Figure 5: Adiponectin negatively regulates IL-17 production from dermal γδ-T cells through AdipoR1.

(a) Sorted dermal γδ-T cells were analysed for the expression of adiponectin receptors, AdipoR1 and AdipoR2. Mouse heart and liver extract were used as a positive control, respectively. (b,c) Dermal γδ-T cells were pretreated with indicated concentrations of adiponectin for 1 h, followed by treatment with IL-1β (10 ng ml⁻¹) and IL-23 (5 ng ml⁻¹) (b) for another 24 h, and the supernatant IL-17 levels were determined using the IL-17 ELISA kit (n=5: cells without IL-1β and IL-23 stimulation, n=6: cells with IL-1β and IL-23 stimulation), and (c) for another 6 h, and the gene expression levels of IL-17A was determined by quantitative (qPCR) (n=4: cells without IL-1β and IL-23 stimulation, n=5: cells with IL-1β and IL-23 stimulation). Data are mean±s.e. and are representative of two independent experiments. (d,e) Dermal γδ-T cells were transfected with AdipoR1 or AdipoR2 siRNA or nontargeting control siRNA. After 72 h, γδ-T cells were pretreated with adiponectin (10 μg ml⁻¹) for 1 h, followed by treatment with IL-1β (10 ng ml⁻¹) and IL-23 (5 ng ml⁻¹) for another 6 h. Messenger RNA expression of (d) AdipoR1 and AdipoR2 (n=8) and (e) IL-17A (n=4: cells without IL-1β and IL-23 stimulation, n=6: cells with IL-1β and IL-23 stimulation) in transfected cells were examined by qPCR. Data are mean±s.e. and are representative of three independent experiments. Data are analysed by Welch’s t-test for b,c,e. NS, not significant.