PDF herunterladen
Thromb Haemost 1993; 70(03): 522-526
DOI: 10.1055/s-0038-1649616
Original Article
Platelets
Schattauer GmbH Stuttgart

Platelet Aggregation on Extracellular Matrix: Effect of a Recombinant GPIb-Binding Fragment of von Willebrand Factor

R Dardik
1   National Hemophilia Center, Sheba Medical Center, Tel Hashomer, Israel
,
Z M Ruggeri
2   Roon Research Center for Arteriosclerosis and Thrombosis, Department of Molecular and Experimental Medicine and Committee on Vascular Biology, The Scripps Research Institute, La Jolla, California, USA
,
N Savion
3   Goldschleger Eye Research Institute, Sackler Faculty of Medicine, Tel Aviv University, Israel
,
S Gitel
1   National Hemophilia Center, Sheba Medical Center, Tel Hashomer, Israel
,
U Martinowitz
1   National Hemophilia Center, Sheba Medical Center, Tel Hashomer, Israel
,
V Chu
4   Rhone-Poulenc Rorer Central Research, King of Prussia, Pennsylvania, USA
,
D Varon
1   National Hemophilia Center, Sheba Medical Center, Tel Hashomer, Israel
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received 24. September 1992

Accepted after revision 16. April 1993

Publikationsdatum:
05. Juli 2018 (online)

  als PDF herunterladen  Lizenzen und Reprints

Summary

Platelets in whole blood incubated on extracellular matrix (ECM) produced by bovine corneal endothelial cells under oscillatory flow conditions demonstrate extensive aggregate formation. Since both platelet-subendothelium and platelet-platelet interactions are mediated by von Willebrand factor (vWF), we used this system to examine the effect of a recombinant GPIb-binding fragment of vWF (designated RG12986), comprising residues 445-733 of the native vWF subunit, on platelet reactivity with ECM. The seven cysteines present in the RG12986 fragment were reduced and alkylated in order to achieve a monomeric conformation. The recombinant vWF fragment binds to unstimulated platelets in the absence of exogenous modulators. When added to platelet-rich plasma, it inhibits ristocetin-induced platelet agglutination. Binding of 51Cr-labeled platelets in reconstituted whole blood to ECM was inhibited by RG12986 in a dose dependent and saturable manner, with IC50 of 4 μM and maximal inhibition (about 70%) at 6 μM. Scanning electron microscope (SEM) analysis showed that addition of RG12986 to whole blood significantly inhibited platelet aggregation on ECM. The extent of inhibition observed with RG12986 at a final concentration of 4 μM was similar to that obtained with the cell adhesion peptide RGDS at the concentration of 0.1 mM. The ability of the RG12986 fragment to inhibit platelet aggregation on ECM is in agreement with the concept that blockade of vWF-GPIb interaction may inhibit further events leading to activation of the glycoprotein IIb/IIIa (GPIIb/IIIa) complex and subsequent thrombus formation.

 

