Engineering TCRvβ CARs into healthy T cells results in target elimination. (A) Schematic demonstrating the elimination of a target TCRvβ family (red) within the bulk of healthy donor T cells after transduction with a TCRvβ-targeting CAR and expansion. (B) Representative TCRvβ repertoire of TCRvβ12-CARTs or NTD T cells after expansion determined by flow cytometry. Target population identified with gray box. (C) Targeted (TCRvβ12) and nontargeted (TCRvβ9) surface expression in TCRvβ12-CARTs and NTD T cells after expansion across multiple healthy donors determined by flow cytometry (n = 6) (∗∗∗∗P < .0001 by 2-way ANOVA with Sídák multiple comparison test). (D) Relative surface expression of TCRvβ family members compared with NTD control T cells after transduction with CARs targeting various TCRvβ families. (E) Representative interferon γ (IFN-γ) production by T cells with their full repertoire or sorted to remove the TCRvβ12 population, stimulated with peptides of common viruses, cytomegalovirus (CMV), Epstein-Barr virus (EBV), and influenza virus, and analyzed for production of IFN-γ by ELISpot (not significant [ns] by 2-way ANOVA with Sídák multiple comparison test).