FIG. 1.

Magnitude of B*2705 epitope-specific CD8⁺ T-cell responses and restriction of a B*2705 Pol epitope. (A) OLP and B*2705 optimal peptide IFN-γ ELISPOT responses from subject R039. OLP 265 contains the B*2705 peptide binding motif: Arg at P2 and a hydrophobic or positively charged residue at the C terminus. (B) Peptide-pulsed BCLs, matched at each HLA allele to the HLA type of the donor subject, were incubated with a KY9-specific CD8⁺ T-cell line cultured from PBMCs from the donor subject. Only the HLA-matched BCLs and the autologous BCLs elicited a response measured by the production of MIP-1β by intracellular cytokine staining. (C) Truncations of the predicted epitope were titrated in serial log dilutions in an IFN-γ ELISPOT assay and run in triplicates. Error bars show standard errors of the means (SEM). The epitope with the lowest 50% effective concentration (EC₅₀) value is confirmed as the optimal epitope. (D) PBMCs from nine HIV-infected B*2705-positive subjects were screened by IFN-γ ELISPOT assay for the recognition of six known B*2705 epitopes (see Materials and Methods). Responses of >80 SFC per million cells are shown. (E) Median IFN-γ SFC per million PBMCs for KK10 (n = 9) and KY9 (n = 9) were compared. A Student's t test was performed (P = 0.438).