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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: Cancer Immunol Res. 2015 May 4;3(7):815–826. doi: 10.1158/2326-6066.CIR-15-0054

Figure 1. KIR-CARs/Dap12 trigger robust antigen-specific cytotoxicity in vitro.

Figure 1

A) Schematic diagram of a mesothelin-specific KIR–CAR. B) Schematic diagram of the lentiviral vector used for co-delivery of a KIR-CAR and DAP12. C) Primary human T cells were simulated with anti-CD3/anti-CD28 T-cell activator beads (Dynabeads® CD3/CD28 CTS™, Life Technologies). After 24 hrs of stimulation, the T cells were transduced with a lentiviral vector encoding the indicated CAR or mock-transduced (Mock). Cells were expanded for 9 days, and analyzed by flow cytometry for the expression of the indicated CAR using biotinylated goat-anti-mouse F(ab)2 followed by streptavidin-PE. Results are representative of at least 3 independent experiments. D) Antigen-specific cytotoxic activity of T cells generated as described in panel C was assessed by a 4-hr 51Cr-release assay as described in the supplemental materials and methods using K562 cells (Kwt) or K562 cells engineered to express human mesothelin (Kmeso).