(A) Th17-polarized T cells were generated from naive Il17fThy1.1/Thy1.1 CD4+ T cells with Il12b−/− APCs and anti-CD3 (5 μg/ml) as in Figure 1, in the absence (Th17 -IL-23) or presence (Th17 +IL-23) of 5 ng/ml IL-23. Thy1.1+ (IL-17F+) cells were isolated by FACS sorting and 3 × 105 cells were transferred into congenic Rag1−/− mice as indicated. Recipients were administered 100 μg of anti-IL-23R, anti-IL-12p40, or isotype control via i.p. injection in saline 1 day prior and 1 day after T cell transfers and once weekly thereafter (five total doses). Mice were sacrificed 4 weeks after transfer and analyzed histologically and scored for development of colitis.
(B) Mice that received Th17 -IL-23 (IL-17F+) T cells and treated as in (A) were sacrificed 4 weeks after transfer, and cells recovered from MLNs were stimulated ex vivo with PMA ionomycin and analyzed for CD4 and intracellular IL-17A and IFN-γ by FACS. Data are the means from five mice in each treatment group. *p < 0.05 and **p < 0.01 versus control treatment group. Data are representative of two similar experiments.