Egr3 TG γδ T cells produce IL-17 and skew CD4+ T cells to a Th17 phenotype.

<p>A) Splenocytes were stimulated and rested, and IL-17 production by CD8 γδ and DN γδ T cells was measured on day 6 after short restimulation (flow plots). The percentage of each subset producing IL-17 (top bar graph) and absolute number of IL-17 producing cells of each subset (bottom bar graph) are shown as an average of 4 independent experiments, each using 2 mice per genotype. B) Purified CD4+ T cells or γδ T cells were stimulated directly <i>ex vivo</i> with plate bound anti-CD3 and soluble anti-CD28 for 16 hours. IL-17A, IL-17F, IL-6, and IL-23R mRNA expression was measured by qRT-PCR done in triplicate and normalized to 18s expression. Data are representative of 3 independent experiments, each using 2 mice per genotype. C) IL-17 production measured as a percentage of either the Vγ1-expressing subset (left panels) or the Vγ4-expressing subset (right panels) of γδ T cells in splenocyte stimulation. Data are representative of 1 experiment using 5 mice per genotype. D) Egr3 WT CD4+ T cells were stimulated for 2 days with plate bound anti-CD3 and soluble anti-CD28 either alone (Th0), with the skewing cytokines TGFβ, IL-6, and IL-23 (Th17), or with equal numbers of Egr3 TG γδ T cells. IL-17 production by was assessed on day 6 after restimulation. CD4 and Thy1.1 expression shows the gating strategy used in this experiment (top row). IL-17 expression by the CD4+ Thy1.1- gated Egr3 WT T cells (bottom row). The overlay flow plot (top row, right) shows both the Egr3 WT CD4+ Thy1.1- T cells and Egr3 TG Thy1.1+ gates from the co-culture. Plots are representative of 3 independent experiments, which are averaged in the bar graph (bottom row, right). *p≤ 0.05, ** p≤ 0.01, ***p≤ 0.005