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. 2012 Dec 5;190(1):138–146. doi: 10.4049/jimmunol.1103721

FIGURE 3.

FIGURE 3.

CVT-6883 inhibits in vivo, but not in vitro Th17 differentiation. Splenocytes were isolated from EAE mice treated with CVT-6883 (3 mg/kg) or vehicle on day 10 PI and analyzed with flow cytometry. (A) Th1 and Th17 cells were analyzed by intracellular staining of IFN-γ and IL-17a, respectively, in the CD4+ gate. Data are mean ± SEM (n = 10), *p < 0.05 versus naive, #p < 0.05 versus vehicle. (B) qPCR analysis of Th1- and Th17-related gene expression in spleen. Data are mean ± SEM (n = 6), *p < 0.05 versus naive, #p < 0.05, ##p < 0.01 versus vehicle. (C–F) Splenocytes from naive and EAE mice treated with CVT-6883 (3 mg/kg) were restimulated in vitro with MOG35–55 for 48 h, and IL-17a (C), IFN-γ (D), IL-4 (E), and TGF-β (F) in supernatants were detected with ELISA. Data are mean ± SEM (n = 10), *p < 0.05 versus naive, #p < 0.05 versus vehicle. (G) Naive CD4+ T cells were induced to differentiate into Th1 or Th17 cells in vitro, in the presence of NECA (10 μM), CVT-6883 (100 nM), or the combination of the two. Data are mean ± SEM (n = 3), *p < 0.05 versus vehicle.