Evaluation of chimeric antigen receptor of humanized rabbit‐derived T cell receptor‐like antibody

T-cell receptor (TCR)-like antibodies that specifically recognize antigenic peptides presented on major histocompatibility complex (MHC) molecules have been developed for next-generation cancer immunotherapy. Recently, we reported a rapid and efficient method to generate TCR-like antibodies using a rabbit system. We humanized previously generated rabbit-derived TCR-like antibodies reacting Epstein-Barr virus peptide (BRLF1p, TYPVLEEMF) in the context of HLA-A24 molecules, produced CAR-T cells, and evaluated their anti-tumor effects using in vitro and in vivo tumor models. Humanization of the rabbit-derived TCR-like antibodies using the complementarity-determining region grafting technology maintained their specificity and affinity. We prepared a second-generation chimeric antigen receptor (CAR) using scFv of the humanized TCR-like antibodies and then transduced them into human T-cells. The CAR-T cells specifically recognized BRLF1p/MHC molecules and lysed the target cells in an antigen-specific manner in vitro. They also demonstrated anti-tumor activity in a mouse xenograft model. We report the generation of CAR-T cells using humanized rabbit-derived TCR-like antibodies. Together with our established and efficient generation procedure for TCR-like antibodies using rabbits, our platform for the clinical application of humanized rabbit-derived TCR-like antibodies to CAR-T cells will help improve next-generation cancer immunotherapy.