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. Author manuscript; available in PMC: 2018 Aug 1.
Published in final edited form as: J Immunol. 2017 Jun 26;199(3):941–954. doi: 10.4049/jimmunol.1601890

Figure 7. TLR7-mediated inhibitory effect of Th17 differentiation does not require DC involvement.

Figure 7

(A) Imiq and Lox significantly inhibited Th17 cell generation in the DC co-culture system. In contrast, LPS and Pam3 promoted the generation of Th17 cells in this system. Mouse naïve CD4+ T cells were co-cultured with DCs at a ratio of 10:1 with 1μg/ml soluble anti-CD3 in the absence or presence of indicated TLR ligands. IL-17- and IFN-γ-producing CD4+ T cells were evaluated using flow cytometry after stimulation with PMA and ionomycin for 5 hours. Results shown in the right panel are the summary (mean ± SD) of IL-17-producing cells in the DC co-culture system in the presence of indicated TLR ligands from 3 repeated experiments with similar results. *p<0.05 and **p<0.01, compared with the medium only group determined by paired t-test. (B) Separation of DCs from CD4+ T cells by a transwell insert dramatically decreased Th17 polarization in the Transwell system, but did not affect the suppression effect of Th17 cell differentiation mediated by Imiq. Naïve CD4+ T cells were separated from DCs using transwell inserts, and cultured in the Th17 differentiation medium in the absence or presence of Imiq. Results shown in the right panel are mean ± SD of IL-17-producing cells in the Transwell system from 3 repeated experiments with similar results. **p<0.01, compared with the medium only group determined by paired t-test. (C) and (D) Pre-treatment of DCs with TLR7 ligands did not influence Th17 differentiation either in coculture (in C) or in transwell systems (in D). Mouse DCs were pretreated with or without Imiq, Lox, Pam3 and LPS for 1 day, and then cocultured with CD4+ T cells (in C), or separated with naïve CD4+ T cells by a transwell insert (in D), in the presence of soluble anti-CD3 (1μg/ml) and Th17 polarization cytokines for 6 days. IL-17- and IFN-γ-producing CD4+ T cells were evaluated using flow cytometry after stimulation with PMA and ionomycin for 5 hours. Data shown in (A) to (D) are a representative from at least three individual experiments.