Figure 3.

Initial comparison of VLR- and scFv-based CD5-CARs. (A) Jurkat T cells were transduced with lentiviral vectors encoding either an scFv- or VLR-based CD5-CAR with co-expression of eGFP. Schematic of the Jurkat T cell activation assay shows time points for measurement of T-cell activation and Western blot analysis. (B) Activation measured by surface CD69 expression four days after transduction increased as the amount of viral vector increased. Greater activation was observed in the CD5-VLR-CAR Jurkat group. (C) The percentage of activated cells was compared to the vector copy number (VCN) obtained for each transduced population of cells. The inset to the figure defines each group. (D) CD69 expression was measured 4 and 12 d after transduction, which showed activation decreased over time in both CD5-CAR expressing Jurkat T cell groups. Errors bars represent standard deviations.