(A–C) Higher ex vivo expression of TIGIT and different co-inhibitory molecules on virus-specific CD4+ T cells of acutely and chronically infected HCV patients compared to patients with spontaneously resolved HCV. (A) Representative dot plots depicting the ex vivo co-inhibitory receptor expression (TIGIT, PD-1, BTLA, TIM-3) of HCV-specific MHC class II tetramer+ CD4+ T cells of patients with acute, chronic and resolved HCV infection. (B) Frequencies of the inhibitory receptors TIGIT, PD-1, BTLA, and Tim-3 on total and HCV-specific CD4+ T cells of patients with acute, chronic, and spontaneously resolved HCV infection. P-values were calculated by the Tukey’s multiple comparison test. P-values smaller than 0.05 were considered significant, where *,** and *** indicate p-values between 0.01 to 0.05, 0.001 to 0.01 and 0.0001 to 0.001 respectively. (C) SPICE analysis of TIGIT, PD-1, BTLA, and Tim-3 co-expression pattern of HCV-specific CD4+ T cells of patients with acute (n = 10), chronic (n = 10), and spontaneously resolved (n = 8) HCV infection.