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. 2015 Oct 14;195(10):4822–4831. doi: 10.4049/jimmunol.1501828

FIGURE 5.

FIGURE 5.

PRN694 treatment decreases Treg frequencies and IL-2 production from CD4+ T cells in vitro and in vivo. (A and B) CD4+ T cells from spleen, mLNs, IELs, and LP from vehicle-treated or PRN694-treated mice at 7 wk posttransfer were examined for Foxp3 expression. (A) Dot plots show CD4 versus Foxp3 staining, with numbers indicating the percentages of CD4+Foxp3+ Tregs, and (B) the graph shows a compilation of data from three to five mice in each group. (C and D) Naive CD4+ T cells were stimulated in Th-polarizing conditions with PRN694 at the indicated concentrations for 72 h. Cells were then restimulated with PMA and ionomycin for 5 h and then analyzed for IL-2 production. (C) The percentages of IL-2–producing CD4+ T cells (gated on CD4+CD44hi) are shown. (D) Compilation of data from three independent experiments is shown, with the data for each Th subset normalized to the percentage of positive cells in the absence of inhibitor. (E and F) Isolated lymphocytes from vehicle-treated or PRN694-treated mice at 7 wk posttransfer were stimulated with PMA and ionomycin and analyzed for IL-2 production. Plots show gated CD4+CD44hi T cells, and the graph shows a compilation of data from three to five mice in each group. *p < 0.05, **p < 0.01.