Figure 3.

3×GM-CSF+rHBVvac immunization induced a strong Tc1 immune response and enhanced HBs-specific CTLs. (a) Th1, Tc1, Th17, Tc17, and Th2 cell induction in response to vaccination. Splenic T cells were isolated on day 14 after the third immunization, stimulated with rHBsAg (10 μg mL⁻¹) for 12 h in culture and stained intracellularly for IFN-γ, IL-4, and IL-17a in CD4⁺ or CD8⁺ T cells. (b) IFN-γ⁺ intracellular staining of CD8⁺ eFluor 670-labeled T cells after culture with rHBsAg (10 μg mL⁻¹) for three days. (c) The percentage of IFN-γ⁺ CD8⁺ eFluor 670^low T cells is summarized (d). In vivo cytotoxic lysis assays were performed on the 14th day after the third immunization. (e) The percentage of specific lysis is summarized. The data are shown as mean ± SEM (n = 5) from three independent experiments. *p < 0.05 and **p < 0.01 (unpaired Student's t-test).