Cytokines and growth factors inhibit tumor necrosis factor alpha–induced up-regulation of fibronectin binding on bovine endometrial cells

ObjectiveTo design a high-throughput cell assay to identify molecules modulating adhesion induced by tumor necrosis factor alpha (TNF-α) of endometrial cells to mesothelium.DesignProspective study.SettingBiotech company.Patient(s)Bovine endometrial (BEND) and human mesothelial cells.Intervention(s)Endometrial cells were treated with TNF-α and different proteins.Main Outcome Measure(s)TNF-α increased binding of fibronectin-coated fluorescein isothiocyanate (FITC) beads. The ability of various proteins to inhibit TNF-α–induced fibronectin-bead binding was measured.Result(s)Treatment of BEND cells with TNF-α increased binding of fibronectin-coated beads. Addition of TNF-α–binding protein abrogated the effect of TNF-α in a dose-dependent manner. The initial screen of 1014 proteins identified interferon-α2 (IFN-α2), inteleukin-17 (IL-17), transforming growth factor beta (TGF-β), and platelet-derived growth factor (PDGF) as inhibiting TNF–α-induced bead binding. Interferon-α2, IL-17, and TGF-β inhibited bead-binding with an IC50 (ng/mL, mean ± SD) of 0.15 ± 0.11, 0.098 ± 0.008, and 5.91 ± 0.72, respectively. All three isoforms of PDGF (AA, AB, and BB) were also found to inhibit TNF-α–induced bead-binding, with IC50s (ng/mL) of 1.8 ± 0.45, 10.0 ± 1.49, and 1.72 ± 0.73, respectively.Conclusion(s)We describe a novel high-throughput cell-based assay for endometrial cell binding to fibronectin. We show that IFN-α, IL-17, TGF-β, and PDGF have inhibitory actions on adhesion of endometrial cells to fibronectin.