Figure 5. Renal impairment and Angiotensin II enhance TH17 polarization.

(A) mRNA expression of markers of T cell lineages (T-bet for the T_H1, GATA-3 for T_H2, RORγt for T_H17 and Foxp3 for T_reg) was assessed in spleens and aortas of Apoe^−/− mice after 12 weeks high fat diet (renal impairment relative to controls, n=5, t-tests). (D-E) Interferon gamma (IFNγ)(B,C, n=6-8) and IL-17A producing (B,D, n=8-13) T cells and Foxp3⁺ regulatory T cells (E, n=4-5) in spleens from Apoe^−/− mice with normal and impaired renal function (RI) (5h PMA/ionomycin, % of CD3⁺ cells, Il17a^−/− spleens used to define IL-17A positivity (<0.1%)). (F,G) Mouse splenocytes were cultured under T_H0, T_H1 and T_H17 polarizing conditions with and without exogenous Angiotensin II (AngII, 250 pg/ml unless indicated) and intracellular cytokine expression was assessed after re-stimulation as described in methods (F, typical examples and G, n=3 independent experiments, One-way ANOVA, *indicates significant slope). (H) T_H17 polarization was carried out in the presence or absence of Angiotensin II and Losartan (1nM) or solvent control (one of 2 independent experiments).