FIG 4.

Vaccination of rhesus macaques with viral vectors inclusive of a TAP inhibitor does not induce MHC-E-restricted T cells. (A) Macaques were vaccinated with an Ad5 prime followed by AAV1 boost at 12-weeks postpriming using one of the three inserts indicated: Gag-P2A-ICP47, Gag-P2A-UL49.5, or Gag-P2A-Rh185. An IFN-γ ELISpot assay was done weekly using peptide pools spanning the entire Gag protein. These responses were summed to give the total SIV Gag response. Additionally, minimal optimal peptides for SIV-specific responses restricted by Mamu-A*01 (CM9), Mamu-A*02 (GY9), and Mamu-E (RL9 and EK9) were included separately at each time point. (B) Representative ICS results of CD8⁺ T cells from the strain 68-1 RhCMV/Gag-vaccinated Mamu-A*01⁺ macaque Rh21826, the Ad5/Gag-P2A-UL49.5-vaccinated Mamu-A*01⁺ macaque Rh29804, and the Ad5/Gag-P2A-ICP47-vaccinated Mamu-A*01⁺ macaque Rh31520 are shown following coculture with the indicated antigen-presenting cells pulsed with the indicated peptide(s). E Min, a pool of 11 previously defined minimal optimal SIVmac239 Gag-derived peptides presented by Mamu-E (17). (C) Cumulative results from ICS assays using PBMC from two strain 68-1 RhCMV/Gag-vaccinated macaques and all six macaques that received an Ad5 prime/AAV1 boost regimen concomitantly expressing SIV Gag and a TAP inhibitor following incubation with the indicated antigen-presenting cells pulsed with the indicated peptide(s). The dashed line indicates the limit of detection (median background cytokine secretion) of the ICS assay. 11 Minimals, a pool of 11 previously defined minimal optimal SIVmac239 Gag-derived peptides presented by Mamu-E (17).