  • References

  • 1 George JN, Nurden AT, Phillips DR. Molecular defects in interactions of platelets with the vessel wall. N Engl J Med 1984; 311: 1084-1098
    CrossrefPubMedGoogle Scholar
  • 2 Sakariassen KS, Nievelstein PFEM, Coller BS, Sixma JJ. The role of platelet membrane glycoproteins Ib and Ilb-IIIa in platelet adherence to human artery subendothelium. Br J Haematol 1986; 63: 681-691
    CrossrefPubMedGoogle Scholar
  • 3 Weiss HJ, Turitto VT, Baumgartner HR. Effect of sheat rate on platelet interaction with subendothelium in citrated and native blood. Shear-rate dependent decrease of adhesion in von-Willebrand's disease and the Bemard-Soulier syndrome. J Lab Clin Med 1978; 92: 750-764
    PubMedGoogle Scholar
  • 4 Weiss HJ, Hawiger J, Ruggeri ZM, Turitto VT, Thiagarajan P, Hoffmann T. Fibrinogen-independent platelet adhesion and thrombus formation on suben-dothelium mediated by glycoprotein Ilb/IIIa complex at high shear rate. J Clin Invest 1989; 83: 288-297
    CrossrefPubMedGoogle Scholar
  • 5 Ikeda Y, Handa M, Kawano K, Kamata T, Murata M, Araki Y, Anbo H, Kawai Y, Watanabe K, Itagaki I, Sakai K, Ruggeri ZM. The role of von Willebrand factor and fibrinogen in platelet aggregation under varying shear stress. J Clin Invest 1991; 87: 1234-1240
    CrossrefPubMedGoogle Scholar
  • 6 Girma JP, Meyer D, Verweij CL, Pannekoek H, Sixma JJ. Structure-function relationship of human von Willebrand factor. Blood 1987; 70: 605-611
    PubMedGoogle Scholar
  • 7 Fujimura YK, Titani K, Holland LZ, Roberts JR, Kostei P, Ruggeri ZM, Zimmerman TS. A heparin-binding domain of human von Willebrand factor. Characterization and localization to a tryptic fragment extending from amino acid residues Val-449 to Lys-728. J Biol Chem 1987; 262: 1734-1739
    PubMedGoogle Scholar
  • 8 Morton LF, Griffin B, Pepper DS, Barnes MJ. The interaction between collagens and factor VIII/von Willebrand factor: investigation of the structural requirements for interaction. Thromb Res 1983; 32: 545-556
    CrossrefPubMedGoogle Scholar
  • 9 Pareti FI, Niiya K, McPherson JM, Ruggeri ZM. Isolation and characterization of two domains of human von Willebrand factor that interact with fibrillar collagen types I and III. J Biol Chem 1987; 262: 13835-13841
    CrossrefPubMedGoogle Scholar
  • 10 Ruggeri ZM, De Macro, Gatti L, Bader R, Montgomery RR. Platelets have more than one binding site for von Willebrand factor. J Clin Invest 1983; 72: 1-12
    CrossrefPubMedGoogle Scholar
  • 11 Fujimura Y, Titani K, Holland LZ, Russel SR, Roberts JR, Eider JH, Ruggeri ZM, Zimmerman TS. A reduced and alkylated 52/48 kDa fragment beginning at amino acid residue 449 contains the domain interacting with platelet glycoprotein Ib. von-Willebrand factor. J Biol Chem 1986; 261: 381-385
    PubMedGoogle Scholar
  • 12 Mohri H, Fujimura Y, Shima M, Yoshioka A, Houghten RA, Ruggeri ZM, Zimmerman TS. Structure of the von Willebrand factor domain interacting with glycoprotein Ib. J Biol Chem 1988; 263: 17901-17904
    PubMedGoogle Scholar
  • 13 Mohri H, Yoshioka A, Zimmerman TS, Ruggeri ZM. Isolation of the von Willebrand factor domain interacting with platelet glycoprotein Ib, heparin and collagen and characterization of its three distinct functional sites. J Biol Chem 1989; 264: 17361-17367
    PubMedGoogle Scholar
  • 14 Azuma H, Dent JA, Sugimoto M, Ruggeri ZM, Ware J. Independent assembly and secretion of a dimeric adhesive domain of von Willebrand factor containing the glycoprotein Ib-bindig site. J Biol Chem 1991; 266: 12342-12347
    PubMedGoogle Scholar
  • 15 Sugimoto M, Ricca G, Hrinda ME, Schreiber AB, Searfoss GH, Bottini E, Ruggeri ZM. Functional modulation of the isolated glycoprotein Ib binding domain of von Willebrand factor expressed in Escherichia coli. Biochemistry 1991; 30: 5202-5209
    CrossrefPubMedGoogle Scholar
  • 16 Harker LA, Finch CA. Thrombokinetics in man. J Clin Invest 1969; 48: 963-974
    CrossrefPubMedGoogle Scholar
  • 17 Gospodarowicz D, Delgado D, Vlodavsky I. Permissive effect of the extracellular matrix on cell proliferation in vitro. Proc Natl Acad Sci USA 1980; 177: 4094-4098
    PubMedGoogle Scholar
