Figure 3.

Analysis of the proliferation and intracellular cytokine production of IFNα subtype-stimulated Influenza-specific CD8⁺ T cells. Positively enriched Cell Trace^TM Violet-labeled CD8⁺ T cells from CL4 TCRtg mice were co-cultured with HA peptide-loaded BM-DCs in the presence or absence of IFNα4, IFNα6, or IFNα9 for 72 h (500 units/well). CD8⁺ T cell proliferation was measured as loss of cell tracer dye by flow cytometry. (A) Individual frequencies of proliferating CD8⁺ T cells and mean values (+SEM) are shown as dots and bars (n = 12). Multi-parametric flow cytometry was used to determine percentages and MFI of the intracellular expression of (B) IFNγ, (C) IL-2, (D) TNFα, and (E) GzmB in CD8⁺ T cells indicated as mean values (+SEM). Statistically significant differences between the IFNα subtype-stimulated group and the unstimulated group were analyzed by Ordinary One Way ANOVA analysis and Dunn's multiple comparison and are indicated by * for p < 0.05; ** for p < 0.01; *** for p < 0.001.