Fig. 4.

Suppressive activity and IL-17 secretion by CD25+ T cells based on GARP expression. (A) Experimental setup was as follows: CD4⁺ T cells were first sorted into CD25+ and CD25− subsets. The cells were then activated with anti-CD3/28 beads for 2 days, stained with GARP antibody, and sorted into GARP− and GARP+ cells. These cells were further expanded and rested in culture for an additional 10 days. (B) Suppressive activity of CD25+GARP+ and GARP− cells. Suppressive activity was performed and calculated as described in Fig. 1. Representative CFSE data are shown in Fig. S4. (C) Cells activated and sorted as shown in Fig. 4A were cultured in IL-2 for 8–10 days and then reactivated with PMA and ionomycin for 5 hours, followed by intracellular staining with IL-17–FITC, FOXP3.APC, and IFNγ-PE.Cy7 antibodies. (D) Expression of IFNγ versus IL-17 from the same staining shown in (C). Results are representative of three separate experiments from different donors.