Identification of novel epitopes using peptide pools. (A) Workflow used to identify novel immunogenic peptide candidates. (B) Representative flow cytometric density plot analyzing IFN-γ⁺/CD8⁺ T-cell frequencies after stimulation with canonical peptide pools D and N as well as a negative background control (HIV-1 p17 HLA-B∗15). (C) Overview of peptide pools A-N (consisting of 7 peptides each). Each single peptide was present in 2 pools (A-G and H-N). Peptide sequences are highlighted if both peptide pools containing a peptide elicited IFN-γ⁺/CD8⁺ T-cell frequencies of ≥0.02%. Color density (0%-30% donor reactivity) indicates the percentage of healthy donors with a cytotoxic T-cell response induced by a certain peptide pool (IFN-γ⁺/CD8⁺ T-cell frequencies of ≥ 0.02%). EMA, ethidium monoazide.