic. In allogeneic combinations the proliferative response with Con A pretreatment of either partner on day 3 was five to seven times higher than in the normal mixed lymphocyte reactions. The triggering of proliferation was dependent on two factors: (a) The presence of spleen cells as the stimulating cells (thymocytes were much less effective). (b) The binding of Con A molecules to either one of the partners, the effect being abrogated by the specific inhibitor of Con A, alpha-mannopyranoside. The optimal concentration of S-Con A was about twice that of N-Con A. Even more striking was the observation that cultures in which either one of the partners was pretreated with Con A in allogeneic combinations showed a strong suppression (60-80% inhibition) in the subsequent generation of the cytotoxic lymphocytes (CL). The Con A concentration required to trigger a proliferative response corresponded to that for suppressing the generation of CL. Con A pretreatment did not result in a cytotoxic activity toward syngeneic tumor cells.

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.

scholarly journals Primary in vitro cytotoxic response of NZB spleen cells to Qa-1b-associated antigenic determinants.

1979 ◽  
Vol 150 (6) ◽  
pp. 1555-1560 ◽  
Author(s):  
R R Rich ◽  
D A Sedberry ◽  
D L Kastner ◽  
L Chu
Keyword(s):  
Primary Cultures ◽  
Antigenic Determinants ◽  
Target Cells ◽  
Cytotoxic Lymphocytes ◽  
Spleen Cells ◽  
Histocompatibility Antigens ◽  
Viral Antigens ◽  
In Vitro Response

We have shown that cytotoxic lymphocytes generated in primary cultures of NZB spleen cells with H-2-identical BALB/c or B10.D2 stimulator cells exhibit specificity for Qa-1b-associated antigenic determinants. This unidirectional cytotoxicity constitutes the initial demonstration of a primary in vitro response to antigens of the Qa-Tla system. Such responses do not require H-2 homology between effector and target cells in the assay system. In fact, when H-2Dd homologous target cells were employed there was little, if any, evidence for development of primary H-2-restricted responses to minor locus histocompatibility antigens or viral antigens. In view of the recently defined role of Qa-1+, Ly-1,2,3+ cells as regulators of antibody responses, and of the deficiency of such cells in NZB mice, the observation of hyperreactivity for determinants of this system may be relevant to the development of autoimmunity in these animals.

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.

Primary Immune Response of Mouse Spleen Cells in a Serum-Free System: the Role of 2-Mercaptoethanol

Pathobiology ◽  
1980 ◽  
Vol 48 (1) ◽  
pp. 45-50
Author(s):  
M. Burger ◽  
M.W. Hess ◽  
H. Cottier
Keyword(s):  
Immune Response ◽  
Primary Immune Response ◽  
Mouse Spleen ◽  
Spleen Cells ◽  
Free System ◽  
Serum Free

scholarly journals Study of the cells proliferating in parent versus F hybrid mixed lymphocyte culture.

1975 ◽  
Vol 141 (4) ◽  
pp. 775-787 ◽  
Author(s):  
P F Piguet ◽  
H K Dewey ◽  
P Vassalli
Keyword(s):  
Cell Proliferation ◽  
T Cells ◽  
T Lymphocytes ◽  
Hybrid Cell ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Lymphoid Cells ◽  
Spleen Cells ◽  
F1 Hybrid ◽  
Proliferation And Differentiation

Caryotypic analysis of the cells dividing in mouse parent-hybrid MLC showed an F1 hybrid cell proliferation, which varied depending upon the source of lymphoid cells used: strong in spleen MLC (sometimes equal to that of the parental cells), less marked in lymph node cell MLC, and most often absent in MLC between cortisone-resistant (CR) thymocytes. MLC between parental spleen cells and F1 CR thymocytes showed, however, that in certain conditions of culture F thymocytes can also proliferate. Using parental or F1 spleen cells lacking T lymphocytes, it was found that F1 cell proliferation is entirely dependent upon the presence of parental T cells, but does not require the presence of T lymphocytes among the F1 cells. Immunofluorescence analysis of the blasts observed in one-way MLC showed that about 70% of the parental blasts were T blasts, and 25%B blasts (containing a high proportion of plasmablasts); among the F1 blasts, there was also the same percentage of B blasts and plasmablasts, but many of the T blasts bore only small amounts of T-cell antigen (MTLA), and there was also about 20%of unstained blasts, possibly T blasts bearing MTLA in amounts undetectable by immunofluorescence. The possibility is discussed that the F1 responding T cells belong to a subpopulation performing a suppressive function; MLC lacking F1 T cells showed increased [3H] thymidine incorporation. The proliferation and differentiation of parental and F1 B cells may result mainly from an unspecific, "polyclonal" triggering.

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