FIGURE 1.

T_H17 cells are not sensitive to DEX treatment in vitro. CD4⁺CD62L⁺CD25⁻ naive T cells isolated from DO11.10 OVA TCR-transgenic mice were cultured with WT BALB/c splenocytes that had been pulsed with OVA_323–339 under polarizing conditions. On day 6, cells were collected and stimulated with PMA/ionomycin for precursor frequency by ELISPOT or pretreated for 2 h with the indicated doses of DEX before stimulation with CD3/CD28 beads + IL-2. A, Spot frequencies of IL-17-producing T cells. B, Spot frequencies of IL-4-producing T cells. Cells were plated under the following conditions: T cells indicate un-stimulated T cells cultured without bead stimulation; 1 μM DEX indicates T cells pretreated with 1 μM DEX but not stimulated with CD3/CD28 microbeads; beads indicates untreated T cells stimulated with CD3/CD28 microbeads; μM DEX + bead indicates T cells pretreated with the indicated dose of DEX and stimulated with CD3/CD28 microbeads. Cell culture supernatant was collected at 36 h after stimulation and the levels of T_H2 and T_H17 cytokines were determined by multiplex suspension array assay. C, IL-5 levels from T_H2 cells. D, IL-13 levels from T_H2 cells. E, IL-17 levels from T_H17 cells. F, IL-22 levels from T_H17 cells. All data are graphed as mean ± SEM for n = 5–8;*, p < 0.01.