Extended Data Fig. 4: Assessment of different hCSF1R CAR constructs in vitro.

(a) Construct design of all anti-human constructs used throughout the course of the study. (b) Representative flow cytometric images of construct expression on primary human T cells. (c) Activation of different hCSF1R CART after incubation with plate-bound hCSF1R protein quantified by flow cytometry. (d) T cells expressing different hCSF1R CAR constructs were cocultured with luc+ target antigen expressing AML tumor cell lines or antigen negative NALM-6 control cells for 48 hours at the indicated E:T ratios. Cell lysis was quantified by BLI. (e) Proliferation dye-labeled hCSF1R CART were cocultured with respective cell lines for 4 or 7 days at a E:T ratio of 0.5:1. Proliferation was subsequently assessed by trace dilution. One representative image of three different donors is shown. (f) Bead quantified T cell numbers. (g) Secretion of IFNγ by T cells transduced with different hCSF1R CAR constructs after co-culture with AML cell lines. (h) Secretion of IFNγ (left) or IL-2 (right) of hCSF1R, CD86 or control CART in co-culture with AML cell lines. (c, d, f - h) Data are mean ± s.e.m. of three independent donors. LTR, long terminal repeat; scFv, single chain fragment variable; TM, transmembrane, IC, intracellular, ED, extracellular domain, CTRL-transduced, control-transduced.