Figure 5
From: TH17 cells express ST2 and are controlled by the alarmin IL-33 in the small intestine
Mouse and human TH17 cells respond to IL-33 in vitro. (a) Mouse naive sorted CD4+CD25−CD62L+CD44− T cells were stimulated in vitro with anti-CD3/CD28 and grown under TH17 or Treg conditions for 5 days. ST2 expression was analyzed by qPCR and compared with naive cells (mean±s.d. of three independent experiments). (b) Mouse TH17 cells were treated with 20 ng ml−1 IL-33 during the differentiation process (as in a). Expression levels of Il10, Tbx21, Ifng, and Csf2 were quantified by qPCR (dotted line at 1 indicates the value of control untreated TH17 cells). Data are presented as mean±s.e.m. of ≥3 independent experiments. (c) Fresh ex vivo sorted human CCR6+CCR4+CXCR3–CD45RA–CD25–CD8– TH17 cells were stimulated with CD3 and CD28 mAbs for 48 h and expanded for another 3 days in the presence of 20 ng ml−1 IL-33. IL10 and TBX21 expression levels were quantified by qPCR (dotted line at 1 indicates value of non-treated TH17 cells). Human TH17 cells were also stimulated with PMA/Ionomycin for 5 h to detect intracellular levels of IL-17A, IL-10 and GM-CSF by FACS. One representative dot plot out of three independent experiments of flow cytometry analysis of IL-10 and GM-CSF assessed on TH17 cells (gated on IL17A+ CD4+ T cells) is shown in d. (e) Mouse TH17 cells were treated with 20 ng ml−1 IL-33 during the differentiation process (as in a) and the expression levels of Il10, Maf, Ahr, Prdm1, Irf4, Jun, and Ikfz3 were quantified by qPCR (dotted line at 1 indicates the value of control untreated TH17 cells). Data are presented as mean±s.e.m. of five independent experiments.
