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Thromb Haemost 1998; 80(03): 428-436
DOI: 10.1055/s-0037-1615225
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Ristocetin- and Thrombin-induced Platelet Aggregation at Physiological Shear Rates: Differential Roles for GPIb and GPIIb-IIIa Receptor[ 1 ]

A. Kasirer-Friede
1   From the Depts. of Physiology and Experimental Medicine, McGill University, Montreal, Que., Canada
,
M. M. Frojmovic
1   From the Depts. of Physiology and Experimental Medicine, McGill University, Montreal, Que., Canada
› Author Affiliations
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Publication History

Received 05 November 1997

Accepted after resubmission 15 May 1998

Publication Date:
08 December 2017 (online)

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Summary

We recently reported that washed platelets (WP) activated with ADP and expressing surface-bound vWF aggregated in flow through small tubes or in a cylindrical couette device at physiological shear rates of G = 300 s–1-1000 s–1 in the absence of exogenous ligands, with GPIb-vWF partially, and activated GPIIb-IIIa totally required for the aggregation. We have now extended these studies to aggregation of platelets “activated” with ristocetin or thrombin. Washed platelet suspensions with added soluble vWF and ristocetin (0.3-0.75 mg/ml), or activated with thrombin (0.01-0.5 U/ml) but no added ligand, were sheared in a coaxial cylinder device at uniform shear rate, G = 1000 s–1. The collision capture efficiency (αG) with which small aggregates form (= experimental/calculated initial rates of aggregation) was correlated with vWF platelet binding assessed by flow cytometry. The vWF-GPIb interaction was exclusively able to support ristocetin-mediated shear aggregation of metabolically active platelets, with very few vWF monomer equivalents bound per platelet (representing ≤10 molecules of 10 million Da) required to yield high capture efficiencies (αG = 0.38 ± .02; n = 11), suggesting rapid and stable bond formations between vWF and GPIb. However, platelet surface-expressed vWF, generated by addition of thrombin to washed platelets, was found to mediate platelet aggregation with αG = 0.08 ± .01 (n = 6), surprisingly comparable to that previously reported for WP and ADP activation. Blocking the GPIIb-IIIa receptor decreased αG by 95 ± 3% (n = 3), while a monoclonal antibody to the vWF site on GPIb caused a 49 ± 7% (n = 8) decrease in αG. The partial role for GPIb thus appears to reflect a facilitative function for increasing contact time between flowing platelets, and allowing engagement of the GPIIb-IIIa receptor to yield stable attachment.

1 We acknowledge the Medical Research Council of Canada for research and personnel support.


 

