Fig. 1.

Design of CARs and expression in primary human T cells. a Schematic diagram of the lentiviral vector encoding the CARs used in this study. The extracellular domain is tethered to 41BB and the T cell receptor subunit ζ through a transmembrane domain (TM). A T2A peptide sequence separates the CAR from the fluorescent protein BFP. b–c Surface expression of CARs and total BFP expression in primary human T cells. PBMCs were transduced with lentiviral vectors encoding CARs and expanded in the presence of IL-2 (100 U mL⁻¹). The transduction efficiency was determined by the percentage of BFP⁺ cells and the presence of the FLAG-tag on the cell surface as analysed by flow cytometry. d Quantification of the surface expression levels of the five CARs in BFP⁺ cells from three independent healthy donors. Representative data of three independent experiments are shown in b-c