Fig. 4. No cross-reactivity of 10F04mc TCR-T cells.
a Alanine-scan screening to identify 10F04mc recognized key motif. A panel of synthetic peptides containing alanine substitution at all positions in the index HPV18E788-98 peptide were pulsed with K562-DR0901 cells. 10F04mc TCR-T cells were incubated with peptide pulsed K562-DR0901 cells for 16–24 h and IFNγ secretion was detected by ELISA. The alanine mutated position was considered as a key recognition site when the IFNγ secretion induced by this alanine mutated peptide was less than 30% of HPV18E788-98. The letters marked in red represent the mutated amino acids. b 10F04mc transduced T cells were tested by ELISA-IFNγ for recognition of 6 human peptides, which were identified by BLAST since their amino acids sequences were in accord with the pattern of the key motif. The letters highlighted in red represent amino acids that are different from the key epitope. c HLA cross-reactivity analysis of 10F04mc TCR-T cells. 10F04mc transduced T cells were stimulated by LCLs with population representative HLA types pulsed with HPV18E788-98 peptide or without peptide. IFNγ secretion was detected by ELISA. HLA-DRA/DRB1*09:01 positive, n = 4 LCLs; HLA-DRA/DRB1*09:01 negative, n = 12 LCLs. The HLA typing information of all LCLs is shown in Supplementary Table 1. Data are shown as the mean ± SD (a–c), n = 3 biologically independent samples for different groups (a, b). Source data are provided as a Source Data file.