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 Cellular basis of neonatal induction of in vitro tolerance.

1976 ◽  
Vol 143 (6) ◽  
pp. 1545-1550 ◽  
Author(s):  
O J Kuperman ◽  
F H Bach
Keyword(s):  
Major Histocompatibility Complex ◽  
T Lymphocytes ◽  
Cytotoxic T Lymphocytes ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Proliferative Phase ◽  
Helper Cells ◽  
Histocompatibility Complex ◽  
Induction Of Tolerance

The LD and SD antigens of the major histocompatibility complex subserve differential roles in the induction of the proliferative phase in mixed lymphocyte culture and in the cytotoxic reaction seen in cell-mediated lympholysis. The present study suggests that they also behave differently in the neonatal induction of tolerance. SD antigens appear to induce tolerance in the cytotoxic T lymphocytes very effectively, whereas LD antigens (or the cytotoxic targets coded by genes in the I and/or S regions) are relatively ineffective in this regard. LD antigens presented neonatally are effective at inducing tolerance in the proliferating helper cells.

scholarly journals Trans-acting element(s) operating across species barriers positively regulate expression of major histocompatibility complex class II genes.

1985 ◽  
Vol 162 (4) ◽  
pp. 1117-1133 ◽  
Author(s):  
R S Accolla ◽  
L Scarpellino ◽  
G Carra ◽  
J Guardiola
Keyword(s):  
Gene Expression ◽  
Major Histocompatibility Complex ◽  
Mhc Class Ii ◽  
Class Ii ◽  
Mouse Spleen ◽  
Chain Gene ◽  
Major Histocompatibility ◽  
Spleen Cells ◽  
Histocompatibility Complex ◽  
Ia Antigens

Raji, a human B lymphoma line, expresses high levels of major histocompatibility complex (MHC) class II antigens. Conversely, none of the detectable human Ia antigens is present in RJ 2.2.5, an immunoselected Raji variant. Clonal analysis, biochemical characterization, and nucleic acid hybridization studies of hybrids between mouse spleen cells and RJ 2.2.5 show that MHC class II gene expression is regulated in trans by a factor which, as judged by dominance studies, has the characteristics of an activator. Such a positive trans acting factor is expressed in mouse spleen cells, and is able to implement MHC class II gene expression across species boundaries. Expression of this factor in spleen cells strongly suggests that it plays a role in in vivo regulation of Ia expression. Additional data suggest that different subsets of class II genes such as DR and DQ may, in part, be regulated by different mechanisms. It has also been possible to show that the amount of In chain-specific mRNA, present at reduced levels in RJ 2.2.5 cells compared to the parental Raji cells, drastically increased in human X mouse cells hybrids reexpressing human Ia antigens, suggesting that the In chain gene and the class II genes, although located on different chromosomes, are regulated in a concerted fashion, either directly through the same implementing factor, or indirectly through a cascade mechanism.

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 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 Mixed lymphocyte culture determinants and C2 deficiency: LD-7a associated with C2 deficiency in four families.

1975 ◽  
Vol 142 (2) ◽  
pp. 495-506 ◽  
Author(s):  
S M Fu ◽  
R Stern ◽  
H G Kunkel ◽  
B Dupont ◽  
J A Hansen ◽  
...  
Keyword(s):  
Major Histocompatibility Complex ◽  
Linkage Disequilibrium ◽  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Major Histocompatibility ◽  
Histocompatibility Complex ◽  
Lymphocyte Cultures ◽  
Tentative Evidence ◽  
Mixed Lymphocyte Cultures ◽  
C2 Deficiency

