FIG. 4.

SIV Nef peptide-specific cytolytic activity of PBL from infected Mamu-A*02⁺ rhesus monkeys is localized to the 9-amino-acid peptide p199RY. (A) PBL from 18324, 19242 (two SHIV-infected Mamu-A*01⁻ Mamu-A*02⁺ rhesus monkeys) and 19196 (an SIVmac251-infected Mamu-A*01⁺ Mamu-A*02⁺ rhesus monkey) were stimulated in vitro with p 199 (20 μg/ml). On day 12 of culture, the lymphocytes were assessed as effector cells in a standard 4-h ⁵¹Cr release CTL assay. Target cells were B-LCL made from monkey 19196 (used in experiments with effectors from monkey 19242 or monkey 19196) or monkey 24195 (Mamu-A*01⁻ Mamu-A*02⁺; used in experiments with effectors from 18324) that had been incubated overnight with p199 (20 μg/ml) or either of the 13-amino-acid peptides p199R or p199S. (B) PBL from monkeys 26795, 14282, and 19242 (three SHIV-infected Mamu-A*01⁻ Mamu-A*02⁺ rhesus monkeys) were stimulated in vitro with 5 μg of the SIV Nef 13-amino-acid peptide p199R per ml. On days 11 and 12 of culture, the lymphocytes were assessed as effector cells in a standard 4-h ⁵¹Cr release CTL assay. Target cells were B-LCL made from monkey 24195 that had been incubated overnight with 5 μg/ml of either SIV Nef p199R or one of the SIV Nef 9-amino-acid peptides p199RD, p199RY, or p199RT.