Figure 5. Suppressive Foxp3⁺IL-17A⁺ cells are metabolically active.

(a,b) YFP⁺Foxp3^negIL-17A⁺, YFP⁺Foxp3⁺IL-17A⁺, YFP⁺Foxp3^negIL-17A^neg, YFP⁺Foxp3⁺IL-17A^neg and YFP^negFoxp3^negIL-17A^neg, YFP⁺Foxp3^negIL-17A^neg CD3⁺ T cells were sorted using the strategy presented in Fig. 4e and analysed with XFe Extracellular Flux Analyzer. (a) Cumulative data of two independent experiments evaluating basal respiration (OCR) versus glycolysis (basal extracellular acidification rate (ECAR)) of individual Th17–T_reg subsets (mean values of each real-time run, n=2 for YFP⁺IL-17A⁺Foxp3^neg, YFP⁺IL-17A⁺Foxp3⁺, YFP⁺IL-17A^negFoxp3^neg, YFP⁺IL-17A^negFoxp3⁺ and YFP^negIL-17A^negFoxp3⁺, n=7 YFP^negIL-17A^negFoxp3^neg). (b) CD4⁺ T cells, isolated from Cd45.1 mice and stained with CFSE, were analysed after 72 h of stimulation with αCD3 Ab, in the presence of irradiated CD4^neg fraction and Th17–T_reg subsets, sorted as presented in Fig. 4e. One-way analysis of variance (ANOVA) of immunosuppressive effects of the subsets at 1:4 ratio subset:CD4⁺. Similar results were obtained in an additional independent experiment. (c,d) The dose-dependent effects of 2DG (c) and etomoxir (d) on the transdifferentiation of Th17 cells (the percentage of IL-17A^negFoxp3⁺ cells of CD4⁺eYFP⁺) is shown. Similar results were obtained in an additional independent experiment. All data are mean±s.d. *P<0.05, **P<0.01 and ***P<0.001 by one-way ANOVA.