Four families with C2 deficiency were studied. Among eight HL-A haplotypes involved with C2 deficiency, five were HL-A 10,W18. Three homozygotes for C2 deficiency from different families were mutually nonreactive in mixed lymphocyte cultures (MLC) and the heterozygotes from the fourth family failed to react to the homozygous cells. It appeared that identical MLC determinants were associated with all the genes from the different families that related to C2 deficiency. Further experiments identified the MLC determinant, LD-7a, as being involved. These results suggest marked linkage disequilibrium between the genes for C2 deficiency and the major histocompatibility complex (MHC). Studies of possible recombinants have offered tentative evidence for the positioning of the locus for C2 deficiency with respect to other segments of the MHC.

scholarly journals A MOUSE B-CELL ALLOANTIGEN DETERMINED BY GENE(S) LINKED TO THE MAJOR HISTOCOMPATIBILITY COMPLEX

1973 ◽  
Vol 138 (6) ◽  
pp. 1289-1304 ◽  
Author(s):  
David H. Sachs ◽  
James L. Cone
Keyword(s):  
Major Histocompatibility Complex ◽  
Lymph Node ◽  
T Cell ◽  
B Cells ◽  
Cell Surface ◽  
B Cell ◽  
Surface Antigen ◽  
Major Histocompatibility ◽  
Spleen Cells ◽  
Histocompatibility Complex

Antibodies cytotoxic for only a subpopulation of C57Bl/10 lymph node and spleen cells were detected when rat antiserum against B10.D2 was exhaustively absorbed with B10.A lymphocytes. Antibodies of similar specificity were also detected in B10.A anti-B10.D2 and in B10.A anti-C57Bl/10 alloantisera. Reactions with recombinant strains of mice indicate that the cell-surface antigen(s) responsible for this specificity is determined by gene(s) in or to the left of the Ir-1 region of the major histocompatibility complex. A variety of criteria implicate B cells as the subpopulation of lymphocytes bearing this antigen. In view of these data and the recent report by others of a T-cell alloantigen determined by gene(s) in the major histocompatibility complex, it seems possible that there may be a variety of H-2-linked alloantigens expressed preferentially on subclasses of lymphocytes.

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 Antigen recognition by a T cell clone outside the context of the major histocompatibility complex. A role for Mls in antigen presentation.

1984 ◽  
Vol 159 (1) ◽  
pp. 305-312 ◽  
Author(s):  
S J Waters ◽  
S D Waksal ◽  
G P Norton ◽  
C A Bona
Keyword(s):  
Major Histocompatibility Complex ◽  
T Cell ◽  
Cell Clone ◽  
Antigen Recognition ◽  
Major Histocompatibility ◽  
Spleen Cells ◽  
Differentiation Markers ◽  
Histocompatibility Complex ◽  
T Cell Clone

A T cell clone isolated from antigen-primed CB6/F1 mice was shown to proliferate to keyhole limpet hemocyanin (KLH) in the presence of irradiated syngeneic F1 spleen cells, as well as spleen cells from either parental strain (BALB/c and C57BL/6). The genetic restriction involved in this antigen-specific proliferation was mapped using BXD (C57BL/6 X DBA/2) recombinant inbred strains of mice to the Mls gene on chromosome one. To exclude the role of Ia antigens as the restricting determinants, monoclonal anti-Ia antibodies were used to block the in vitro proliferative response of this clone. Although anti-Iab and anti-Iad blocked the proliferation of this clone to KLH in the presence of irradiated spleen cells from either parent, this effect was shown to be dependent on Ia molecules passively absorbed by the T cell clone from the irradiated filler cells. Since the T clone expressed Thy-1.2 and Lyt-1+ differentiation markers, its helper activity was compared with other KLH carrier-specific clones in an in vitro antibody synthesis assay. The Mls-KLH-restricted T cell clone, in contrast to other carrier-specific, major histocompatibility complex (MHC)-restricted T cell clones, was unable to cooperate with trinitrophenyl (TNP)-primed B cells in the presence of TNP-KLH to generate an anti-TNP response. These experiments suggest that non-MHC determinants, such as autologous Mls gene products, may play a role in genetically restricted antigen recognition by T lymphocytes.

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