rAd induces polyfunctional memory CD4+ T cell responses. The DC subsets were exposed to different doses of rAd5-pp65 and rAd35-pp65 for 24 h before a 6-h coculture with autologous sorted T cells. A, Bars show the total frequencies of responding (IFN-γ, TNF-α, IL-2, MIP-1β, CD107a expressing) pp65-specific CD4+ T cells. Both DC subsets induced a greater magnitude of pp65 responses after exposure of rAd35-pp65 than to rAd5-pp65. B, The portions of the pies represent the 31 combinations of responses as described earlier and are shown grouped by number of functions: 5 (red); 4 (orange); 3 (yellow); 2 (green); and 1 (blue). The responses stimulated by rAd5 or rAd35 were compared with those found with overlapping (OL) peptide stimulation.