  • References

  • 1 Savage B, Salvidar E, Ruggeri ZM. Initiation of platelet adhesion by arrest onto fibrinogen or translocation on von Willebrand factor. Cell 1996; 84: 289-97.
    CrossrefPubMedGoogle Scholar
  • 2 Lankhof H, Wu YP, Vink T, Schiphorst ME, Zerwes HG, de Groot P, Sixma JJ. Role of the glycoprotein Ib-binding A1 repeat and the RGD sequence in platelet adhesion to human recombinant von Willebrand factor. Blood 1995; 86: 1035-42.
    PubMedGoogle Scholar
  • 3 Ikeda Y, Handa M, Kamata T, Kawano K, Kawai Y, Watanabe K, Kawakami K, Sakai K, Fukuyama M, Itagaki I. et al. Transmembrane calcium influx associated with von Willebrand factor binding to GPIb in the initiation of shear-induced platelet aggregation. Thromb Haemost 1993; 69: 496-502.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Moake JL, Turner NA, Stathopoulos NA, Nolasco LH, Hellums JD. Shear-induced platelet aggregation can be mediated by vWF released from platelets, as well as by exogenous large or unusually large vWF multimers, requires adenosine diphosphate and is resistant to aspirin. Blood 1988; 71: 1366-74.
    PubMedGoogle Scholar
  • 5 Fujimoto T, Hawiger J. Adenosine diphosphate induces binding of von Willebrand factor to human platelets. Nature 1982; 297: 154-6.
    CrossrefPubMedGoogle Scholar
  • 6 Gralnick HR, Williams SB, Coller B. Fibrinogen competes with von Willebrand factor for binding to the Glycoprotein IIb/IIIa complex when platelets are stimulated with thrombin. Blood 1984; 64: 797-800.
    PubMedGoogle Scholar
  • 7 Scott JP, Montgomery RR, Retzinger GS. Dimeric ristocetin flocculates proteins, binds to platelets, and mediates von-Willebrand factor-dependent agglutination of platelets. J Biol Chem 1991; 266: 8149-55.
    PubMedGoogle Scholar
  • 8 Goto S, Salomon DR, Ikeda Y, Ruggeri ZM. Characterization of the unique mechanism mediating the shear-dependent binding of soluble von Willebrand factor to platelets. J Biol Chem 1995; 270: 23352-61.
    CrossrefPubMedGoogle Scholar
  • 9 Ruggeri ZM, De Marco L, 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
  • 10 Taylor AD, Neelamegham S, Hellums JD, Smith CW, Simon SI. Molecular dynamics of the transition from L-selectin to β2-integrin dependent neutrophil adhesion under defined hydrodynamic shear. Biophys J 1996; 71: 3488-500.
    CrossrefPubMedGoogle Scholar
  • 11 Frojmovic MM, Kasirer-Friede A, Goldsmith HL, Brown EA. Surface-secreted von Willebrand factor mediates aggregation of ADP-activated platelets at moderate shear stress: facilitated by GPIb but controlled by GPIIb-IIIa. Thromb Haemost 1997; 77: 568-76.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 12 Goldsmith HL, Frojmovic MM, Braovac S, McIntosh F, Wong T. Adenosine diphosphate-induced aggregation of human platelets in flow through tubes. III. Shear and extrinsic fibrinogen-dependent effects. Thromb Haemost 1994; 71: 78-90.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Goto S, Ikeda Y, Salvidar E, Ruggeri ZM. Distinct mechanisms of platelet aggregation as a consequence of different shearing flow conditions. J Clin Invest 1998; 101: 479-86.
    CrossrefPubMedGoogle Scholar
  • 14 Plow EF, Pierschbacher MD, Ruoslahti E, Marguerie GA, Ginsberg MH. The effect of Arg-Gly-Asp-containing peptides on fibrinogen and von Willebrand factor binding to platelets. Proc Natl Acad Sci USA 1985; 82: 8057-61.
    CrossrefPubMedGoogle Scholar
  • 15 Kouns WC, Kirchofer D, Hadváry P, Edenhofer A, Weller T, Pfenninger G, Baumgartner HR, Jennings LK, Steiner B. Reversible conformational changes induced in Glycoprotein GP IIb-IIIa by a potent and selective peptidomimetic inhibitor. Blood 1992; 10: 2539-47.
    PubMedGoogle Scholar
  • 16 Schor K, Darius H, Matzky R, Ohlendort R. The antiplatelet and cardiovascular actions of a new carbaxylic derivative (ZK 36374)-equipotent to PGI2 in vitro. Arch Pharm 1981; 316: 252-5.
    CrossrefPubMedGoogle Scholar
  • 17 Coller BS, Peerschke EI, Scudder LE, Sullivan CA. Studies with a murine monoclonal antibody that abolishes ristocetin-induced binding of von Willebrand factor to platelets; additional evidence in support of GPIb as a platelet receptor for von Willebrand factor. Blood 1983; 61: 99-110.
