Figure 2.

Characterization of IL-27-stimulated polyclonal iTregs. Sorted naïve CD4 T cells (CD4⁺Foxp3^GFP−CD44^low) were cultured under Treg-skewing condition with immobilized anti-CD3 (2 μg/ml), anti-CD28 (2 μg/ml), IL-2 (100 U/ml), and TGFβ1 (5 ng/ml) for 3 days. Polyclonal iTregs were sorted based on GFP expression and restimulated with immobilized anti-CD3 (2 μg/ml), anti-CD28 (2 μg/ml), IL-2 (100 U/ml) in the presence or absence of rIL-27 (10 ng/ml) for 48 h. (A) Gene expression was measured by qRT-PCR. Data were normalized to untreated group. (B) Surface expression was measured by Flow cytometry. (C) Suppressive activity of sorted iTregs. Histogram represented one of three independent experiments. Data are mean ± SEM. *p < 0.05.