Fig. 2.

Epitope variants lead to diminished MHC-I binding. A-E) Decreased thermostability of mutant peptide MHC-I complexes. Negative first derivative of relative fluorescence units (rfu) plotted against increasing temperatures. Curves for wild type peptides are black, mutated peptides are colored. The minimum point of the curves represents the melting temperature of peptide-MHC-I complexes. Dashed lines indicate SD. n=2-3 technical replicates. F) Tetramers featuring mutated peptides are unstable at 37°C. FACS plots showing staining of in vitro expanded PBMCs stained with tetramers containing wild type (top) or mutant (bottom) peptides incubated at 4°C (blue) or 37°C (red).