Fig. 2

CD8+ SS1 CAR T-cells have increased pZAP70 after pervanadate stimulation. A CAR scFv distribution in the cell membrane in untransduced cells (UT), Mutζ, CD28ζ and 4-1BBζ CARs; CD3 (Green), CAR scFv (Red), DAPI (Blue). B–E Relative levels of ZAP70 expression by flow cytometry. Histograms of ZAP70 in a representative donor’s B CD4+, and D CD8+ cell products. Bar graphs of ZAP70 expression from three donors measured by MFI in C CD4+, and E CD8+ cell products. F–M Relative levels of phosphorylated ZAP70 (pZAP70) from three donors by flow cytometry. Histograms of pZAP70 in a representative donor’s F CD4+, and H CD8+ cell products at rest and J–L after stimulation with pervanadate. Bar graphs of pZAP70 activation measured by MFI in G CD4+, and I CD8+ cell products at rest, and K, M after stimulation with pervanadate. The modal peak of untransduced T-cells is indicated by the dashed line in the histograms. Bar graphs are of n = 3 independent donors analysed on different days; each donor is distinguished using a unique symbol. Data in bar graphs represent mean ± SEM. Comparisons were made between all cell products by one-way ANOVA with Tukey’s correction for multiple comparisons; only statistically significant differences are indicated (*p < 0.05, **p < 0.01)