Figure 3 from Activation of Cell-Intrinsic Signaling in CAR-T Cells via a Chimeric IL7R Domain

CD123-targeted CAR and CCR can be coexpressed with maintained functionality. A, Schematic showing CAR and CCR structures. B, Transduction efficiency measured using flow cytometric staining of Halo and SNAP tags. n = 3 to 6 unique T-cell donors, unless noted no significant difference between groups. C, Measurement of phosphorylated STAT5 percentage in cytokine starved, engineered T cells measured with and without activation on the immobilized rhCD123 target. n = 3 to 4 unique T-cell donors, comparison of CCR + CAR⁺ and CAR⁺ to NT (black) and with stimulation (colored). D, Soluble IL2 and IFNγ measured in the supernatant following coculture of unmodified (NT), CCR⁺, CAR⁺, or CCR + CAR⁺ T cells with CD123-negative (K562) and CD123-positive (K562.CD123, MV-4-11, and Molm-13) targets. n = 3 to 6 unique T-cell donors; data represented as mean ± SD. Significance noted is in comparison to NT (black asterisks) and/or CCR⁺ cells (green asterisks) or as noted. NT vs. CCR⁺ comparison was nonsignificant in all instances. E, Bioluminescence-based cytotoxicity assays performed using K562, K562.CD123, MV-4-11, and Molm-13 stably expressing ffLuc; n = 3 to 5 donors. For B–E, *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. Unless noted, comparisons were nonsignificant. NT, nontransduced.