Induction of allospecific suppressor T Cells in vitro

doi:10.1016/0008-8749(77)90234-9

Cellular Immunology

Volume 34, Issue 1, November 1977, Pages 112-124

https://doi.org/10.1016/0008-8749(77)90234-9 Get rights and content

Abstract

Incubation of mouse thymic lymphocytes with irradiated allogeneic spleen cells gave rise to suppressor cells. The suppressor activity was assayed by adding the incubated cell mixture to a mixed lymphocyte culture (MLC) in which the responder cells were syngeneic with the sensitized thymocytes and the stimulator cells were syngeneic with the sensitizing spleen cells. Such addition suppressed significantly thymidine incorporation in the mixed lymphocyte reaction (MLR). The suppressor cells were found to carry the θ antigen and to function allospecifically, as shown by cross-testing in three allogeneic combinations. Our data suggest that these cells may originate from immature cortisone-sensitive thymic lymphocytes and also provide some preliminary information concerning their mode of action.

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Cited by (20)

Immunoregulation by mouse T-cell clones. I. Suppression and amplification of cytotoxic responses by cloned H-Y-specific cytolytic T lymphocytes
1984, Cellular Immunology
H-Y-specific and H-2D^b-restricted, Lyt-1⁻2⁺ T-cell clones (CTLL) with graded specific cytotoxic activities on male C57BL/6 (B6) target cells (1E3, +++; 2C5, ++; 2A5, +, 3E6, ±) were tested for their capacity to inhibit the generation of H-Y-specific cytotoxic T lymphocytes (CTL) in vitro. Addition of irradiated lymphocytes of CTLL 1E3 and CTLL 3E6 but not those of CTLL 2A5 or CTLL 2C5 abolished the generation of CTL from in vivo primed H-Y-specific precursor cells (CTLP) when added to fresh mixed-lymphocyte cultures (MLC). Exogenous sources of T-cell growth factors (TCGF) did not overcome suppression. Rather the presence of TCGF resulted in a further enhancement of suppressive activities in CTLL 1E3 and 3E6 and the induction of similar activities in cells from CTLL 2A5 and 2C5, which by themselves were not inhibitory. Moreover when added to similar MLC on Day 1 instead of Day 0, only irradiated cells of CTLL 3E6 but not those of the other three CTLL were suppressive. Induction of suppressive activities in H-Y-specific CTLL was independent of the appropriate male stimulator cells since it was also observed in MLC induced by irrelevant antigens (H-2, trinitrophenol). Furthermore at low cell numbers, irradiated lymphocytes from any of the CTLL consistently enhanced CTL activities generated from H-Y-specific CTLP. This augmenting activity, which was not TCGF, could be transferred by soluble mediators present in antigen-sensitized CTLL cultures. Thus, these data indicate (i) that cytotoxic effector cells can function as suppressor cells in the generation of CTL, (ii) that the cytotoxic activity of cloned CTL does not correlate with their capacity to suppress CTL responses, (iii) that the inhibition of CTL responses by CTLL is not due to simple consumption of T-cell growth factors produced in MLC, and (iv) that different CTL clones may interfere with the generation of CTL at different stages of their maturation. Moreover, the experiments suggest an antigen-independent enhancement of suppression by the interaction of CTL with lymphokines. Together with the augmenting activity evoked by cloned CTL the data provide strong evidence for the expression of multiple immunological functions by one particular subset of T cells and suggest that cytotoxic effector cells can differentially regulate the maturation and/or clonal expression of their precursor cells.
Theory and methods for immunization in culture and monoclonal antibody production
1982, Journal of Immunological Methods
Modulation of allogeneic stimulation in man. I. Characterization of an in vitro induced suppressor macrophage population
1981, Human Immunology
