IL-33 drives Tc9 cell differentiation in vitro. Naive CD8+ T cells were cocultured with BMDCs under Tc9-polarizing conditions with or without the addition of IL-33 for 2 days. Cell cultures without (Tc0) the addition of Tc9-polarizing cytokines TGF-β and IL-4 were used as controls. a, b Flow cytometry analysis of IL-9-expressing CD8+ T (Tc9) cells (a) and GzmB +CD8+ T cells (b). Numbers in the dot plots represent the percentages of CD8+IL-9+ T cells and GzmB +CD8+ T cells. Right, summarized results of three independent experiments obtained as reported on the left. c ELISA assessed IL-9 secretion in the cocultures. d–f qPCR analysis of the indicated cytokines (d), transcription factors (e) and St2 (f) in T cells. Expression was normalized to Gapdh and set at 1 in BMDC-induced Tc9 cells. g Naive CD8+ T cells from OT-I mice were cocultured with BMDCs under Tc9-polarizing conditions in the presence or absence of IL-33 for 2 days. B16-OVA-specific cytotoxicity of the cultured CD8+ T cells was examined. The results presented are the mean ± SD of 3–5 independent experiments. NS nonsignificant; *P < 0.05; **P < 0.01
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