Fig. 4: Joint efficacy–safety analysis of TCB effects reveals differences in immune-cell engagement with healthy and tumour organoids.
a, Left: comparison of cellular architecture and target antigen expression between organoids and tumouroids by H&E and IHC at ×20. Right: quantification of target antigen expression by positive area for each antigen per individual organoid (nOrgCEA = 68, nTumCEA = 76; nOrgEpCAM = 60, nTumEpCAM = 85). ROUT outlier analysis was performed with a Q coefficient of 2%. Statistical analysis was performed using one-way ANOVA. Red line displays mean. Scale for both H&E and IHC, 100 µm. b, Representative single tiles (×20 magnification) of the 7-plex mIF images across all TCB treatments (10 µg ml−1) at 72 h, displaying panCK+ organoids and tumouroids (magenta) surrounded by CD4+ (orange), CD8+ (turquoise), CD14+ (red) and CD20+ (yellow) immune cells. Caspase-3 (green) captures TCB-triggered immune-induced apoptosis, nuclei are stained with DAPI (blue). Scale bar, 100 µm. c, Sum of panCK+ (grey) and caspase-3+ (green) epithelium of the organoids and tumouroids, respectively, detected in zone 2 (on epithelium) across the different TCB treatments and time (n = 3). d, Quantification of the 7-plex mIF images represented in b and e. Heat map of the absolute counts of T-cell subsets within the different zones of individual organoids and tumouroids across time and TCB treatment. e, Single tiles of the 7-plex mIF staining at ×20 magnification (colours explained in b) highlights substantial immune-intercalation dissimilarities between healthy and cancerous epithelium treated with EpCAM TCB (10 µg ml−1). Organoid image: highly CD4+ and CD8+ T-cell infiltrated organoids 24 h post administration. Tumouroid images: progressively killed tumouroid (caspase-3+ apoptotic bodies) over time, but devoid of T lymphocytes within the inner core of the tumouroid. Scale bar, 100 µm. All displayed experiments in this figure were replicated at least three times, yielding similar results.
