Fig. 6: Limited efficacy of etrolizumab surrogate (ETZ-s) to block E-Cadherin-dependent co-stimulation of CD8⁺ T cells.

A Schematic depiction of the experimental setup in the co-stimulation assays. Magnetically enriched peripheral blood CD8⁺ T cells were stimulated for 3 days with anti-CD2/3/28 beads in the presence of 10 ng/ml TGFβ1, rested for 1 day, and subsequently re-stimulated on plates coated with either 1 or 5 µg/ml anti-CD3 together with or without (w/o) E-Cadherin (E-Cad) in the presence of 10 µg/ml ETZ-s or an isotype control (Iso). Prepared with licensed BioRender application. B Representative and quantitative flow cytometry of CD69 expression after re-stimulation. Significant outliers were detected by 1% ROUT test and removed from analysis. Normality was determined by D’Agostino and Pearson test and repeated measures one-way ANOVAs with Dunnett’s multiple comparisons test were performed accordingly. Change of CD69 expression compared to w/o + Iso (C) and change of CD69 expression compared to E-Cad + Iso (D). Significant outliers were detected by 1% ROUT test and removed from analysis. Normality was determined by D’Agostino and Pearson test and two-tailed paired and one sample t tests were performed accordingly. n = 12. E Relative concentrations of the indicated proteins in the supernatants of re-stimulated cultures relative to E-Cad + Iso. Significant outliers were detected by 1% ROUT test and removed from analysis. Two-sided one sample Wilcoxon signed rank tests were performed. n = 8–11. Data are displayed as box-whisker plots from minimum to maximum. The center line indicates the median with box limits defining the quartiles. Source data are provided as a Source Data file. TCR T cell receptor.