Fig. 2: Tolerogenic DCs generated via CB1- and PPARα-mediated activation induce functional FOXP3⁺ Tregs.

a Scheme of coculture experiments. b Cytokines produced by allogeneic naïve CD4⁺ T cells primed by unstimulated, WIN55212-2 (WIN, 10 µM), LPS (0.1 µg/mL) or LPS plus WIN55212-2-treated hmoDCs after 5 days (n = 8). c Cytokine ratios by primed naïve CD4⁺ T cells in the indicated conditions (n = 8). d Percentage of induced FOXP3⁺ Tregs under the indicated conditions (n = 8). Flow cytometry representative dot plots are shown. e Suppression effects of purified induced FOXP3⁺ Tregs generated by LPS/WIN55212-2-stimulated hmoDCs. f Percentage of induced FOXP3⁺ Tregs by allogeneic LPS or LPS/WIN55212-2-stimulated total DCs (n = 6). Flow cytometry representative dot plots are shown. g Geometric mean fluorescence intensity (gMFI) of CD83 expression and TNFα and IL-6 production after stimulation of hmoDCs with LPS (0.1 µg/mL) plus WIN55212-2 (10 µM) in the presence of CB1 (Rimonabant (RIM), 20 µM), CB2 (AM630, 20 µM), PPARα (GW6471, 25 µM) and PPARγ (GW9662, 10 µM) antagonists for 18 h (n = 8). h Percentage of FOXP3⁺ Tregs generated after 5 days by hmoDCs stimulated with the indicated conditions. Values are mean ± SEM. Statistical significance was determined using One-way Anova (b, d, g, h) or Paired t test (c, f). *P < 0.05, **P < 0.01, ***P < 0.001.