IL-17 production in Th17 cell generated under the influence of PGE2 segregates with CCR6 expression. (A-C) CD4+ T cells were activated and expanded thereafter in the presence of irradiated allogeneic PBMCs and OKT-3 (0.1 μg/mL) in medium supplemented with IL-2, IL-23, and PGE2. CCR6+ (B) and CXCR3+ (C) were obtained from the parental Th17 cell line (A) by sorting using FACSAria. (D) Naive CD4+ T cells were activated by CD3 cross-linking in the presence of IL-12 and anti–IL-4 to obtain Th1 and expanded thereafter in the presence of IL-2. (A-D) The cells were harvested 10 days after sorting and processed for chemokine receptor expression and intracellular staining on PMA and ionomycin activation as described in “Flow cytometry.” The cells were all from the same patient and cultured for the same amount of time. The results here shown are representative of results generated with T-cell lines from two healthy persons.
