FIG. 7.

Ad5 retro-DAF–displaying vectors induce transgene (HIV-gag)–specific T-cell responses with a dramatically reduced magnitude and breadth. To test the magnitude and breadth of HIV-Gag–specific T-cell responses, splenocytes from Ad-treated (n = 5) and naive (n = 2) BALB/c WT mice were collected at 14 dpi (one dose; left) and 28 dpi (two doses; right), stimulated ex vivo (IFNγ ELISpot) with either Gag-specific peptide AMQMLKETI (A) or additional peptides and proteins, including irrelevant GFP protein as nonspecific control (B). Columns represent means ± SD. Statistical analysis was completed using two-way ANOVA with a Bonferroni post hoc test (stimulations × treatments); p < 0.05 was deemed a statistically significant difference. *p < 0.05, **p < 0.01, statistically different from naive mice; ^##p < 0.01, statistically different from WT Ad5-Gag treatment group within the same stimulation group. (C) To more fully test the breadth of HIV-Gag–specific T-cell responses, splenocytes collected at 28 dpi from Ad-treated (n = 5) and naive (n = 2) BALB/c WT mice were pooled and stimulated ex vivo with a pool of three 15mer peptides, spanning the complete HIV-Gag protein sequence (excluding peptides, used for individual stimulation). (Inset) Graph indicates the number of wells with >20 SFCs [arbitrary threshold (Appledorn et al., 2010)].