Functional characteristics of cytokine production capacities of HCV-NS3 antigen-specific CD8+ T-cells post boost with Ad5-, VV- or MVA-NS3/4A vectors.

<p>Freshly isolated splenocytes were stimulated with 0.1 ug/well of GAVQ peptide per 1.5 million cells and cultured in 10% FBS plus RPMI medium at 37°C for 5h. (A) Representative flow plots of intracellular staining of IFN-γ, TNF-α, and IL-2 in stimulated and unstimulated effector CD8+ T-cells (D7 post boost) and (B) memory precursor CD8+ T-cells (D28 post boost). Left panels show IFN-γ+/TNF-α+ co-producers and right panels show IFN-γ+/IL-2+ co-producers. Numbers in plots are representative percentages of cytokine producers. n = 3–4 mice per group. (C) Frequency (left) and mean fluorescence intensity (MFI, right) of IFN-γ producing CD8+ T-cells stimulated by GAVQpeptide. Error bars represent standard error of the mean. Asterisks represent significance analyses following post hoc Bonferroni testing between Ad5 and VV or Ad5 and MVA immunized mice. * = p value <0.05; ** = p value<0.01, *** = p value <0.0001.