Generation of rat TH2-like cells in vitro is interleukin-4-dependent and inhibited by interferon-γ (original) (raw)
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Immunology
Interleukin-4 (IL-4) is antagonistic for many of the activities of interferon-gamma (IFN-gamma) and, as well as suppressing the development of T-helper type-1 (Th1) cells, has been reported to block directly the synthesis of IFN-gamma in human lymphocytes. However, IL-4 transgenic mice produce increased amounts of IFN-gamma as well as IL-4. We have compared the ability of rat IL-4 to regulate IFN-gamma secretion in short-term cultures of spleen cells with its effect on the differentiation of T lymphocytes into IFN-gamma-producing, or Th1-type, cells. Normal rat spleen cells were stimulated using a variety of mitogens and ovalbumin antigen, with or without IL-4, for 12-24 hr and the levels of IFN-gamma in the supernatants measured by enzyme-linked immunosorbent assay (ELISA). The results show that when normal rat splenocytes were stimulated with phytohaemagglutinin (PHA) or concavalin A (Con A), IL-4 enhanced secretion of IFN-gamma after 12-24 hr. This enhancement was also apparent w...
Annals of the New York Academy of Sciences, 1996
Interleukin (1L)-12 and prostaglandin (PG) E, are two immunomodulators produced by antigen-presenting cells (APC) and accessory cells (AC) in response to a variety of stimuli. These immunomodulators exert opposite modulatory effects on cytokine production by activated CD4+ T cells. IL-12 is particularly effective as a selective inducer and enhancer of IFN-y production by T cells' and is an obligatory factor for the generation of T helper type 1 (Thl) PGE, , on the other hand, inhibits IFN-y production by CD4+ T cells in a dose-dependent way, without affecting EL-4 production, resulting in a Th2-like cytokine p r~f i l e .~" Because IL-2 and PGE, are produced by AC and APC and such cells are evidently present in the microenvironment of T cells during T cell activation, the present study focused on the relative contributions of AC-derived IL-12 and PGE, in determining levels of IFN-y, produced by these T cells. First we addressed the question whether the modulatory actions of IL-12 and PGE, mutually interfere by stimulating CD4' T lymphocyte clones with anti-CD3 and anti-CD28 mAb in the presence of increasing concentrations of both IL-12 (1-100 U/ml) and PGE, (10-s-10-5 M). With respect to IFN-y production, IL-12 did not interfere with the downregulatory effects of PGE, ; and, likewise, PGE, did not prevent the IL-12-induced upregulation of IFN-y production. Next, we studied the susceptibility of activated T cells to IL-12 and PGE, in time by adding these modulators at different time points after stimulation. These experiments clearly showed that T cells become insensitive to PGE, within 2 to 6 hours after activation, whereas susceptibility to IL-12 is retained for at least for 8 hours. Together with the crosstitration experiments, these data indicate that the net IFN-y production in these T-cell cultures was determined by the IL-12/PGE2 concentration ratio at the time point of T-cell activation.
European Journal of Immunology, 1990
The production of interleukin 2 (IL 2), IL 4 and interferon-γ (IFN-γ) by in vitro activated unselected human blood mononuclear cells was studied at a single-cell level. Individual lymphokine-synthesizing cells were identified by intracellular immunofluorescent staining using cytokine-specific monoclonal or polyclonal antibodies. Cultures from adult blood donors revealed a biphasic kinetic production pattern for IL 2 and IFN-γ with peaks occurring 4–6 and 24–30 h after initiation of the cultures. Approximately 20%–40% of the lymphocytes produced IL 2 and IFN-γ. In contrast, only 1%–3% of the lymphocytes synthesized IL 4 with maximal frequency after 6 h of culture. CD4+ as well as CD8+ T cells contributed to the synthesis of all three lymphokines studied. CD4+CD45R− T cells were the major producers of IL 2 and IL 4, while CD8+CD45R− T cells were the most common phenotype of IFN-γ-synthesizing cells. By performing two-color immunofluorescence studies we observed that among IL 4-producing cells every second one made simultaneously IL 2 and every fourth one made IFN-γ. Mononuclear cells from umbilical cord blood could be stimulated to make IL 2 to the same extent as cells from adult blood donors. No IL 4 production and a strikingly reduced frequency of IFN-γ producers were noted in cell cultures from neonates. IL 2, IL 4 and IFN-γ accumulated in the Golgi system, which resulted in a characteristic morphology of the staining, eliminating problems with evaluation of background signals.
