Figure 2

Transduction efficacy and purity of CD8⁺T cells (a-c) and functional activity of CAR transduced T cells in vitro (d, e, f). Transduction efficacy was assessed by staining the CH2/3 immunoglobulin linker domain with anti-hIgG mAb and with anti-CD8 mAb (n = 10). A shows non-transduced T cells, whereas b displays T cells transduced with the FAP-specific anti-FAP-F19-∆CD28/CD3ζ CAR and c with the NY-ESO-1 specific anti-NY-ESO-1-T1-∆CD28/CD3ζ CAR. 24 h co-cultivation of re-directed T cells with the target cell lines HT1080FAP-luc and T2-1B was analyzed for IFNγ release (n = 5) (d). Comparison of IL-2 production of anti-FAP-F19-∆CD28/CD3ζ and anti-FAP-F19-CD28/CD3ζ after 24 h of co-cultivation with HT1080FAP-luc (n = 2) (e). Proliferation of anti-FAP-F19-∆CD28/CD3ζ re-directed T cells after stimulation with recombinant human FAP (1 μg/ml) in the absence and presence of IL-2 (100 IU/ml) at indicated time points (n = 2) (f). Error bars were calculated as mean + SD (***p < 0.001, **p < 0.01 as calculated by Student’s unpaired 2 tailed t-tests).