Simultaneous production of IL-2, IL-4, and IFN-gamma by activated human CD4+ and CD8+ T cell clones (original) (raw)
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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.
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.
Generation of rat TH2-like cells in vitro is interleukin-4-dependent and inhibited by interferon-γ
Immunology
Differentiation of naive T cells into effector cells producing T helper type 1 (Th1) and Th2 cytokines is regulated by the presence of specific cytokines in the T-cell microenvironment. The effect of interferon-γ (IFN-γ) and interleukin-4 (IL-4) on Th1- and Th2-like cell development was investigated in cultures of mixed rat spleen cells. These cells were cultured for 4 days in medium containing concanavalin A (Con A) with or without additional IL-2, IFN-γ or IL-4. The cells were then washed and their capacity to produce IL-4, IL-5 and IFN-γ determined following stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin. Freshly isolated cells stimulated with PMA and ionomycin expressed detectable levels of IL-4 and IL-5 mRNA as measured by a quantitative polymerase chain reaction (PCR) procedure and much higher levels of IFN-γ mRNA. Cells cultured with Con A for 4 days, washed, and restimulated with PMA + ionomycin were unable to express detectable levels of IL-4 and IL-5 ...
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...
Journal of Experimental Medicine, 1995
Naive T cells in the periphery mainly secrete interleukin (IL) 2 upon activation. After stimulation in the presence of appropriate costimulators, both CD4 + and CD8 + T cells differentiate into effector cells secreting distinct T helper (Th) l-and Th2-1ike cytokine patterns. Subsequent to differentiation, both CD4 + (Thl and Th2) and CD8 + (TC1 and TC2) cells are stable and cannot be induced to differentiate into the opposite pattern or revert to the naive cytokine secretion pattern. We now show that IL-4 caused committed TC1 bulk populations or clones to lose the ability to synthesize IL-2. The cells retained the ability to secrete interferon (IFN) % granulocyte/macrophage colony-stimulating factor, and tumor necrosis factor, did not synthesize any Th2 cytokines, and did not alter cell surface marker expression. IL-4 rapidly inhibited IL-2--synthesizing ability in the absence or presence of antigen-presenting cells, thus demonstrating that IL-4 acted direcdy on TC1 cells. The defect in IL-2 synthesis could not be reversed by subsequent stimulation with potent antigen-presenting cells in the presence of IL-2 and anti-IL-4, or with anti-CD3 plus anti-CD28 antibodies. Both IL-2 + and IL-2-TC1 cells were strongly cytotoxic toward allogeneic but not syngeneic targets. However, IL-2-TC1 cells were unable to proliferate unless exogenous IL-2 was provided. TC1 cells that lose IL-2 synthesis but retain IFN-'y synthesis and cytotoxicity may be similar to the "anergic" cells induced by stimulation of CD4 + or CD8 § cells in the absence of costimulators. These results suggest that during a mixed type 1/type 2 response in vivo, IL-4 may induce the IL-2 § TC1 -~ IL-2-TC1 conversion, and thus curtail the expansion of the TC1 response without impairing short-term effector function.
European Journal of Immunology, 1989
Tumor necrosis factor-alpha (TNF-a) is a pleiotropic lymphokine which may have important regulatory effects on immune responses. It is shown here that eight alloreactive CD4' T cell clones (TCC) secreted significant amounts of TNF-a after stimulation with either specific alloantigen or 12-0-tetradecanoylphorbol 13-acetate together with the calcium ionophore ionomycin (up to 50 ng/ml/24 W106 cells) whereas CD8' TCC failed to do so (max. 2 nglmV24 W106 cells). The CD8' TCC also secreted markedly less granulocyte/macrophage colony-stimulating factor than the CD4' cells. However, this was not indicative of a general decrease of lymphokine production by CD8' cells because CD4' and CD8+ TCC both secreted similar amounts of interferon-gamma. These results show that regulatory CD4' lymphocytes can produce large amounts of TNF-a, whereas CD8' effector cells cannot do so.
Induction of type 2 activity in adult human CD8+ T cells by repeated stimulation and IL-4
International Immunology, 2001
⍣ and CD30 ⍣ CD8 ⍣ T cells, up-regulated the number of IL-5 ⍣ cells, and increased IL-5 levels released. These observations demonstrate that repeated TCR-CD3 stimulation of normal human CD8 ⍣ T cells favoured the growth of cells with a type 2 phenotype and that this was further amplified by the presence of IL-4. These mechanisms may be important in virus-induced lung eosinophilic inflammation in healthy subjects and virus-induced exacerbations of asthma.
Proceedings of the National Academy of Sciences, 1988
We have investigated the linkage between CD4/CD8 phenotype and programing for specific responses in primary T-cell populations. In situ hybridization has been used to determine the frequency of cells competent to express the interleukin 2 (IL-2) gene after short-term stimulation with various polyclonal activators. The effects of the T-cell receptor ligands Con A and anti-CD3 monoclonal antibody were compared with those of a calcium ionophore that bypasses membrane receptors altogether. Induction with a calcium ionophore and phorbol ester revealed that potential IL-2 producers not only constitute >85% of the cells with a CD4' "helper/ inducer" phenotype but also constitute over half of the cells with a CD8' "killer/suppressor" phenotype. There is no defect in the ability of these CD8' cells to accumulate IL-2 transcripts under these conditions. By contrast, in response to phorbol ester and either Con A or anti-CD3, the CD8' cells show an abortive IL-2 production response with rapid disappearance of IL-2 mRNA. This results in substantially lower yields of IL-2 per cell than is made by CD4+ cells in response to the same stimuli. The extent to which these populations appear to have diverged in function thus depends on the stimulus used to trigger the response. The results suggest that differences in signal transduction or posttranscriptional regulatory mechanisms, rather than effector gene inducibility per se, may initially underlie the commitment of CD4+ and CD8+ cells to distinct functional roles. Mature T cells are functionally specialized in their responses to recognition of antigen. In general, T-cell lines that express the CD4 cell-surface glycoprotein are "helper" or "amplifier" cells that respond to the recognition of antigen by secreting lymphokines, often including the major T-cell growth factor interleukin 2 (IL-2) (1, 2). Cells that express CD8 include most killer T cells and show little helper activity (3-6). This suggests that the constitutive CD4/CD8 phenotypes of T cells are correlated with the inducibility of different, limited sets of functional response genes. Although the correlation is frequently observed in memory T cells and T-cell lines, it is not known how it is established during T-cell differentiation. When fresh CD8+ cells are activated and Abbreviations: IL-2, interleukin 2; mAb, monoclonal antibody; PMA, phorbol 12-myristate 13-acetate; nt, nucleotide.
Scandinavian Journal of Immunology, 1988
Ihe activation signals necessiiry for interleukin-2 (IL-2) reeeptor induction, lL-2 production, and DNA synthesis in resting T cells were investigated. IL-2 reeeptors were Indueed after aetivation via CD2 or CD3 alone, while IL-2 production in both CD4' and CD8' T cells required aetivation via both CD3 and CD2. The sequence of aeiivation signals via CD3 and CD2 was shown to be important since DNA synthesis was induced when the primary activation signal was delivered via CD3. and the CD2 signal within 8 h. In contrast, no DNA synthesis was demonstrated when the primary activation signal was delivered via CD2 and the CD3 signal later. Ciclosporin A (CyA) inhibited T-cell DNA syntljesis after activation via CD2 and CD3. The inhibition seemed to be due to the prevention of [L-2 synthesis.