Fig. 4

Knock-in GD2-CAR-T cells do not show phenotypic differences and are not functionally inferior when compared to viral GD2-CAR-T cells. a Changes of CD4/CD8 ratio after processing of leukopaks and over time. b + c Memory marker (b) and exhaustion marker (c) expression of viral vs. GD2 knock-in CAR-T cells as determined via flow cytometry on day 14 (pooled data from n = 3 independent donors). d GD2 antigen levels of co-cultured tumor cell lines. Representative histograms from n = 3 independent experiments. e Intracellular cytokine (TNF-a, IL-2, IFN-g) and activation marker (CD107a, CD69) expression after 6 h of co-culture with respective GD2 expressing tumor cell lines. Shown here is the marker positive cell frequency gated on CD8 + CAR + T cells (pooled data from n = 2 independent donors tested in technical triplicates). f Concentration dependent tumor cell killing after 48 h of co-culture with indicated E:T ratios (pooled data from n = 3 independent donors). g Tumor cell killing over time in GD2 antigen expressing tumor cell lines at a E:T ratio of 1:10 (pooled data from n = 2 independent donors tested in technical triplicates). Error bars indicate SD