Extended Data Fig. 4: ZipRs augment CAR T cell antitumor activity against LM7 osteosarcoma without altering antigen specificity.
a,b, Cytotoxicity assay after 24-hour co-culture of LM7 WT (left) or LM7 B7−H3 KO (right) cells with CAR T cells at indicated effector:target cell (E:T) ratios (N = 4 (a), N = 3 (b) biological replicates, mean ± SD, ****p < 0.0001, two-way ANOVA with Tukey’s multiple comparisons test). c, Cytokine production after 24-hour co-culture of LM7 WT (left) or LM7 B7-H3 KO (right) cells with CAR or CAR.Zip2R T cells at a 2:1 E:T measured by multiplex analysis (Left: CAR,CAR.Zip2R: N = 3, mean ± SD, ΔCAR, ΔCAR.Zip2R: N = 2; Right: N = 2, mean, two-way ANOVA with Tukey’s multiple comparisons test). d, Cytokine production after 24-hour co-culture of A549 WT (left) or LM7 WT (right) cells with CAR or CAR.Zip7R T cells at a 2:1 E:T measured by multiplex analysis (N = 2 biological replicates, mean). e, Number of stimulations in 7 day repeat stimulation assay with LM7 WT cells and CAR T cells at 2:1 E:T (N = 4, mean ± SD, *p = 0.0154, two-tailed t-test). f, Fold expansion of three representative donors used in repeat stimulation assays with LM7 WT cells. Data represented in (e). g, Repeat stimulation assay with LM7 B7-H3 KO cells and CAR T cells at 2:1 E:T (N = 4, mean ± SD). h, Stimulations of tumor cell killing in 7-day repeat stimulation assay with LM7 WT cells and CAR T cells at 2:1 E:T (N = 3, mean ± SD, *p = 0.0202, two-tailed t-test). i, Fold expansion of three representative donors used in repeat stimulation assays with LM7 WT cells. Data represented in (h). j, Repeat stimulation assay with LM7 B7-H3 KO cells and CAR T cells at 2:1 E:T (N = 3 biological replicates, mean ± SD).
