Fig. 6: IL-1β is necessary for TH17 cell differentiation during antigen presentation.
From: T cell fate following Salmonella infection is determined by a STING-IRF1 signaling axis in mice
a BMDCs from wild-type and Irf1−/− mice were stimulated with OVA plus c-di-GMP for 18 h and washed three times with medium and co-cultured with MACS-sorted naïve OT-II cells in the presence of 5 ng/ml IL-1α or 5 ng/ml IL-1β. a IL-17A and IFN-γ production was measured in supernatants by ELISA at day 4. n = 4. **p < 0.01, ***p < 0.001. b, c BMDCs from wild-type and Irf1−/− mice were stimulated with OVA plus c-di-GMP in the presence of 20 ng/ml IL-23, 5 ng/ml IL-1β, and/or 10 μM PGE and washed three times with medium and co-cultured with MACS-sorted naïve OT-II cells in the presence b or absence c of IL-23, IL-1β, and/or PGE. IL-17A production in cell supernatants of BMDC:T cells co-cultured for 4 days was measured by ELISA. n = 3. *p < 0.05, **p < 0.001. d IL-23 and IL-1β treated CD4+ T cells from co-cultured Irf1−/− BMDCs were sorted at day 4 by flow cytometry. mRNA expression of the indicated genes was assessed by qRT-PCR. n = 3. *p < 0.05, **p < 0.01, ***p < 0.001. e wild-type and Irf1−/− mice were immunized with OVA plus c-di-GMP through i.p. route and recombinant mouse IL-1β were injected on day 1 and day 3 and sacrificed on day 5. CD4+ T cells were sorted from MLNs by MACS separation kit. qRT-PCR analysis of Il17a, RoRγc, and Ifnγ expressions in CD4+ T cells from Irf1−/− verses wild-type mice. n = 4. *p < 0.05, **p < 0.01, ***p < 0.001. p-values, one-way ANOVA d or two-way ANOVA a, b, e followed by Tukey’s multiple-comparisons test or unpaired two-tailed Student’s t-test a–c. Data shown represent two independent experiments.
