biostudies-literatureCai LAugusta UniversityNCI NIH HHSNational Institutes of Health607https://www.ebi.ac.uk/biostudies/studies/S-EPMC7196607biostudies-literatureUnknown11Autologous T cells engineered with T receptor genes (TCR) are being studied to treat cancers. We have recently identified a panel of mouse TCRs specific for the HLA-A0201/alpha fetoprotein epitope (AFP₁₅₈) complex and have shown that human T cells engineered with these TCR genes (TCR-Ts) can eradicate hepatocellular carcinoma (HCC) xenografts in NSG mice. However, due to TCR's promiscuity, their off-target cross-reactivity must be studied prior to conducting clinical trials. In this study, we conducted <i>in vitro</i> X-scan assay and <i>in silico</i> analysis to determine the off-target cross-reactivity of 3 AFP₁₅₈-specific TCR-Ts. We found that the 3 AFP₁₅₈-specific TCR-Ts could be cross-activated by ENPP1₄₃₆ peptide and that the TCR3-Ts could Frontiers in immunologyPreclinical Studies of the Off-Target Reactivity of AFP₁₅₈-Specific TCR Engineered T Cells.PMC7196607R01 CA235159Caraballo Galva LDPeng YYao YHe YCai LLuo XZhu WJi YfalsePreclinical Studies of the Off-Target Reactivity of AFP₁₅₈-Specific TCR Engineered T Cells.Autologous T cells engineered with T receptor genes (TCR) are being studied to treat cancers. We have recently identified a panel of mouse TCRs specific for the HLA-A0201/alpha fetoprotein epitope (AFP₁₅₈) complex and have shown that human T cells engineered with these TCR genes (TCR-Ts) can eradicate hepatocellular carcinoma (HCC) xenografts in NSG mice. However, due to TCR's promiscuity, their off-target cross-reactivity must be studied prior to conducting clinical trials. In this study, we conducted <i>in vitro</i> X-scan assay and <i>in silico</i> analysis to determine the off-target cross-reactivity of 3 AFP₁₅₈-specific TCR-Ts. We found that the 3 AFP₁₅₈-specific TCR-Ts could be cross-activated by ENPP1₄₃₆ peptide and that the TCR3-Ts could 2020-01-01T00:00:00Z20202026-04-18T07:29:18.407Z2020-05-22T19:58:29ZS-EPMC71966073239511710.3389/fimmu.2020.00607