Figure 4.

SCFAs have opposite effects on IL-13-expressing T cells. Naïve CD4+ T cells (CD62L+CD44low) were sorted from spleens of FOXP3 GFP mice and cultured in 96-well flat bottom plates with plate bound anti-CD3 (2 μg/ml) and soluble anti-CD28 (0.5 μg/ml) in the presence of Th2 (IL-4 10 ng/ml, anti-IL-12 10 μg/ml, and anti-IFNγ 10 μg/ml) differentiation condition for 4 days. Butyrate and propionate (250 μM) were added in the culture at day 0. At day 4, cells were restimulated with with PMA (50 ng/ml), ionomycin (500 ng/ml), and brefeldin A (5 μg/ml) for 4 h at 37°C and 5% CO₂, and analyzed by flow cytometry (A). The frequencies of FOXP3+ and/or IL-13+ CD4+ T cells (B), IL-13+ and/or IL-5+ CD4+ T cells (C) and IL-13+ and/or IL-4+ CD4+ T cells (D) are represented by bar graphs. Data are shown as mean ± S.D. Two-way ANOVA and Tukey's multiple comparison test were used for statistical analysis. NS, not significant.