1. EnAd‐EpCAMBiTE shows reproducible activity within an expanded cohort of patient malignant peritoneal and pleural exudates
EnAd‐EpCAMBiTE shows reproducible activity within an expanded cohort of patient malignant peritoneal and pleural exudates A–CUnpurified total cells from ascites or pleural effusions (from seven different patients; pleural effusion, blue; peritoneal ascites, red) were incubated in 50% fluid from the same exudate sample in the presence of free BiTE, EnAd or recombinant virus. After 5 days, the total cell population was harvested, and the number of (A) CD3+ T cells and those which were (B) CD25+ was quantified. (C) The number of EpCAM+ cells was measured using flow cytometry.DRepresentative microscopy images (magnification ×10; scale bar, 100 μm) and flow cytometry analysis of pleural effusion cells of patient 3 (cancer cells and lymphocytes) following treatment with EnAd or EnAd‐SA‐EpCAM BiTE. Pink, total cells; blue, CD3+ cells; red, CD3+CD25+ cells; black, EpCAM+ cells.EAt 5 days, cytokine levels were measured by LEGENDplex human Th cytokine panel using pleural effusion cultures following incubation with free recombinant BiTE or infection with EnAd or recombinant virus. E:T (CD3+:EpCAM+) ratio upon receipt of patient samples were 0.51 (patient 1), 37.6 (patient 2), 0.39 (patient 3), 4.44 (patient 4), 0.14 (patient 5), 6.12 (patient 6) and 30.6 (patient 7).Data information: Each condition was measured in biological triplicate and represented as mean ± SD. Significance was assessed by comparison to untreated control samples using a one‐way ANOVA test with Tukey's post hoc analysis, *P < 0.05, **P < 0.01, ***P <  Source data are available online for this figure.
Joshua D Freedman et al. EMBO Mol Med. 2017;emmm
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