Figure 1. PTPN22 regulates T cell dependent IL-17 responses induced by curdlan stimulated BMDC in vitro.

Wild type (WT) and Ptpn22^−/− derived bone marrow derived dendritic cells (BMDC) were pulsed overnight with OVA_323-339 (50nM) in the presence or absence of curdlan (100μg/ml) and co-cultured with OT-II T-cells. Cell-free supernatants were assessed for IL-17, IFNγ, and TNFα production by immunoassay on day 3 (A) and day 6 (B). Each point represents independent WT (closed triangle) or Ptpn22^−/− (open circle) BMDC preparations, connecting lines between WT and Ptpn22^−/− BMDC samples are paired by the same WT OT-II T-cell preparation (A) N=4, (B) N=7; NS = not significant, *p<0.05, **p<0.01, ***p<0.001 by two-way ANOVA, applying Sidak’s multiple comparisons test. (C-D) T-cells co-cultured for 6 days with WT or Ptpn22^−/− BMDC were harvested and replated in IL-2 and IL-23 for a further 4 days. At day 10 cells were restimulated for 6 hours with PMA and ionomycin in the presence of monensin and intracellular expression of IL-17 and IFNγ determined by flow cytometry. (C) One representative cytometric dot plot of 5 independent experiments. (D) Pooled data of N=5 independent experiments and represent mean + s.e.m.; NS = not significant, **p<0.01 by two-way ANOVA, applying Sidak’s multiple comparisons test.