title: Soluble thrombomodulin is antithrombotic in the presence of neutralising antibodies to protein C and reduces circulating activated protein C levels in primates.
abstract: We studied whether there was a relationship between the anticoagulant effects of recombinant human soluble thrombomodulin (rhsTM) and activation of protein C in a primate model of acute vascular graft thrombosis in 11 baboons (Papio species). Baboons were pretreated with 0.1, 1 and 5 mg/kg of rhsTM, with or without co-injection of a neutralising monoclonal antibody to protein C (HPC4) in the 1 mg/kg rhsTM group. Subsequently, thrombogenic polyester grafts were deployed for 3 h into chronic exteriorised arteriovenous shunts. Thrombus growth in the graft, plasma-activated protein C (APC) levels, coagulation and thrombosis markers were determined. In untreated baboons, baseline circulating APC levels more than doubled and graft thrombi propagated until reaching equilibrium in about 1 h. Treatment with rhsTM reduced thrombus propagation rates, prolonged the clotting and bleeding times, decreased thrombin-antithrombin complex, beta-thromboglobulin and fibrinopeptide A levels, and, surprisingly, also decreased systemic APC levels, in a dose-dependent manner. In the presence of HPC4 antibody to inhibit APC generation, the acute antithrombotic activity of rhsTM on graft thromboses was not attenuated for up to 80 min, but sustained thrombus accumulation was observed over a 180-min period. These findings suggest that, in contrast to the prevailing hypotheses, the primary antithrombotic activity of rhsTM is independent of protein C, at least in this primate model. Direct inhibition of thrombin's prothrombotic activity upon complex formation with rhsTM might explain the molecular mechanism of the observed antithrombotic effect.
authors.affiliation: Department of Anesthesiology, Emory University School of Medicine, Atlanta, GA, USA.
authors.forename: Kenichi A
authors.initials: KA
authors.lastname: Tanaka
authors.forename: José A
authors.initials: JA
authors.lastname: Fernández
authors.forename: Ulla M
authors.initials: UM
authors.lastname: Marzec
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Kelly
authors.forename: Mitsunobu
authors.initials: M
authors.lastname: Mohri
authors.forename: John H
authors.initials: JH
authors.lastname: Griffin
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Hanson
authors.forename: Andras
authors.initials: A
authors.lastname: Gruber
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000925:Anticoagulants / Q000627:therapeutic use*; D000990:Antithrombin III; D001777:Blood Coagulation; D001792:Blood Platelets / Q000187:drug effects / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D005337:Fibrin / Q000378:metabolism; D006083:Graft Occlusion, Vascular / Q000097:blood / Q000517:prevention & control*; D008297:Male; D010215:Papio; D010314:Partial Thromboplastin Time; D010447:Peptide Hydrolases / Q000097:blood; D011486:Protein C / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D011994:Recombinant Proteins / Q000627:therapeutic use; D012995:Solubility; D018180:Thrombomodulin / Q000627:therapeutic use*; D013927:Thrombosis / Q000097:blood / Q000276:immunology / Q000517:prevention & control
chemical_list: D000911:Antibodies, Monoclonal; D000925:Anticoagulants; D011486:Protein C; D011994:Recombinant Proteins; D018180:Thrombomodulin; C046193:antithrombin III-protease complex; D000990:Antithrombin III; D005337:Fibrin; D010447:Peptide Hydrolases