Simultaneous production of IL-2, IL-4, and IFN-gamma by activated human CD4+ and CD8+ T cell clones
The Journal of Immunology
In the present study, we have investigated the ability of human T cells to secrete IL-2, IL-4, and IFN-gamma. IL-4 and IFN-gamma were quantified with enzymatic immunoassays and IL-2 with a biologic assay by using the murine IL-2-dependent cell line CTLL-2. PBL, stimulated with Con A or with a combination of the phorbol ester 13-O-tetradecanoylphorbol-12-acetate and the Ca2+ ionophore A23187 secreted IL-2, IL-4, and IFN-gamma. The kinetics of the secretion of the three lymphokines was investigated with two CD4+ clones; one (GEO-2) that produced IL-2, IL-4, and IFN-gamma and another (HY640), that produced only IL-2 and IFN-gamma. Significant IL-2, IL-4, and IFN-gamma production was observed after only 8 h of activation. Maximal levels of IL-2 and IL-4 were found 20 h after the onset of the stimulation which subsequently decreased. In contrast, IFN-gamma levels continued to increase in a period up to 40 h and then leveled off. In spite of these differences in secretion, the kinetics of...
Immunology
A mutant strain of rat, LEC, exhibits a novel maturational arrest of T cells in the thymus, that is, an arrest of the differentiation from CD4+ CD8+ to CD4+ CD8-cells. However, CD4+ T cells were found to arise in lymph nodes (LN) of LEC rats. These CD4+ T cells were T-cell receptor (TcR)/ CD3+ and the cross-linking of either TcR alone or both TcR and CD4 molecules induced [3H]thymidine deoxyribose (TdR) incorporation, similar to functional CD4+ T cells. Stimulation with a T-cell mitogen, concanavalin A (Con A), also induced the [3H]TdR incorporation and interleukin-2 receptor (IL-2R) expression in LEC rat CD4+ T cells. Nevertheless, Con A stimulation did not induce IL-2 production by these cells. However, when the stimulation signals were bypassed by ionomycin and phorbol 12-myristate 13-acetate (PMA), IL-2 was normally produced, suggesting that the IL-2 gene and nuclear factors for the IL-2 gene transcription are normal in LEC rats. These results suggest that signals induced by Con A are separated into the one for IL-2 secretion and the other for cell activation including DNA synthesis and IL-2R expression, and that the former signal transduction is blocked in LEC rat CD4+ T cells. Thus, CD4+ T cells in LEC rat LN might provide a good tool for investigating the regulatory mechanisms of IL-2 production.
