FIG. 8.

The interferon gamma response to p199RY by PBL from Mamu-A*01⁺ Mamu-A*02⁺ SHIV-infected rhesus monkeys constitutes a significant portion of the response to all peptides spanning Env and Gag, as measured by Elispot analysis. PBL from 483 and 833 (two SHIV-infected Mamu-A*01⁺ Mamu-A*02⁺ rhesus monkeys) were added at 2 × 10⁵ per well to 96-well plates that had been coated with a mouse anti-human interferon gamma antibody. Peptide or medium alone was added in triplicate wells and incubated at 37°C for 18 h. Peptides used were p199RY or p199RD as a negative control at 8 μg/ml; two pools containing a combined total of 50 overlapping 20-mers at 8 μg/ml each, representing the entire SIVmac239 Gag protein; and two pools containing a combined total of 85 overlapping 20-mers at 8 μg/ml each, representing the entire HIV-1 89.6P Env protein. Plates were sequentially washed and incubated with biotinylated polyclonal anti-human interferon gamma, streptavidin-alkaline phosphatase, and chromogenic substrate, as described in Materials and Methods. The number of spot-forming cells per 10⁶ PBL is shown as the mean ± standard deviation of triplicate wells.