Fig. 4

Experimental validation of T cell response. TIL culture of patient 1 recognized 3 neoantigens, but not the corresponding wildtype peptides. Following pulsing with 10 μg/ml of 25-mer mutant or wt peptide overnight, EBV-transformed autologous B cells B-LCL were co-cultured with T-cells from TIL culture from patient 1. 16 h after the beginning of the co-culture, these cells were co-stained for CD137 (41BB) and CD8+ and analyzed by flow cytometry. The double positive population is indicated in quadrant Q2