Recognition of LCL-TM cells cannot be outcompeted by peptides. (A) T2 cells were transduced with HLA-A*24:02 and loaded with 10 µM of 15 different transformation-associated peptides (supplemental Table 1), after which they were coincubated with TEG011 cells. Activation of TEG011 cells was assessed by measuring IFN-γ production. (B) The 4 residues of NEF_134-144 that are pointed out of the peptide binding groove of HLA-A*24:02 are indicated. (C) Ten micromoles of the generated NEF_134-144-derived mutant peptides were loaded on LCL-TM cells, after which they were coincubated with TEG011 cells. (D) HLA-A*24:02-transduced K562 cells were loaded with 10 µM NEF_134-144 before loading with increasing concentrations of an HLA-A*24:02-restricted MiHA peptide (K.F., unpublished data). The peptide-loaded loaded cells were coincubated with TEG011 cells, αβT cells engineered with an HLA-A*24:02-restricted NEF_134-144-specific TCR or a MiHA-specific αβT cell clone. T-cell activation was assessed by measuring IFN-γ. Error bars represent the SD (n ≥ 2).