Domain-swapped T cell receptors improve the safety of TCR gene therapy


  • M.T. Bethune
  • M.H. Gee
  • M. Bunse
  • M.S. Lee
  • E.H. Gschweng
  • M.S. Pagadala
  • J. Zhou
  • D. Cheng
  • J.R. Heath
  • D.B. Kohn
  • M.S. Kuhns
  • W. Uckert
  • D. Baltimore


  • eLife


  • eLife 5: e19095


  • T cells engineered to express a tumor-specific {alpha}{beta} T cell receptor (TCR) mediate anti-tumor immunity. However, mispairing of the therapeutic {alpha}{beta} chains with endogenous {alpha}{beta} chains reduces therapeutic TCR surface expression and generates self-reactive TCRs. We report a general strategy to prevent TCR mispairing: swapping constant domains between the {alpha} and {beta} chains of a therapeutic TCR. When paired, domain-swapped (ds)TCRs assemble with CD3, express on the cell surface, and mediate antigen-specific T cell responses. By contrast, dsTCR chains mispaired with endogenous chains cannot properly assemble with CD3 or signal, preventing autoimmunity. We validate this approach in cell-based assays and in a mouse model of TCR gene transfer-induced graft-versus-host disease. We also validate a related approach whereby replacement of {alpha}{beta} TCR domains with corresponding {gamma}{delta} TCR domains yields a functional TCR that does not mispair. This work enables the design of safer TCR gene therapies for cancer immunotherapy.