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. 2011 Aug 12;22(12):1575–1586. doi: 10.1089/hum.2011.070

FIG. 2.

FIG. 2.

Potentially tunable transgene expression and effector functions of RNA CAR+ T cells. (A) Transgene expression of RNA CAR+ T cells electroporated with the indicated amounts of CD19-BBz RNA plotted as a function of time. Histograms of transgene expression of electroporated 19-BBz CAR mRNA are shown. (B) Transgene expression data from (A) plotted as a line graph. Rate of decline is similar despite different MFI seen in (A). (C) Specific lysis of CD19+ tumor cells with CAR T cells electroporated with the indicated amounts of RNA. Lysis was measured by a flow-cytometric CTL assay using K562-CD19 as targets on day 1 (left panel) and day 3 (right panel) after electroporation. Although little difference exists on day 1, by day 3 a dose-dependent decrease in specific lysis relative to RNA dose is observed. (D) IFN-γ secretion by CAR T cells (4 hr after electroporation) with the indicated amounts of RNA cocultured overnight with serially diluted target cells (K562‐CD19) or control targets (K562-meso at 1:1) assayed by ELISA. IFN-γ secretion titrates with the amount of target, and is not statistically significantly different among CD19 CAR+ groups, although a trend toward lower cytokine secretion with lower RNA doses is suggested.