Download PDF
Thromb Haemost 1995; 73(03): 499-505
DOI: 10.1055/s-0038-1653804
Original Articles
Platelets
Schattauer GmbH Stuttgart

Glycoprotein IIb/IIIa Blockade Inhibits Platelet-mediated Force Development and Reduces Gel Elastic Modulus

Marcus E Carr Jr
1   The Coagulation Special Studies Laboratory, Departments of Medicine and Pathology, Medical College of Virginia, USA
2   McGuire V.A. Medical Center, Richmond, VA, USA
,
Sheryl L Carr
1   The Coagulation Special Studies Laboratory, Departments of Medicine and Pathology, Medical College of Virginia, USA
,
Roy R Hantgan
3   Bowman Gray School of Medicine, Winston-Salem, NC, USA
,
Julie Braaten
3   Bowman Gray School of Medicine, Winston-Salem, NC, USA
› Author Affiliations
Further Information

Publication History

Received 18 January 1994

Accepted after resubmission 08 November 1994

Publication Date:
09 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

The effects of GPIIb/IIIa blockade on clot retraction were studied utilizing an instrument which directly measures force produced by platelets. GPIIb/IIIa disruption by calcium chelation, and GPIIb/IIIa blockade by peptides and anti-GPIIb/IIIa antibodies were investigated. One mM EDTA suppressed ADP-induced platelet aggregation by 72% and reduced force developed at 1200 s by 33%. At 234 μM, the tetrapeptide Arg-Gly-Asp-Ser (RGDS) suppressed platelet aggregation by 74%, reduced force at 1200 s by 45% and reduced gel elastic modulus by 19%. At 10 μM, the peptide D-Arg-Gly-L-Asp-L-Try (D-RGDW) completely suppressed platelet aggregation, reduced force development by 38% and reduced gel elastic modulus by 29%. At 0.133 μM, monoclonal anti-GPIIIa antibody (AP-3) reduced force development by 74% and reduced gel modulus by 60%. Murine antiGPIIb/IIIa antibodies 10E5 and 7E3 markedly suppressed force development. At 0.133 μM, 10E5 reduced force by 89% and reduced gel modulus by 67%. At 0.053 μM, 7E3 completely stopped force development and reduced gel modulus by 46%. Platelet aggregation was blocked by 0.027 μM 7E3. Selective GPIIb blockade by antibodies did not affect force development. None of the agents studied altered fibrin structure as monitored by effects on fibrin mass/length ratios. Suppression of platelet aggregation occurred at inhibitor concentrations substantially lower than those required to suppress force development. Complete suppression of platelet aggregation did not assure inhibition of clot retraction probably due to profound platelet activation by thrombin. These results indicate that inhibition of fibrin(ogen) binding to GPIIb/IIIa, either by disruption of GPIIb/IIIa or by competitive blockade, inhibits platelet mediated force development and results in clot structures which are substantially less resistant to deformation by outside forces.

 

  • References

  • 1 Budtz-Olsen OE. Clot Retraction. Blackwell, Oxford; England: 1951
    Google Scholar
  • 2 Morgenstern E, Korell U, Richter J. Platelets and fibrin strands during clot retraction. Thromb Res 1984; 33: 617-623
    CrossrefPubMedGoogle Scholar
  • 3 Cohen I, Gerrard JM, White JG. Ultrastructure of clots during isometric contraction. J Cell Biol 1982; 93: 775-787
    CrossrefPubMedGoogle Scholar
  • 4 Chao FC, Shepro D, Tullis JL, Belamarich FA, Curby WA. Similarities between platelet contraction and cellular motility during mitosis: Role of platelet microtubules in clot retraction. J Cell Sci 1976; 20: 569-588
    PubMedGoogle Scholar
  • 5 Carr ME, Zekert SL. Measurement of platelet-mediated force development during plasma clot formation. Am J Med Sci 1991; 302: 13-18
    CrossrefPubMedGoogle Scholar
  • 6 Carr ME, Zekert SL. Force monitoring of clot retraction during DDAVP therapy for the qualitative platelet disorder of uraemia: report of a case. Blood Coagul Fibrinolysis 1991; 2: 303-308
    CrossrefPubMedGoogle Scholar
  • 7 Greilich P, Carr M, Zekert S. Effects of aspirin on clot structure and fibrin platelet interactions in patients with severe coronary artery disease. Clin Res 1992; 40: 241 A
    PubMedGoogle Scholar
  • 8 Gartner TK, Ogilvie ML. Peptides and monoclonal antibodies which bind to platelet glycoproteins IIb and/or IIIa inhibit clot retraction. Thromb Res 1988; 49: 43-53
    CrossrefPubMedGoogle Scholar
  • 9 Hantgan RR. Localization of the domains of fibrin involved in binding to platelets. Biochim Biophys Acta 1988; 968: 36-44
    CrossrefPubMedGoogle Scholar
  • 10 Zucher MB, Grant RA. Nonreversible loss of platelet aggregability induced by calcium deprivation. Blood 1978; 52: 505-514
    PubMedGoogle Scholar
  • 11 Fitzgerald IA, Phillips DR. Calcium regulation of the platelet membrane glycoprotein IIb-IIIa complex. J Biol Chem 1985; 260: 11366-11374
    PubMedGoogle Scholar
  • 12 Plow EF, Pierschbacher MD, Ruoslahti E, Margueric GA, Ginsberg MH. Arginyl-glycyl-aspartic acid sequences and fibrinogen binding to platelets. Blood 1985; 70: 110-115
    PubMedGoogle Scholar
  • 13 Hantgan RR, Eudenburg SC, Cavero I, Marguerie G, Uzan A, Sixma JJ, de Groot PG. Inhibition of platelet adhesion to fibrin(ogen) in flowing whole blood by Arg-Gly-Asp and fibrinogen γ-chain carboxy terminal peptides. Thromb Haemost 1992; 68: 694-700
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Newman PJ, Allen RW, Kahn RA, Kunicki TJ. Quantitation of membrane glycoprotein IIIa on intact human platelets using the monoclonal antibody, AP-3. Blood 1985; 65: 227-232
    PubMedGoogle Scholar
  • 15 Coller BS. A new murine monoclonal antibody reports an activationdepen- dent change in the conformation and/or microenvironment of the platelet glycoprotein IIb/IIIa complex. J Clin Invest 1985; 76: 101-108
    CrossrefPubMedGoogle Scholar
  • 16 Coller BS, