Subscribe to RSS
DOI: 10.1055/s-0037-1614240
Antagonism of vWF Inhibits both Injury Induced Arterial and Venous Thrombosis in the Hamster
Publication History
Received
May 1997
Accepted after resubmission
20 August 1997
Publication Date:
08 December 2017 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199801/lookinside/thumbnails/10-1055-s-0037-1614240-1.jpg)
Summary
von Willebrand factor (vWF) is instrumental in arterial but has also been implicated in venous thrombogenesis. To address its role in venous thrombosis, experimental thrombosis was induced in the carotid artery and the femoral vein of hamsters, following which thrombus prevention by two different antagonists of vWF was studied. The first antagonist was the anti-human vWF monoclonal antibody AJvW-2, which inhibits the botrocetin and ristocetin induced aggregation of human blood platelets. AJvW-2 reacts with an epitope present in the A1 domain of vWF in very different species (human, pig, rabbit, dog, Guinea pig and rat). This epitope was found to be conformational and overlapping with vWF binding sites for aurin tricarboxylic acid (ATA), but not for botrocetin and heparin. AJvW-2 has affinities for vWF in the absence (Kd = 0.5 ± 0.03 nmol/l in solution) and in the presence of shear stress (Kd = 3.3 ± 0.6 nmol/l during perfusion at 1,300 s–1 over subendothelial matrix associated vWF) sufficiently elevated to neutralize vWF. During perfusion of subendothelial matrix with anticoagulated human blood, the surface covered by adhering platelets was reduced by AJvW-2, with IC50s equal to 6.6 ± 0.34 μg/ml at 1,300 s–1 and to 1 ± 0.01 μg/ml at 2,700 s-1. As a second antagonist, molecular size gel filtered ATA was selected. Fractionated ATA inhibited platelet adhesion to matrix with IC50s equal to 0.27 ± 0.09 mmol/l at 1,300 s –1 and 0.16 ± 0.008 mmol/l at 2,700 s –1. When administered to hamsters, AJvW-2 prevented thrombosis in the injured carotid artery dose-dependently (ED50 = 0.15 ± 0.01 mg/kg). Thrombosis in the similarly injured femoral vein was however also inhibited (ED50 = 0.37 ± 0.06 mg/kg). Likewise, fractionated ATA completely inhibited carotid artery thrombosis (ED50 = 0.42 ± 0.13 mg/kg), but also interfered with femoral vein thrombosis (apparent ED50 between 2 and 3 mg/kg). We conclude that antagonizing the vWF A1 domain by AJvW-2 and to a lesser extent also by fractionated ATA, inhibits thrombosis not only in the arterial but also in the venous circulation. Since venous thrombi were prevented at only 3-5-fold higher doses of antagonist, vWF participates in injury induced venous thrombosis.
-
References
- 1 Sadler JE. von Willebrand factor. J Biol Chem 1991; 266: 22777-80.
- 2 Rand JH, Wu XX, Potter BJ, Uson RR, Gordon RE. Co-localization of von Willebrand factor and type VI collagen in human vascular subendothelium. Am J Pathol 1993; 142: 843-50.
- 3 Girma JP, Kalafatis M, Pietu G, Lavergne JM, Chopek MW, Edgington TS, Meyer D. Mapping of distinct von Willebrand factor domains interacting with platelet GPIb and GPIIb/IIIa and with collagen using monoclonal antibodies. Blood 1986; 67: 1356-66.
- 4 Berndt MC, Ward CM, Booth WJ, Castaldi PA, Mazurov AV, Andrews RK. Identification of aspartic acid 514 through glutamic acid 542 as a glycoprotein Ib-IX complex receptor recognition sequence in von Willebrand factor. Mechanism of modulation of von Willebrand factor by ristocetin and botrocetin. Biochemistry 1992; 31: 11144-51.
- 5 Cruz MA, Yuan H, Lee JR, Wise RJ, Handin RI. Interaction of the von Willebrand factor (vWF) with collagen. Localization of the primary collagen-binding site by analysis of recombinant vWF a domain polypeptides. J Biol Chem 1995; 270: 10822-7.
- 6 Lankhof H, van Hoeij M, Schiphorst ME, Bracke M, Wu YP, Ijsseldijk MJ, Vink T, de Groot PG, Sixma JJ. A3 domain is essential for interaction of von Willebrand factor with collagen type III. Thromb Haemost 1996; 75: 950-8.
- 7 Sakariassen KS, Bolhuis PA, Sixma JJ. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII-Von Willebrand factor bound to the subendothelium. Nature 1979; 279: 636-8.
- 8 Miyata S, Goto S, Federici AB, Ware J, Ruggeri ZM. Conformational changes in the A1 domain of von Willebrand factor modulating the interaction with platelet glycoprotein Ib alpha. J Biol Chem 1996; 271: 9046-53.
- 9 Siedlecki CA, Lestini BJ, Kottke-Marchant K, Eppell SJ, Wilson DL, Mar-chant RE. Shear-dependent changes in the three-dimensional structure of human von Willebrand factor. Blood 1996; 88: 2939-50.
- 10 Nichols TC, Bellinger DA, Reddick RL, Read MS, Koch GG, Brinkhous KM, Griggs TR. Role of von Willebrand factor in arterial thrombosis. Studies in normal and von Willebrand disease pigs. Circulation 1991; 83: IV56-IV64.
- 11 Nichols TC, Samama CM, Bellinger DA, Roussi J, Reddick RL, Bonneau M, Read MS, Bailliart O, Koch GG. Vaiman, et al. Function of von Willebrand factor after bone marrow transplantation between normal and von Willebrand disease pigs: effect on arterial thrombosis in chimeras. Proc Natl Acad Sci 1995; 92: 2455-9.
- 12 Roussi J, Samama M, Vaiman M, Nichols T, Pignaud G, Bonneau M, Sigman J, de Castro H, Griggs T, Drouet L. An experimental model for testing von Willebrand function: successful SLA-matched crossed bone marrow transplantations between normal and von Willebrand pigs. Exp Hematol 1996; 24: 585-91.
- 13 Cadroy Y, Hanson SR, Kelly AB, Marzec UM, Evatt BL, Kunicki TJ, Montgomery RR, Harker LA. Relative antithrombotic effects of monoclonal antibodies targeting different platelet glycoprotein-adhesive molecule interactions in non-human primates. Blood 1994; 83: 3218-24.
- 14 Bellinger DA, Nichols TC, Read MS, Reddick RL, Lamb MA, Brinkhous KM, Evatt BL, Griggs TR. Prevention of occlusive coronary artery thrombosis by a murine monoclonal antibody to porcine von Willebrand factor. Proc Natl Acad Sci USA 1987; 84: 8100-4.
- 15 Golino P, Ragni M, Cirillo P, Pascucci I, Ezekowitz MD, Pawashe A, Scognamiglio A, Pace L, Guarino A, Chiariello M. Aurintricarboxylic acid reduces platelet deposition in stenosed and endothelially injured rabbit carotid arteries more effectively than other antiplatelet interventions. Thromb. Haemost 1995; 74: 974-9.
- 16 Strony J, Song A, Rusterholtz L, Adelman B. Aurintricarboxylic acid prevents acute rethrombosis in a canine model of arterial thrombosis. Arterioscler Thromb Vasc Biol 1995; 15: 359-66.
- 17 Azzam K, Garfinkel LI, Bal-dit-Sollier C, Cisse-Thiam M, Drouet L. Antithrombotic effect of a recombinant von Willebrand factor, VCL, on nitrogen laser-induced thrombus formation in guinea pig mesenteric arteries. Thromb Haemost 1995; 73: 318-23.
- 18 Cho J-S, Ouriel K. Differential thrombogenicity of artery and vein: the role of von Willebrand factor. Ann Vasc Surg 1995; 9: 60-70.
- 19 Bernat A, Lale A, Herbert JM. Aurin tricarboxylic acid inhibits experimental venous thrombosis. Thromb Res 1994; 74: 617-27.
- 20 Yamamoto H, Nagano M, Kitoh M, Kobayashi T, Yoshimoto R. Monoclonal antibody against von Willebrand factor inhibits thrombus formation without prolongation of bleeding time. Blood. 1995 86 Suppl 1: Abstract 325, 84a.
- 21 Kageyama S, Yamamoto H, Nagano M, Arisaka H, Kayahara T, Yoshimoto R. Anti-human von Willebrand factor antibody AJvW-2 inhibits arterial thrombosis with low risk of bleeding. Br J Pharmacol 1997; 122: 165-71.
- 22 Blann AD, McCollum CN. von Willebrand factor, endothelial cell damage and atherosclerosis. Eur. J Vasc Surg 1994; 8: 10-15.
