Fig. 2
From: Cancer targeting by TCR gene-engineered T cells directed against Kita-Kyushu Lung Cancer Antigen-1
KK-LC-1 TCR-Ts did not demonstrate cross-reactivity with peptides derived from other human proteins. The IFN-γ production assays shown were performed by coculture of KK-LC-1 TCR-Ts with autologous EBV-LCLs loaded with 1 μg/mL of the peptide indicated. Coculture supernatants were harvested after overnight coincubation. IFN-γ concentration was determined by ELISA. Error bars represent the SD of 2 technical replicates. The “no peptide” conditions had target cells without peptide. “PMA/Iono” indicates T cells that were stimulated with PMA and ionomycin. “UT-Ts” were untransduced control T cells from the same donor as the KK-LC-1 TCR-Ts. a To guide cross-reactivity testing, alanine scanning of KK-LC-152-60 was performed. An alanine residue was substituted for the native residue at each position of KK-LC-152-60. b To compliment alanine substitution and assess the influence of position 7 on target recognition, glycine scanning also was performed. c Peptides derived from human proteins that demonstrated identity at the contact residues inferred by the experiments in (a) and (b) or by a BLAST search for candidate peptides that shared at least 5/9 residues (55% identity) were tested for KK-LC-1 TCR-T recognition