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. Author manuscript; available in PMC: 2024 Nov 25.
Published in final edited form as: Nat Biomed Eng. 2023 Nov 30;8(4):380–396. doi: 10.1038/s41551-023-01143-w

Figure 3: ZipRs improve CAR T cell effector function during chronic antigen exposure in vitro.

Figure 3:

(a-b) Transduction efficiency of B7-H3-CAR and Zip2R (a) or Zip7R (b) in human T cells as measured by flow analysis for the ZipR (mClover) and CAR (anti-human F(ab’)2) (N=3-6, mean±SD). (c-d) MTS assay after 24-hour co-culture of A549 WT (left) or A549 B7-H3 KO (right) cells with CAR T cells at indicated effector:target cell (E:T) ratios (N=3-4 biological replicates, mean±SD, ****p<0.0001, two-way ANOVA with Tukey’s multiple comparisons test). (e) Cytokine production after 24-hour co-culture of A549 WT (left) or A549 B7-H3 KO (right) cells with CAR or CAR.Zip2R T cells at a 2:1 E:T measured by multiplex analysis (N=2-3 biological replicates, mean±SD). (f) Stimulations of tumor cell killing in 7-day repeat simulation assay with A549 WT cells and CAR T cells at 2:1 E:T ratio (N=6 biological replicates, mean±SD, ***p<0.001, paired t-test). (g) Three representative donors used for repeat simulation assay with A549 WT cells represented in (f). (h) Repeat stimulation assay with A549 B7-H3 KO cells and CAR T cells at 2:1 E:T (N=4, mean±SD). (i) Stimulations of tumor cell killing in 7 day repeat stimulation assay with A549 WT cells and CAR T cells at 2:1 E:T (N=3 biological replicates, mean±SD, *p<0.05, paired t-test). (j) Three representative donors used for repeat stimulation assay with A549 WT cells represented in (i). (k) Repeat stimulation assay with A549 B7-H3 KO cells and CAR T cells at 2:1 E:T (N=3, mean±SD).