Fig. 2. Reactivity of B10 CAR-T cells against peptide-pulsed antigen-presenting cells.
a, b Flow cytometric CD107a expression and IFN-γ production analysis of CAR-T cells against T2-A24 cells pulsed with the indicated peptide (a). The gating strategy is shown (b). c LDH release cytotoxicity assay. The results are calculated as %cytotoxicity compared with lysis buffer. The B10 CAR-T cells were co-cultured with peptide-loaded T2-A24 cells at 20 µM. d DNAJB8_143 titration assay. B10 CAR-T cells were co-cultured with T2-A24 cells that were pulsed with a 10-fold serial dilution of DNAJB8_143 peptide (from 10−4 to 10−14 M). The concentrations of IFN-γ in the supernatant are shown. The half-maximal effective concentration (EC50) was calculated by fitting an Emax model. e DNAJB8 alanine scanning. The IFN-γ levels in the supernatant of the culture medium were measured when the B10 CAR-T cells were co-cultured with T2-A24 cells pulsed with DNAJB8_143 peptide or DNAJB8_143 peptides in which one of the amino acids was replaced by alanine, although alanine was itself replaced by serine.