Figure 3. rHRV-DNA vaccination elicits superior poly-functional CMI in the gut mucosa.

Mice were vaccinated as described in the legend to Fig. 1 and lymphocytes prepared from mesenteric lymph nodes harvested 14 days after the final dose. The lymphocytes were stimulated for 1 h with 5 μg/ml of the H-2K^d-restricted Gag_197–205 immuno-dominant peptide and then cultured in the presence of Brefeldin A for a further 6 h. Cytokine production was analyzed by flow cytometry. We gated on CD8⁺ T cells to assess the number of CD8⁺ T cells producing (A) IFN-γ, (B) TNF-α, (C) IL-2, (D) IFN-γ and TNF-α, (E) TNF-α and IL-2, (F) IFN-γ and IL-2 and (G) IFN-γ, TNF-α and IL-2 after stimulation. The data are representative of 2 independent experiments, plotted as mean (n = 7) ± SEM and each symbol represents an individual mouse. *p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001 (Mann–Whitney U test).