Highly specific lysis of RHAMM/HMMR-R3–, PRAME-P3–, and G250/CA9-G2–pulsed T2 cells in Cr-51 release assays. (A-C) CD8⁺ T cells stimulated with RHAMM/HMMR-R3 (A), PRAME-P3 (B), and G250/CA9-G2 (C) peptides were able to lyse TAA–peptide pulsed T2 cells and primary AML blasts with expression of the respective TAAs (+). In contrast, primary AML blasts not expressing the respective TAAs (–) and K562 cells expressing all TAAs of interest (RHAMM, PRAME, and G250), but lacking expression of HLA molecules, such as HLA-A2, were not lysed by specific CD8⁺ T cells. In accordance, T2 cells pulsed with an irrelevant peptide (MAGE3) were not recognized, underlining the TAA specificity of the T-cell immune response (A-C). HLA-ABC–blocked T2 cells pulsed with G250-G2 peptide were not lysed, but pulsed T2 cells treated with isotype control antibody could be recognized by stimulated CD8⁺ T cells (C).