HAdV-5-specific CD8+T cell responses. CD8+ T cells were cultured as described in Figure 1, and were tested for responses to HAdV-5 and melanoma antigens. (A) The mean frequency of CD8+ T cells from 7 donors responding to HAdV-5 by ELISPOT is shown. (B) These cells were also tested for IFNγ and IL-2 production by flow cytometry. Data from 7 independent HD is shown. Identically cultured melanoma patient cells responding to HAdV-5 LacZ or HAdV-5 TMM2 (HAdV-5 plus melanoma antigens) is shown (n = 2, averaged), for total IFNγ-producing cells and polyfunctional, simultaneous IFNγ + IL-2-producing cells (C). Responses were highest at week 3 for 5 of 7 donors and 1 of 2 patients. *Significant response vs. baseline by single tailed student’s t-test (p < 0.05).