Fig. 1

Design and generation of MSLN-CAR T cells without and with A2aR knockdown. a Schematic representation of four types of second generation of CAR constructs. MSLN-CAR includes anti-mesothelin (MSLN) scFv, human CD8α-derived hinge, 4-1BB derived transmembrane (TM) domain and two intracellular domains including 4-1BB and CD3ζ. Three A2aR-KD1, 2 and 3-MSLN-CARs included all domains of MSLN-CAR plus an anti A2aR shRNA sequence after CD3ζ sequence. shRNA sequences were designed to target different segments of A2aR gene. b Flowcytometry histograms show the percentage of CAR expressing cells 4 days after transduction with the indicated MSLN-CAR-encoding lentiviral vectors at a MOI of ~ 7. The percentage of CAR positive cells is shown. MSLN-CAR: fully human anti mesothelin CAR, A2aR-KD-MSLN-CARs: A2aR-knocked down anti mesothelin-CAR; MOI: multiplicity of infection