Skip to main content
. 2009 Apr 21;17(8):1453–1464. doi: 10.1038/mt.2009.83

Figure 1.

Figure 1

Lentiviral gene transfer combined with αCD3/αCD28 coated magnetic bead activation of T cells permits generation of large numbers of CD19-specific chimeric antigen receptor (CAR+) T cells. (a) A schematic diagram showing the CD19-specific CAR used in this study. (b) Comparison of green fluorescent protein (GFP) expression under the control of different eukaryotic promoters in primary human CD4+ and CD8+ T cells over time. GFP fluorescence was compared in the indicated T cell subset in cells that were stimulated with αCD3/αCD28 coated beads followed by lentiviral transduction at an multiplicity of infection (MOI) of 0.2 on day 1 with vector expressing enhanced GFP under the control of the promoter indicated. Flow cytometric detection of GFP fluorescence was calibrated using Rainbow Calibration Particles (Spherotech, Lake Forest, IL) to correct for day-to-day variation. (c) αCD19-specific CAR surface expression in primary human CD4+ and CD8+ T cells. Expression was examined 6 days following transduction with the indicated CAR-encoding lentiviral vector at a MOI of ~8. (d) In vitro expansion of CD4+ and CD8+ T cells following activation with αCD3/αCD28 coated magnetic beads and transduction of the indicated CAR on day 1. Data are representative of >3 independent experiments.