Fig. 2

Interleukin (IL)-35 stimulation to HBV core-specific CD4⁺CD25⁺CD127^dim/− regulatory T cells (Tregs) and T helper 17 (Th17) cells in chronic hepatitis B (CHB) and asymptomatic HBV carriers (ASC). CD4⁺ T cells were purified from eighteen CHB and sixteen ASC, and were stimulated with or without IL-35 for 12 h in the presence of HBc 18–27 peptide. IL-12p35 and EBI3 mRNA expressions were investigated in unstimulated CD4⁺ T cells by real-time PCR. a IL-12p35 mRNA in CD4⁺ T cells was comparable between CHB and ASC. b EBI3 mRNA in CD4⁺ T cells was remarkably elevated in CD4⁺ T cells from CHB when compared with ASC. Individual level for each subject was presented, and comparisons were made using Student t tests. HBV core-specific CD4⁺CD25⁺CD127^dim/− Tregs and CD4⁺IL-17⁺ Th17 cells with or without IL-35 stimulation were investigated by flow cytometry. c HBV core-specific CD4⁺CD25⁺CD127^dim/− Tregs percentage was increased in response to IL-35 stimulation in both CHB and ASC. d HBV core-specific CD4⁺IL-17⁺ Th17 cells percentage was comparable between IL-35 stimulated and unstimulated CD4⁺ T cells in both CHB and ASC. e Ratio of Tregs/Th17 was increased in response to IL-35 stimulation in both CHB and ASC. Individual level for each subject was presented, and comparisons were made using paired t tests. mRNA relative expressions corresponding to Tregs transcriptional factor FoxP3 and Th17 transcriptional factor RORγt were measured by real-time PCR. f FoxP3 mRNA was notably increased in response to IL-35 stimulation in both CHB and ASC. g RORγt mRNA did not change significantly in response to IL-35 stimulation in either CHB or ASC. Individual level for each subject was presented, and comparisons were made using paired t tests. IL-10 and IL-17 production in cultured supernatants were measured by ELISA. h IL-10 expression in cultured supernatants was notably elevated in response to IL-35 stimulation in both CHB and ASC. i IL-17 expression in cultured supernatants was remarkably decreased in response to IL-35 stimulation only in CHB, but not in ASC. Individual level for each subject was presented, and comparisons were made using Wilcoxon matched pairs tests. j phosphorylated STAT3 (pSTAT3), STAT3, and GAPDH expression in CD4⁺ T cells with or without IL-35 stimulation was investigated by Western blot. pSTAT3 was significantly down-regulated in CD4⁺ T cells in response to IL-35 stimulation, however, total STAT3 expression was comparable in CD4⁺ T cells with or without IL-35 stimulation