Figure 3
Fine-mapping of one novel epitope within Protease and two within Integrase. Peptide-specific CD8 cell lines were generated for three peptides, Protease 6, Integrase 17, and Integrase 29/30. PBMC collected from subjects with strong responses by ELISPOT were expanded using a bispecific CD3/4 antibody. Following expansion, peptide-specific cells were collected using an IFN-γ catching assay after stimulation with the appropriate peptide. Peptide specificity was confirmed by flow cytometry. HLA-restriction was then determined using peptide-pulsed target cells matched at only one MHC class I allele in a 51Cr-releasse assay at an E:T ratio of 10:1; peptide-pulsed autologous cells were used as a positive control. The sequences of the optimal epitopes were also determined by testing peptide-specific cell lines against serial dilutions of truncations of the original peptide in an ELISPOT assay. Data are shown for three epitopes: (A, B) – Protease 6. (C, D) – Integrase 17. (E, F) – Integrase 29/30.