Cultured human peripheral blood mononuclear cells suppressed the allogeneicresponse of fresh autologous lymphocytes. This suppressor activity developed gradually over a period of one week. The cells primarily responsible for this effect were enriched by Ficoll density gradient centrifugation. It was found that the suppressor cell is a large, low density nylon wool adherent, radioresistant, phagocytic, and nonspecific esterase positive mononuclear cell. Moreover, these cells did not form E rosettes and were Fc positive. Electron microscopy confirmed that suppressor cells were macrophagelike. Suppressor activity was not due to cytotoxicity, crowding, or steric hindrance by the cultured cells. The suppressor macrophage population did not appear to inhibit the allogeneic response via prostaglandin or arginase release, or interfere with the tritiated thymidine uptake by release of endogenous thymidine. The above system is viewed as an in vitro model of immune regulation by suppressor macrophages, in the context of allogeneic response.
The plaque-forming-cell (PFC) response of human blood lymphocytes. II. PFC response by mixtures of allogeneic lymphocytes cultured with formalin-treated Staphylococci
1981, Cellular Immunology
Formalin-treated Staphylococci (FSA) induce anti-SRBC PFC in cultures of human lymphocytes. Regulation of the PFC response induced by FSA in cultures containing lymphocytes from two allogeneic donors was studied. The PFC response observed in such cocultures could not be predicted from the responses of lymphocytes from the two donors cultured individually. The PFC response of approximately one half the cocultures was less than expected. The remaining cocultures generated more PFC than expected. The depression or augmentation in the PFC response which was observed in cocultures was reproducible when lymphocytes from the same pair of donors were cocultured. Cocultures containing lymphocytes from identical twins generated the expected PFC response. The data suggest that suppressor or helper activity may be generated during a “two-way” allogeneic mixed lymphocyte reaction (MLR). Much less deviation from the expected PFC response was observed during a “one-way” MLR. Anti-Ia antiserum treatment of either donor's lymphocyte population tended to eliminate the deviation from the expected PFC response in coculture. The data suggest that a feedback loop, involving cells from both donors, may be operating in the “two-way” MLR, which leads to the generation of suppressor or helper activity.
Further characterization of suppressor lymphocytes induced by fetal calf serum in murine lymphoid cell cultures: Comparison with in vitro-generated cytotoxic lymphocytes
1979, Cellular Immunology
Nonspecific suppressor cell (SPC) activity has been induced in vitro by preculturing splenocytes from normal mice in the presence of fetal calf serum (FCS) for 3 days or more. In adoptive transfer experiments in vivo, these precultured SPC were shown to reduce the humoral response of mice to SRBC and the cell-mediated cytotoxic (CMC) response to allogeneic tumor cells. In mixing experiments in vitro, using freshly explanted splenocytes, the precultured splenocytes abrogated the generation of specific cytotoxic lymphocytes (CL) in primary mixed lymphocyte-tumor cell cultures (MLTC). By contrast, secondary cytotoxic response was only marginally affected. Supernatants of precultured cells were also inhibitory, although to a lesser degree than whole cells. The induction of suppressive activity was abolished by addition of mitogenic amounts of concanavalin A to the preculturing medium.
By the use of cell fractionation techniques it was found that both specific CL and nonspecific SPC lack an Fc receptor, do not adhere to nylon wool, and cannot be separated from each other by density sedimentation on a discontinuous BSA gradient. However, precursors of SPC and CL differed in their susceptibility to cyclophosphamide, hydrocortisone, and irradiation. The data presented does not exclude the possibility that suppressive activity exerted by FCS-induced SPC is mediated through a cytotoxic effect.
Blocking of T cell activation at the G1A/G1B interface of the cell cycle and prevention of expression of interleukin 2 receptors by alloactivated suppressor T lymphocytes
1988, Transplantation

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^☆: This work was supported by the Thyssen Foundation.

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Induction of allospecific suppressor T Cells in vitro☆

Abstract

Cell. Immunol

Exp. Cell Res

Contemp. Top. Immunobiol

J. Exp. Med

J. Exp. Med

J. Exp. Med

Transplant. Rev

Transplant. Rev

Eur. J. Immunol

J. Immunol

J. Exp. Med

J. Immunol