    PubMedGoogle Scholar
  • 18 Coller BS, Peerschke EI, Scudder LE, Sullivan CA. A murine monoclonal antibody that completely blocks the binding of fibrinogen to platelets produces a thrombasthenic-like state in normal platelets and binds to glycoproteins IIb and/or IIIa. J Clin Invest 1983; 72: 325-38.
    CrossrefPubMedGoogle Scholar
  • 19 Shattil SJ, Cunningham M, Hoaxie JA. Detection of activation platelets in whole blood using activation-dependent monoclonal antibodies and flow cytometry. Blood 1987; 70: 307-15.
    PubMedGoogle Scholar
  • 20 Handa M, Titani K, Holland LZ, Roberts JR, Ruggeri ZM. The von Willebrand factor-binding domain of platelet membrane glycoprotein GPIb. J Biol Chem 1986; 261: 12579-85.
    PubMedGoogle Scholar
  • 21 Weinstein M, Vosburgh E, Phillips M, Turner N, Chute-Rose L, Moake J. Isolation from commercial aurin tricarboxylic acid of the most effective polymeric inhibitors of von Willebrand factor interaction with glycoprotein Ib. Comparison with other polyanionic and polyaromatic polymers. Blood 1991; 78: 2291-8.
    PubMedGoogle Scholar
  • 22 Girma JP, Fressinaud E, Christophe O, Roualt C, Obert B, Takahashi Y, Meyer D. Aurin tricarboxylic acid inhibits platelet adhesion to collagen by binding to the 509-695 disulphide loop of von Willebrand factor and competing with glycoprotein Ib. Thromb Haemost 1992; 68: 707-13.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Ruggeri ZM, Zimmerman TS, Russell S, Bader R, deMarco L. Purification of von Willebrand factor. Methods in Enzymology 1992; 215: 265-8.
    PubMedGoogle Scholar
  • 24 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII/von Willebrand factor. Blood 1981; 57: 1140-3.
    PubMedGoogle Scholar
  • 25 Zaleski A, Henriksen RA. Visualization of the multimeric structure of von Willebrand factor using a peroxidase-conjugated second antibody. J Lab Clin Med 1986; 107: 172-5.
    PubMedGoogle Scholar
  • 26 Tang SS, Frojmovic MM. The effects of pCO2 and pH on platelet shape change and aggregation for human and rabbit platelet-rich plasma. Thromb Res 1977; 10: 135-45.
    CrossrefPubMedGoogle Scholar
  • 27 The TH, Feltkamp TEW. Conjugation of fluorescein isothiocyanate to antibodies. I. Experiments on the conditions of conjugation. Immunol 1970; 18: 865-73.
    PubMedGoogle Scholar
  • 28 Xia Z, Wong T, Liu Q, Kasirer-Friede A, Brown E, Frojmovic MM. Optimally functional fluorescein isothiocyanate-labelled fibrinogen for quantitative studies of binding to activated platelets and platelet aggregation. Br J Haematol 1996; 93: 204-14.
    CrossrefPubMedGoogle Scholar
  • 29 Williams SB, McKeown LP, Krutzch H, Hansmann Gralnick H. Purification and characterization of human platelet von Willebrand factor. Br J Haematol 1994; 88: 582-91.
    CrossrefPubMedGoogle Scholar
  • 30 Frojmovic MM, Mooney RF, Wong T. Dynamics of platelet glycoprotein IIb-IIIa receptor expression and fibrinogen binding. I. Quantal activation of platelet subpopulations varies with adenosine diphosphate concentration. Biophys J 1994; 67: 2060-8.
    CrossrefPubMedGoogle Scholar
  • 31 Xia A, Frojmovic MM. Aggregation efficiency of activated normal or fixed platelets in a simple shear field: Effect of shear and fibrinogen occupancy. Biophys J 1994; 66: 2190-201.
    CrossrefPubMedGoogle Scholar
  • 32 Smoluchowski M. von. Versuch einer mathematischen Theorie der Koagulationskinetik kolloider Lösungen. Z Phys Chem 1917; 92: 129-68.
    PubMedGoogle Scholar
  • 33 Harrison RL, McKee PA. Comparison of thrombin and ristocetin in the interaction between von Willebrand factor and platelets. Blood 1983; 62: 346-53.
    PubMedGoogle Scholar
