Fig. 5
From: Bi- and tri-valent T cell engagers deplete tumour-associated macrophages in cancer patient samples
A CD206 TriTE with bivalent CD3 binding retains specificity for M2 macrophages and overcomes ascites suppression. a Monocyte-derived macrophages were polarised as indicated, CFSE-stained, and co-cultured for 96 h with T cells at increasing E:T ratios and BiTE/TriTE concentrations. % Live cells were calculated with propidium iodide staining and Celigo image cytometry, with values displayed as a heat map. b T cells were co-cultured with monocyte-derived macrophages and BiTEs/TriTEs in the presence or absence of 50% ascites fluid from seven different patients (Patients 1, 2, 3, 4, 6, 7 and 8). 96 h later, CD25 expression was determined by flow cytometry. C, CFSE-stained monocyte-derived macrophages were treated with T cells (10:1 E:T ratio) and BiTEs/TriTEs in medium alone or 50% ascites supernatant from seven different patients (Patients 1, 2, 3, 4, 6, 7 and 8). 96 h later, cells were stained with proprodium iodide and analysed with a Celigo image cytometer to calculate % live cells. Data show mean ± SD of biological triplicates (b, c). Statistical significance was assessed by two-way ANOVA followed by Bonferroni post-hoc analysis, with each treatment being compared to the relevant “Mock” condition (b, c) (*, P < 0.05; **, P < 0.01; ***, P < 0.001)