  • 18 Lavee J, Martinowitz U, Mohr R, Goor D, Golan M, Langsam J, Malik Z, Savion N. The effect of transfusion of fresh whole blood versus platelet concentrates after cardiac operations. J Thorac Cardiovasc Surg 1989; 97: 204-212
    PubMedGoogle Scholar
  • 19 Vlodavsky I, Eldor A, HyAm E, Atzom R, Fuks Z. Platelet interaction with the extracellular matrix produced by cultured endothelial cells: a model to study the thrombogenicity of isolated subendothelial basal lamina. Thromb Res 1982; 28: 179-191
    CrossrefPubMedGoogle Scholar
  • 20 Baumgartner HR. The subendothelial surface and thrombosis. Thrombosis: Pathogenesis and clinical trials. Thromb Diath Haemorrh 1974; 59: 91 (suppl)
    PubMedGoogle Scholar
  • 21 Booyse FM, Feder S, Quarfoot AJ. Culture produced subendothelium. II. Effect of plasma, FVIIIR:WF and fibronectin on interaction of normal platelets with normal and von Willebrand porcine aortic subendothelium. Thromb Res 1982; 28: 299-311
    CrossrefPubMedGoogle Scholar
  • 22 Eldor A, Vlodavsky I, Martinowitz U, Fuks Z, Coller BS. Platelet interaction with subendothelial extracellular matrix: platelet-fibrinogen interactions are essential for platelet aggregation but not for the matrix-induced release reaction. Blood 1985; 65: 1477-1483
    PubMedGoogle Scholar
  • 23 Golan M, Modan M, Lavee J, Martinowitz U, Savion N, Goor DA, Mohr R. Transfusion of fresh whole blood stored (4° C) for short period fails to improve platelet aggregation on extracellular matrix and clinical hemostasis after cardiopulmonary bypass. J Thorac Cardiovasc Surg 1990; 99: 354-360
    PubMedGoogle Scholar
  • 24 Lavee J, Savion N, Smolinsky A, Goor DA. Platelet protection by aprotinin in cardiopulmonary bypass: electron microscopic study. Ann Thorac Surg 1992; 53: 477-481
    CrossrefPubMedGoogle Scholar
  • 25 Ruggeri ZM, Ware J. The structure and function of von Willebrand factor. Thromb Heamostas 1992; 67: 594-599
    PubMedGoogle Scholar
  • 26 Savage B, Shattil SJ, Ruggeri ZM. Modulation of platelet function through adhesion receptors: A dual role for glycoprotein IIb-IIIa (integrin αIIbβ3) mediated by fibrinogen and glycoprotein Ib-von Willebrand factor. J Biol Chem 1992; 267: 11300-11306
    PubMedGoogle Scholar
  • 27 Berliner S, Niiya K, Roberts JR, Houghten RA, Ruggeri ZM. Generation and characterization of peptide-specific antibodies that inhibit von Willebrand factor binding to glycoprotein IIb-IIIa without interacting with other adhesive molecules Selectivity is conferred by Pro and other amino acid residues adjacent to the sequence Arg1744-Gly1745-Asp1746 . J Biol Chem 1988; 263: 7500-7505
    PubMedGoogle Scholar
  • 28 De Marco, Girolami A, Zimmerman TS, Ruggeri ZM. von-Willebrand factor interaction with the glycoprotein IIb/IIIa complex: Its role in platelet function as demonstrated in patients with congenital afibrinogenemia. J Clin Invest 1986; 77: 1272-1277
    CrossrefPubMedGoogle Scholar
  • 29 Peterson DM, Stathopoulos NA, Giorgio TD, Heliums JD, Moake JL. Shear-induced platelet aggregation requires von Willebrand factor and platelet membrane glycoproteins Ib and IIb/IIIa. Blood 1987; 69: 625-628
    PubMedGoogle Scholar
  • 30 Pytela R, Pierschbacher MD, Ginsberg MH, Plow EF, Ruoslahti E. Platelet membrane glycoprotein Ilb/IIIa: Member of a family of Arg-Gly-Asp-specific adhesion receptors. Science 1986; 231: 1559-1562
    CrossrefPubMedGoogle Scholar
  • 31 De Marco, Girolami A, Russell S, Ruggeri ZM. Interaction of asialo von Willebrand factor with glycoprotein Ib induces fibrinogen binding to the glycoprotein Ilb/IIIa complex and mediates platelet aggregation. J Clin Invest 1985; 75: 1198-1203
    CrossrefPubMedGoogle Scholar
  • 32 Gralnick HR, Williams SB, Coller BS. Asialo von Willebrand factor interactions with platelets. Interdependence of glycoproteins Ib and IIb/IIIa for binding and aggregation. J Clin Invest 1985; 75: 19-25
    CrossrefPubMedGoogle Scholar
  • 33 Moake JL, Turner NA, Stathopoulos NA, Nolasco LH, Heliums JD. Involvement of large plasma von Willebrand factor (vWF) multimers and unusually large vWF forms derived from endothelial cells in shear-induced platelet aggregation. J Clin Invest 1986; 78: 1456-1461
    CrossrefPubMedGoogle Scholar