title: Tissue factor expressed by adherent cells contributes to hemodialysis-membrane thrombogenicity.
abstract: End-stage renal patients present a high risk of thrombosis and bleeding. Consequently, it is challenging to prevent clotting during hemodialysis. If a contact system induces thrombin generation in the extra corporeal circuit, recent data suggest a role of tissue factor (TF) in hemodialysis-associated thrombosis. Using a method of elution, we collected adhering cells to an acrylonitrile membrane layered by polythyleneimine (AN69-ST). Using optic microscopy and flow cytometry, we observed that adherent cells were mainly constituted by activated polymorphonuclear neutrophils (PMNs). Using a sensitive fluorogenic method of thrombin generation, we found that adhering cells triggered thrombin generation in a TF-dependent manner. We next identified the presence of TF mRNA (Q-PCR) in adhering cells. Using immunofluorescence, we observed the presence of TF in PMNs and of TF-decorated neutrophil extracellular traps (NETs). As TF triggers thrombin generation after binding to serine protease FVIIa, we evaluated the effect of an inactivated human recombinant factor VIIa (hrFVIIai) in a sheep model of hemodialysis (HD). One single bolus of hrFVIIai maintained the full patency of the hemodialysis circuit without any measurable systemic anticoagulant effect. TF is a promising target for preventing thrombosis during HD.
authors.affiliation: Equipe d'accueil 3801 Hémostase et Remodelage Vasculaire Post-Ischémie, Université de Reims, France.
authors.forename: Souad
authors.initials: S
authors.lastname: Lakbakbi
authors.affiliation: Centre Hospitalier Universitaire, Service néphrologie, Reims, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Debrumetz
authors.affiliation: Plateforme Imagerie Cellulaire et Tissulaire, Université de Reims, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Terryn
authors.affiliation: Equipe d'accueil 3801 Hémostase et Remodelage Vasculaire Post-Ischémie, Université de Reims, France.
authors.forename: Jean
authors.initials: J
authors.lastname: Szymezak
authors.affiliation: Centre Hospitalier Universitaire, Service néphrologie, Reims, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Rieu
authors.affiliation: Equipe d'accueil 3801 Hémostase et Remodelage Vasculaire Post-Ischémie, Université de Reims, France. Electronic address: pnguyen@chu-reims.fr.
authors.forename: Philippe
authors.initials: P
authors.lastname: Nguyen
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D015942:Factor VIIa / Q000627:therapeutic use; D005260:Female; D008567:Membranes, Artificial; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology*; D011994:Recombinant Proteins / Q000627:therapeutic use; D006435:Renal Dialysis / Q000009:adverse effects* / Q000295:instrumentation; D012756:Sheep; D013917:Thrombin / Q000276:immunology; D013925:Thromboplastin / Q000276:immunology*; D013927:Thrombosis / Q000209:etiology* / Q000276:immunology / Q000517:prevention & control
chemical_list: D008567:Membranes, Artificial; D011994:Recombinant Proteins; D013925:Thromboplastin; C103587:recombinant FVIIa; D015942:Factor VIIa; D013917:Thrombin
keywords: Anticoagulation; Cell activation; Hemodialysis; Tissue factor