FIGURE 3.

Cytokine profile of NS3-specific CD4⁺ T cells in vaccinated mice. Splenocytes from C57BL/6 mice, vaccinated with 10⁶ live or necrotic NS3 DC2.4 cells were harvested 14 days post-vaccination and restimulated with immunodominant NS3 epitopes. Intracellular cytokine staining was performed (Supplementary Figure 2) to assess the frequency of (A) CD3⁺ CD4⁺ T CD44^high effector memory T cells that are single cytokine producers of IFN-γ or IL-2 or TNF-α, (B) Frequency of CD4⁺ CD44^high effector memory T cells that are double cytokine producers of IFN-γ/TNF-α or IL-2/IFN-γ or IL-2-TNF-α. (C) The MFI of IFN-γ, IL-2 and TNF-α of NS3 specific CD3⁺ CD4⁺ T CD44^high T cells; (D) IFN-γ/TNF-α/IL-2 triple-positive CD4⁺ CD44^high effector memory T cells. Plots are representative of two independent experiments (n = 7/group). Data show mean ± SEM. ^∗p < 0.05, ^∗∗p < 0.01 (Mann–Whitney test).