Fig. 7: Higher CAR binder B7H3 avidity is associated with increased CD8± CAR-T / tumoroid contact duration and more rapid cytotoxicity against B7H3ultra-dim tumoroids. | Nature Communications

Fig. 7: Higher CAR binder B7H3 avidity is associated with increased CD8± CAR-T / tumoroid contact duration and more rapid cytotoxicity against B7H3ultra-dim tumoroids.

From: Functional avidity of anti-B7H3 CAR-T constructs predicts antigen density thresholds for triggering effector function

Fig. 7

a, b experimental overview- images created with BioRender. (Barisa, M. (2025) https://BioRender.com/ub1tswj). c Illustrative stills of tumoroid and T cell clusters at the start (day 0 = t0), and end (day 7 = t1) 800 min imaging sessions. d CD8+ CAR-T cell cumulative contact time with 691-B and 691-T organoids was tracked for the first 2 h of the first tumor challenge on day 0. The line represents the mean cumulative tumoroid contact score of all the individual CAR-T cells that were successfully tracked for the full 120 min period (the number of cells tracked in each of the conditions was a mean 31.33 ± 10.8; curves were compared using simple linear regressions). e CAR-T cells were incubated with tumoroids overnight at a 1:10 E:T ratio. “Specific cytotoxicity” denotes tumor luminescence relative to matched non-transduced T cell (NTD) controls (mean ± SEM, N = 3 independent experimental replicates, Kruskal–Wallis test). f–i Respective parameters were tracked for the entire duration (800 min) of the first tumor challenge at day 0. f CD4+ and CD8+ CAR-T cell cumulative contact time with 691-B and 691-T organoids (mean ± SEM, N = 31.5 ± 10.6 cells per T cell condition, Two-Way ANOVA). g Cumulative tumoroid death (conversion from viability dye yellow to red) (a total of 398 incremental timepoints were recorded to track tumoroid apoptosis over the assay; curve comparison was carried out using a Friedman test for P value and rank sum differences (RSD)). h CD4+ and CD8+ CAR-T displacement (mean ± SEM, N = 31.5 ± 10.6 cells per T cell condition, two-way ANOVA). i Mean speed of CD4+ and CD8+ CAR-T displacement (mean ± SEM, N = 31.5 ± 10.6 cells per T cell condition, two-way ANOVA). j Trends in CAR-T behavior reduced into six clusters using UMAP analysis of behaviors across all the conditions observed in the initial tumor challenge at day 0 and represented as a heatmap. k The CAR-T behaviors are broken down by T cell, binder and target type. l Pooled data from all CAR constructs against both targets, comparing day 0 and day 7 challenge rate of organoid death.

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