EBV-CTLs generated from patients with HD can be grafted with a functional CD30CAR. EBV-CTL lines were expanded from PBMCs of 4 patients with HD. (A) The expression of CD30CAR on 2 representative CTL lines by flow cytometry using a goat anti–human IgG (H + L) Ab (solid line) is shown. The dotted line shows the isotype control. (B) The immunophenotype of EBV-CTLs generated from these 4 HD patients and transduced with the CD30CAR (■) compared with EBV-CTLs transduced with an irrelevant CAR (□) is shown. Mean and SD are shown. (C) The frequency of tetramers recognizing the lytic (BZLF1-RAK) EBV-associated antigen in EBV-CTLs generated from 1 of these patients is shown. The bottom panels show that the same frequency of EBV-specific tetramers is maintained after transduction with CD30CAR and that the CD30CAR is also detectable on tetramer⁺ T cells. (D) The killing of LCLs and CD30⁺ tumor cell lines in a standard ⁵¹Cr-release assay at a CTL/tumor cell ratio of 20:1 is shown. Bars represent the mean plus or minus the SD of the EBV-CTLs transduced with the CD30CAR (■) or an irrelevant CAR (□). CD30CAR⁺ CTLs lysed both autologous LCLs and CD30⁺ target cells, whereas control CTLs showed significant lysis only of autologous LCLs.