Endothelial TM regulates TF-driven coagulation in inflammatory thrombosis model. (A) Schematic showing experimental design. Endothelialized channels were flow adapted at shear stress of 5 dynes/cm2, stimulated with 1 ng/mL TNF-α for 6 hours, and then treated with antibodies or fusion protein during the first 25 minutes of a 30-minute TNF-α washout period, followed by a 5-minute wash to remove nonspecifically bound protein and, finally, infusion of WB at shear stress of 5 dynes/cm2. (B) Blockade of endothelial TF delays and markedly reduces fibrin deposition (P = .02 for AUC of TNF vs TNF + anti-TF mAb). Graph shows mean ± SEM, with n = 2-3 channels per condition. (C) Blockade of endothelial TM results in significant increase in fibrin deposition (P < .001 for AUC of TNF only vs TNF + TM blockade), whereas treatment of TNF-α–activated endothelium with hTM/R6.5 scFv fusion protein completely eliminates fibrin deposition (P = .03 for AUC of TNF only vs TNF + hTM/R6.5). Graph shows mean ± SEM, with n = 2-3 channels per condition. (D) Representative fluorescent images for panel C show fibrin deposition within endothelialized channels and demonstrate opposing effects of TM inhibition and augmentation.
