Characterization of the immune response following AAV HIV Gag vaccination. Gag tetramer responses were monitored over time in CB6F1 mice that received i.m. administration of AAV8, AAVrh32.33, or SAdV24 encoding HIV Gag at two doses, 3 × 109 (A) and 3 × 1010 (B) GC. (C) Phenotypic characterization. Three weeks following AAV vector administration, lymphocytes were isolated from whole blood and spleens of mice injected with AAV2/AAV8, SAdV24, and AAV2/AAVrh32.33 Gag-expressing vectors (3 × 1010 GC). Per lymphocyte source, on the left, a scatter plot of CD8+ T cells of a representative mouse (n = 5) illustrates the tetramer positivity (Tet+), as quantified by the percentage of Tet+ within the CD8+ population. On the right, CD62L marker expression versus that of CD127 is represented. Memory phenotype is represented with the Tet+/CD8+ population overlaid in red on the total lymphocyte population in black. The percentages of Tet+/CD8+ T cells and TEM+/Tet+ T cells are indicated within the individual panels. (D) Gag-specific antibody responses. Sera from mice immunized with AAV8, AAVrh32.33, or SAdV24 at the dose of 3 × 1010 or 1 × 1011 GC were analyzed for anti-Gag IgG, and the results are presented as means with standard deviations (n = 4).