Figure 3

Wnt5a conditions DCs to drive regulatory T-cell cifferentiation. A. Schematic of in vitro Treg assay. B. Wnt5a-conditioned DCs stimulate CD4⁺FoxP3⁺ Treg differentiation in vitro following co-incubation with total splenic CD4⁺ T cells at a 1:1, 5:1 or 10:1 T cell to DC ratio. CD4⁺FoxP3⁺ Tregs were quantitated by flow cytometry. Representative of 3 independent experiments. C. DCs pre-conditioned for 24 and 48 hrs stimulate CD4⁺FoxP3⁺ Treg differentiation in vitro following co-incubation with naive splenic CD4⁺CD62L⁺ T cells. DCs were pretreated Wnt3a or Wnt5a for 24 or 28 hrs then co-incubated with splenic naïve CD4⁺ T cells with or without the IDO inhibitor, 1-MT. CD4⁺FoxP3⁺ Tregs were quantitated by flow cytometry. Performed in triplicate. Data is normalized to untreated DC condition. Representative of 3 independent experiments. P values based on a two-way ANOVA.