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. 2019 Dec 3;7(4):204. doi: 10.3390/vaccines7040204

Figure 3.

Figure 3

Polyfunctionality and phenotype of E1/E2-specific T cells as determined by intracellular cytokine staining of splenocytes of immunized mice. C57Bl/6 x Balb/c F1 mice were immunized i.m. with HCV E1/E2 protein (1 µg) with or without adjuvant on days 0, 21, and 35. Splenocytes were harvested on day 42 (n = 5/group) and analyzed by intracellular cytokine staining (ICCS) when stimulated by an E1/E2 peptide pool or media alone. The number of E1/E2-specific IFN-γ+ (A), TNF-α+ (B), and IL-2+ (C) per million CD4 T cells were determined. Values obtained with media alone were subtracted from those measured in the presence of the peptide pool. Correlation of IFN-γ responses as determined by ELISpot and ICCS was also performed (D). The frequency of cells expressing IFN-γ, TNF-α, and IL-2 alone or in combination is displayed (geomean per group) with the total number of cytokine-positive cells per million CD4+ T cells indicated below the pie chart (E). Meanings of the asterisks show in Section 2.8.