FIG. 1.

Assembly of viral vectors and preparation of effectors for the killing assays. (A) Schematic diagram of the anti-SIV ENV CAR and the control EGFR lentiviral vectors (left) and of the expressed proteins at the cell surface (right). (B) Flow cytometry analysis of CAR T cells for the expression of EGFRt in CD8 (top) and CD4 (bottom) T cells transduced with the SIV ENV CAR or EGFR lentiviral vectors shown in (A). Assessment of the CD8 and CD4 T cell preparations (left) and of EGFRt expression in cells transduced with the EGFR lentivirus (middle) or SIV ENV CAR lentivirus (right). (C) Env binding to CD8 CAR T cells. CD8 CAR T cells or CD8 EGFR T cells were incubated with SIVmac239 gp140 proteins followed by incubation with biotinylated anti-Env mAb and BV421-streptavidin. CAR, chimeric antigen receptor; EGFR, epidermal growth factor receptor; mAb, monoclonal antibody; SIV, simian immunodeficiency virus.