Cellular Immunology, 1985
Concanavalin A (Con A), cloned interleukin 2 (&2), purified interleukin 1 (ILl) or two different crude preparations containing ILl activity alone, did not induce proliferation of rigorously accessory cell (AC)-depleted splenic L3T4+ or Lyt 2+ lymphocytes. Con A together with saturating concentrations of cloned IL2 (100 U/ml) promoted less than 40% of the proliferative responses observed in AC-supplemented L3T4+ and Lyt 2+ T-cell cultures. The three preparations of ILl used supported minimal proliferation of Con A-treated purified L3T4+ or Lyt 2+ lymphocytes. However, all these ILl preparations promoted significant growth of the T-cell populations if AC (1%) were included in the cultures. Cloned IL-2 combined with purified IL-1 promoted proliferation of Con A-treated L3T4+ and Lyt 2+ lymphocytes achieving-75% of the responses observed in AC-supplemented T-cell cultures. The additive effect of ILl was apparent in the presence of saturating concentrations of cloned IL-2. Finally, Con A alone induced a detectable number of both L3T4+ and Lyt 2+ lymphocytes to express IL2 receptors as determined with the anti-mouse IL-2 receptor antibody 7D4 by immunofluorescence and FACS analysis. Purified IL-1 neither induced detectable number of L3T4+ or Lyt 2+ T cells to express IL-2 receptors nor increased the number of Con A-treated T cells bearing IL-2 receptors. We have interpreted these findings to indicate the following: (a) Con A alone is sufficient to induce highly purified L3T4+ and Lyt 2+ lymphocytes to express IL-2 receptors. (b) Cloned IL-2 and purified IL1 are required for optimal growth of L3T4+ and Lyt 2+ lymphocytes and these cytokines together efficiently replace AC in growth of T cells initiated by Con A. IL-1 alone does not replace AC in Con A-induced activation of mouse T cells. (c) ILl exerts potentiation on IL2driven growth of Con A-treated L3T4+ and Lyt 2+ lymphocytes. The additive activity of IL1 on growth of normal T cells is not due to increased production of IG2 in the cultures or induction of normal T cells to expression of IL-2 receptors by IL-1. We propose that IL 1 optimizes the action and/or interaction of IL-2 with its receptors on the T-cell membrane (by, i.e., increasing affinity of the IL-2 receptor for its ligand and/or stabilizing the IL-2 raptor). 0 1985 AC&& PISS, 1~. ' The Base1 Institute For Immunology was founded and is supported by F. Hoffmann-LaRoche Ltd. Co.
Cellular Immunology, 1982
Interleukin 2 (IL 2) or T-cell growth factor induced the production of immune interferon (1FN-r) in C57B1/6 mouse spleen cell cultures, and enhanced mitogen-induced IFNy production in both spleen cells and thymocytes. Staphylococcal enterotoxin A, but not phytohemagglutinin P (PHA-P), induced 1FN-r production in thymocytes. IL 2 enhanced this production by almost 12-fold, while having no effect on the negative response to PHA-P. The IFN activity was shown to be IFNy by neutralization with specific antiserum. A strict correlation between IL 2 induction or enhancement of IFNy production and cell proliferation was not observed, probably indicating that non-IFN-producing cells also proliferated in the presence of IL 2. The data indicate that IL 2 can both induce IFNy and modulate mitogen induction of IFNr.
Journal of Experimental Medicine, 1994
Interleukin 12 (IL-12) facilitates the generation of a T helper type 1 (Th1) response, with high interferon gamma (IFN-gamma) production, while inhibiting the generation of IL-4-producing Th2 cells in polyclonal cultures of both human and murine T cells and in vivo in the mouse. In this study, we analyzed the effect of IL-12, present during cloning of human T cells, on the cytokine profile of the clones. The culture system used allows growth of clones from virtually every T cell, and thus excludes the possibility that selection of precommitted Th cell precursors plays a role in determining characteristics of the clones. IL-12 present during the cloning procedures endowed both CD4+ and CD8+ clones with the ability to produce IFN-gamma at levels severalfold higher than those observed in clones generated in the absence of IL-12. This priming was stable because the high levels of IFN-gamma production were maintained when the clones were cultured in the absence of IL-12 for 11 d. The CD4...
The Journal of Immunology
The cytolytic potential of a total number of 118 CD4+ human T cell clones specific for purified protein derivative (PPD) from Mycobacterium tuberculosis, tetanus toxoid, Lolium perenne group I allergen (Lol p I), Poa pratensis group IX allergen (Poa p IX), or Toxocara canis excretory/secretory antigen(s) (TES) was assessed by both a lectin (PHA)-dependent and a MHC-restricted lytic assay and compared with their profile of cytokine secretion. The majority of clones with Th1 or Th0 cytokine profile exhibited cytolytic activity in both assays, whereas Th2 clones usually did not. There was an association between the cytolytic potential of T cell clones and their ability to produce IFN-gamma, even though IFN-gamma produced by T cell clones was not responsible for their cytolytic activity. IL-4 added in bulk culture before cloning inhibited not only the differentiation of PPD-specific T cells into Th1-like cell lines and clones, but also the development of their cytolytic potential. The d...