Extended Data Fig. 5: Extended characterization of 80BB and CTLA4 interactions.
From: Synthetic dual co-stimulation increases the potency of HIT and TCR-targeted cell therapies
a, Flow cytometry plots of CTLA4-Fc binding assay. Cells were incubated with increasing concentrations of CTLA4-Fc, and then stained with PE-anti-Fc. b, Ratio of human IL-2 and IL-13 secreted by 19-HIT and 19-HIT80BB cells after 24 hr co-culture with Nalm6CD28 relative to Nalm6. n = 4 independent co-cultures of cells from a healthy donor, representative of 3 donors. c, Serial in vitro cytotoxicity assay using NALM6 or NALM6ΔCTLA4. Fresh target cells were added every 2 days. Each line is an independent co-culture of cells from a healthy donor, representative of 3 donors. d, Left, trace from inference of CRISPR Edits (ICE) tool (Synthego) highlighting extent of CTLA4 knockout. Right, summary of extent of CTLA4 editing indicating CTLA4 knockout evaluated by ICE from 9 donors. e, Serial in vitro cytotoxicity assay of 19-HIT, 19-HIT80ΔBB, or 19-HIT80BB with TRBC, CD28 or CTLA4 genes CRISPR edited. Fresh NALM6 target cells were added every 2 days. Each line is an independent co-culture of cells from a healthy donor, representative of 3 donors. P values were determined by two-tailed t-test (b). Data are mean ± sem.
