PMC:2791889 / 8907-12038 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/2791889","sourcedb":"PMC","sourceid":"2791889","source_url":"http://www.ncbi.nlm.nih.gov/pmc/2791889","text":"IL-10 Production by Th1 Cells Is Dependent on STAT4 but Not on STAT6, IFN-γ, or IL-4 Signaling\nTo further elucidate the mechanisms required for the development of Th1 cells producing IL-10, we investigated the role of STAT4, one of the signaling pathways activated by IL-12 (Murphy et al., 2000). Naive CD4+ D011.10 T cells deficient in STAT4 (Ouyang et al., 1998) were cultured in the presence of IL-12 and OVA. Again, IL-10-producing Th1 cells were differentiated at the high antigen dose in the presence of IL-12 in DO11.10 T cells (Figure 2A). In contrast, in the absence of STAT4, the percentage of cells expressing IFN-γ was dramatically diminished as expected and resulted in an increase in the percentage of cells expressing IL-4, but not IL-10 (Figure 2A), suggesting that STAT4 contributes to IL-10 expression by Th1 cells.\nBecause IL-10 expression is associated with an IL-4-induced Th2 cell phenotype, we investigated whether the differentiation of the IL-10-producing Th1 cells depended on signaling through the IL-4 receptor via STAT6 activation (Glimcher and Murphy, 2000; Murphy et al., 2000). The absence of STAT6 did not impair the differentiation of IL-10-producing Th1 cells in the presence of IL-12 and OVA (Figure 2A). In fact, a higher percentage of STAT6-deficient cells compared with WT cells produced both IL-10 and IFN-γ (Figure 2A), which may be the result of the loss of Th2 cell control over a Th1 cell response. As expected, lack of STAT6 abrogated both IL-4 and IL-10 production by T cells developed with IL-4 or with low antigen dose (Figures 2B and 2C). However, in the absence of STAT4 signaling, IL-10 and IL-4 production by Th2 cells was if anything increased (Figures 2B and 2C). Thus, in contrast to what was observed under Th1 conditions, IL-10 expression by Th2 cells depended on STAT6, but not on STAT4, signaling (Figures 2B and 2C).\nTo investigate whether the inability of STAT4-deficient T cells to produce IL-10 might be due to the absence of IFN-γ, as suggested before (Shaw et al., 2006), we differentiated DO11.10 or DO11.10 IFN-γ-deficient naive CD4+ T cells in the presence of IL-12 and increasing doses of OVA. The secretion of IL-10 as induced by high antigen dose, and IL-12 was not affected by an absence of IFN-γ (Figure 2D), showing that the expression of IL-10 by Th1 cells is independent of IFN-γ. In the absence of IFN-γ, we observed an increase in the secreted IL-4 as expected (data not shown).\nWe also tested for any potential role of IL-4 in the development of Th1 cells producing IL-10 by culturing DO11.10 or DO11.10 IL-4-deficient naive CD4+ T cells with IL-12 and increasing doses of antigen. As observed in the absence of STAT6 (Figure 2A), IL-4 deficiency had no effect on the development of Th1 cells producing IL-10 (Figure 2E), but compromised the development of Th2 cells producing IL-10 (Figure 2F). Thus, our data suggested that IL-10 production by Th1 or Th2 cells was dependent on the specific signaling pathways required for their differentiation, given that STAT4 is required for the induction of IL-10 production by Th1 cells and STAT6 for Th2 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