Functional expression of lentivirally transduced 3C10-CAR and miR-17-92 in human T cells. CD3+ T cells were transduced with pELNS-3C10-CAR alone or with both pELNS-3C10-CAR and FG12-EF1a-miR-17/92 as described in Methods. (A) Transduced T-cells were evaluated by flow cytometry for expression of 3C10-CAR and miR-17-92 by anti-mouse (Fab’)2 antibody and EGFP, respectively. (B) Expression levels of the miR-17-92 cluster members miR-17-3p, miR-17-5p, and miR-92a-1 in transduced T cells were measured by qRT-PCR. Mean ± SD values of 3 replicate measurements from one of three experiments with similar results are depicted. * indicates p < 0.05 between the two groups using student t-test. (C) EGFRvIII-specific cytotoxic activities of transduced T cells evaluated by a 12-h 51Cr-release assay at various E:T ratios against 51Cr-labeled U87-EGFRvIII or control U87 cells. Control cells were Mock (EGFP)-transduced T-cells. Values indicate mean ± SD in triplicated wells.