- 23 Weinstein M, Vosburgh E, Philips M, Turner N, Chute-Rose L, Moake J. Isolation from commercial aurintricarboxylic acid of the most effective polymeric inhibitors of von Willebrand factor interaction with platelet glycoprotein Ib. Comparison with other polyanionic and polyaromatic polymers. Blood 1991; 78: 2291-8.
- 24 Stockmans F, Deckmyn H, Gruwez J, Vermylen J, Acland R. Continuous quantitative monitoring of mural, platelet-dependent, thrombus kinetics in the crushed rat femoral vein. Thromb Haemost 1991; 65: 425-31.
- 25 Ruggeri ZM, Zimmerman TS, Russell S, Bader R, De Marco L. von Willebrand factor binding to platelet glycoprotein Ib complex. Methods Enzymol 1992; 215: 263-75.
- 26 Loscalzo J, Handin RI. Platelet glycocalicin. Methods Enzymol 1992; 215: 289-94.
- 27 Hoylaerts MF, Nuyts K, Peerlinck K, Deckmyn H, Vermylen J. Promotion of binding of von Willebrand factor to platelet glycoprotein Ib by dimers of ristocetin. Biochem. J 1995; 306: 453-63.
- 28 Hoylaerts MF, Bollen A, De Broe ME. The application of enzyme kinetics to the determination of dissociation constants for antigen-antibody interactions in solution. J Immunol Meth 1990; 126: 253-61.
- 29 Hoylaerts MF, Yamamoto H, Vreys L, Nuyts K, Deckmyn H, Vermylen J. Binding of von Willebrand factor to collagen VI is primarily mediated via its A1 domain. Bioch J 1997; 324: 185-91.
- 30 Sakariassen KS, Aarts PA, de Groot PG, Houdijk WP, Sixma JJ. A perfusion chamber developed to investigate platelet interaction in flowing blood with human vessel wall cells, their extracellular matrix, and purified components. J Lab Clin Med 1983; 102: 522-35.
- 31 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemost 1987; 58: 1078-84.
- 32 Stockmans F, Stassen JM, Vermylen J, Hoylaerts MF, Nyström A. A tech nique to investigate mural thrombus formation in small arteries and veins: I. Comparative morphometric and histological analysis. Ann Plast Surg 1997; 38: 56-62.
- 33 Stockmans F, Stassen JM, Vermylen J, Hoylaerts MF, Nyström A. A tech nique to investigate mural thrombus formation in small arteries and veins: II. Effects of aspirin, heparin, r-hirudin and G-4120. Ann Plast Surg 1997; 38: 63-8.
- 34 Slack SM, Turitto VT. Flow chambers and their standardization for use in studies of thrombosis. Thromb. Haemost 1994; 72: 777-81.
- 35 Christophe O, Rouault C, Obert B, Pietu G, Meyer D, Girma JP. A monoclonal antibody (B724) to von Willebrand factor recognizing an epitope within the A1 disulphide loop (Cys509-Cys695) discriminates between type 2A and type 2B von Willebrand disease. Br J Haematol 1995; 90: 195-203.
- 36 Kawasaki T, Kaku S, Kohinata T, Sakai Y, Taniuchi Y, Yano S, Takenaka T, Fujimura Y. Inhibition by aurin tricarboxylic acid of von Willebrand factor binding to platelet GPIb, platelet retention and thrombus formation in vivo. Am J Hematol 1994; 47: 6-15.
- 37 Nichols TC, Bellinger DA, Johnson TA, Lamb MA, Griggs TR. von Willebrand’s disease prevents occlusive thrombosis in stenosed and injured porcine coronary arteries. Circ Res 1986; 59: 15-26.
- 38 Nichols TC, Bellinger DA, Reddick RL, Smith SV, Koch GG, Davis K, Sigman J, Brinkhous KM, Griggs TR, Read MS. The roles of von Willebrand factor and factor VIII in arterial thrombosis: studies in canine von Willebrand disease and hemophilia A. Blood 1993; 81: 2644-51.
- 39 d’Alessio P, Zwaginga JJ, de Boer HC, Federici AB, Rodeghiero F, Castaman G, Mariani G, Mannucci PM, de Groot PG, Sixma JJ. Platelet adhesion to collagen in subtypes of type I von Willebrand’s disease is dependent on platelet von Willebrand factor. Thromb. Haemost. 1990; 64: 227-31.
- 40 Turitto VT, Weiss HJ, Baumgartner HR. Platelet interaction with rabbit subendothelium in von Willebrand’s disease: altered thrombus formation distinct from defective platelet adhesion. J. Clin. Invest. 1984; 74: 1730-41.