Fig 4.

Differential regulations of IL-12/IL-23 expression and IL-17 expression in CD14⁺ monocytes or PBMCs cocultured with HCV⁺ hepatocytes occurs through Tim-3 signaling. (A) Purified CD14⁺ monocytes were incubated with HCV^+/− Huh7 hepatocytes for 48 h, followed by flow cytometric analysis of Tim-3 expression on monocytes. Representative histogram and means + the SD from six repeated experiments for Tim-3 expression on monocytes incubated with HCV⁺ versus HCV⁻ hepatocytes are shown. *, P < 0.05 (as determined by Student t test). (B) Purified CD14⁺ monocytes were incubated with HCV^+/− Huh7 hepatocytes for 48 h, with αTim-3 or IgG antibody for another 72 h, and then stimulated with LPS/R848 for 6 h, followed by flow cytometric analysis for IL-12p35 and IL-23p19 expression. The data are shown as the results of a representative experiment (left) and means + the SD from six independent experiments (right). *, P < 0.05 (as determined by Student t test). (C) PBMCs were incubated with HCV^+/− Huh7 hepatocytes for 48 h, with αTim-3 or IgG antibody for 72 h, and then stimulated with PMA/ionomycin for 6 h, followed by flow cytometric analysis for IL-17A expression by CD4⁺ T cells. The data are shown as the results of a representative experiment (left) and means + the SD from six independent experiments (right). *, P < 0.05 (as determined by Student t test).