  • 34 Azzam K, Cissé-Thiam M, Drouet L. The antithrombotic effect of aurin tricarboxylic acid in the guinea pig is not solely due to the interaction with the von Willebrand factor-GPIb axis. Thromb Haemost 1996; 75: 203-10.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Greco NJ, Jones GD, Tandon NN, Kornhauser R, Jackson B, Jamieson GA. Differentiation of the two forms of GPIb functioning as receptors for alpha-thrombin and von Willebrand factor: Ca2+ responses of protease-treated human platelets activated with alpha thrombin and the tethered ligand peptide. Biochem 1996; 35: 915-21.
    CrossrefPubMedGoogle Scholar
  • 36 Siedlicki CA, Lestini BJ, Kottke-Marchant K, Eppell SJ, Wilson DL, Marchant RE. Shear-dependent changes in the three-dimensional structure of human von Willebrand factor. Blood 1996; 88: 2939-50.
    PubMedGoogle Scholar
  • 37 Lawrence MB, Springer TA. Leukocytes roll on a selectin at physiologic flow rates: distinction from and prerequisite for adhesion to integrins. Cell 1991; 65: 859-73.
    CrossrefPubMedGoogle Scholar
  • 38 Ikeda Y, Handa M, Kawano K, Kamata T, Murata M, Araki Y, Anbo H, Kawai Y, Watanabe K, Itagaki I, Sakai K, Ruggeri Z. The role of von Willebrand factor and fibrinogen in platelet aggregation under varying shear stress. J Clin Invest 1991; 87: 1234-40.
    CrossrefPubMedGoogle Scholar
  • 39 Ozaki Y, Satoh K, Yatomi Y, Miura S, Fujimura Y, Kume S. Protein tyrosine phosphorylation in human platelets induced by interaction between glycoprotein Ib and von Willebrand factor. Biochim Biophys Act 1995; 1243: 482-8.
    CrossrefPubMedGoogle Scholar
  • 40 Chediak J, Telfer C, Laan BV, Maxey B, Cohen I. Cycles of agglutination-disagglutination induced by ristocetin in thrombasthenic platelets. Br J Haematol 1979; 43: 113-26.
    CrossrefPubMedGoogle Scholar
  • 41 Manley RStJ, Mason SG. Particle motions in sheared suspensions. II. Collisions of uniform spheres. J Colloid Sci 1952; 7: 354-69.
    CrossrefPubMedGoogle Scholar
  • 42 Greco NJ, Tenner Jr. TE, Tandon NN, Jamieson GA. PPACK-Thrombin inhibits thrombin-induced platelet aggregation and cytoplasmic acidification, but does not inhibit platelet shape change. Blood 1990; 75: 1983-90.
    PubMedGoogle Scholar
  • 43 Bell DN, Spain S, Goldsmith HL. The ADP-induced aggregation of human platelets in flow through tubes: I. Measurement of the concentration and size of single platelets and aggregates. Biophys J 1989; 56: 817-28.
    CrossrefPubMedGoogle Scholar
  • 44 Bell DN, Spain S, Goldsmith HL. The ADP-induced aggregation of human platelets in flow through tubes: II. Effect of shear rate, donor sex, and ADP concentration. Biophys J 1989; 56: 829-43.
    CrossrefPubMedGoogle Scholar
  • 45 Frojmovic MM, Mooney RF, Wong T. Dynamics of platelet glycoprotein IIb-IIIa expression and fibrinogen binding. II. Quantal activation parallels platelet capture in stir-associated microaggregation. Biophys J 1994; 67: 2069-75.
    CrossrefPubMedGoogle Scholar
  • 46 Suzuki H, Kinlough-Rathbone RL, Packham MA, Tanoue K, Yamazaki H, Fraser M. Immunocytochemical localization of fibrinogen during thrombin-induced aggregation of washed human platelets. Blood 1988; 71: 1310-20.
    PubMedGoogle Scholar
  • 47 Heilmann E, Hourdille P, Provost A, Papponneau A, Nurden AT. Thrombin-induced platelet aggregates have a dynamic structure. Arterioscl and Thrombos 1991; 11: 704-18.
    PubMedGoogle Scholar
  • 48 Dong JF, Hyun W, Lopez JA. Aggregation of mammalian cells expressing the platelet glycoprotein (GP) Ib-IX complex and the requirement for tyrosine sulfation of GP Ib alpha. Blood 1995; 86: 4175-83.
    PubMedGoogle Scholar
  • 49 Berger G, Massé JM, Cramer EM. Alpha-granule membrane mirrors the platelet plasma membrane and contains the glycoproteins Ib, IX and V. Blood 1996; 87: 1385-95.
    PubMedGoogle Scholar
  • 50 Parker RI, Shafer BC, Gralnick HR. Platelet density-dependent partition of platelet von Willebrand factor between alpha granule and non-alpha granule pools. Thromb Haemost 1987; 58: 911-4.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 51 Parker RI, Rick ME, Gralnick HR. Effect of calcium on the availability of platelet von Willebrand factor. J Lab Clin Med 1985; 106: 336-42.
    PubMedGoogle Scholar
  • 52 Harrison P, Cramer EM. Platelet α-granules. Blood Reviews 1993; 7: 52-62.
    CrossrefPubMedGoogle Scholar