Figure 8.

Expansion of Treg by IL-33 via modifying autophagy in vitro. Liver naïve CD4+ T cells (CD4⁺ CD25⁻ CD44⁻ CD62L⁺) were sorted from the infected WT mice at the 8th week post infection. T cells were cultured (3×10⁵ cells/well) in 96-well plates coated with anti-CD3 mAb (5 μg/mL, eBioscience) and soluble anti-CD28 mAb (3 μg/mL, eBioscience) and kept in presence of IL-2 (100 U/mL, eBioscience). Exogenous rmIL-33 (10 ng/mL) was added to investigate its ability to polarize Treg and PBS used as the control. Cells were cultured in RPMI-1640 Medium, 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL of Penicillin/Streptomycin for 5 days. The mRNA expression level of Treg-related parameters, Foxp3 (A) and TGF-β1 (B) and IL-10 (C) in cultured cells. The mRNA expression level of autophagy-related parameters, Atg5 (D) and Beclin-1 (E) and p62 (F) in cultured cells. Data are expressed as means ± SEMs based on 6 samples in each group and from 3 independent experiments. Asterisks mark significant differences among different groups (***P < 0.001).