Fig. 5. Regulation of IL-17 expression by TCR triggering and inflammatory cytokines.

(A) BioMark analysis of PE⁺ γδ T cells from the dLN of mice immunized with PE-alum or OVA-alum 60 hours prior, directly ex vivo (2 cells/sample); (B) IL-1 receptor expression on PE⁺ and PE⁻ γδ T cells from dLN of mice immunized with PE-alum 60 hour prior; (C) CD44, CD62L expression of the total spleen γδ T cells from the naive C57BL/6 mouse, and the expression of IL-1R and CD27 expression of each subpopulation according to their CD44, CD62L expression. (D) BioMark analysis of splenic CD62L^lo and CD62L^hi γδ T cells after in vitro stimulation with plate bound anti-CD3 (10 μg/ml), IL-1 and IL-23 (1ng/ml each), Pam₃Csk₄ (1μg/ml) and IL-23 (1ng/ml), or media alone. Mean and standard deviation of qPCR analysis of (E) IL-17 mRNA of total splenic CD62L^lo γδ T cells (purified by FACS sorting, achieving >98% purity), stimulated with plate bound anti-CD3 and anti-CD3 together with IL-1 and IL-23 (1ng/ml each), in the absence or presence of 100 ng/ml Cyclosporine A and (F) IL-17 mRNA of total γδ T cells enriched from G8 TCR transgenic mice lymph nodes by negative depletion (achieving >98% purity as determined by FACS) stimulated with IL-1 and IL-23 (1ng/ml each), plate bound anti-CD3 and anti-CD3 together with IL-1 and IL-23, at indicated time points. All results are representative of at least 3 independent experiments.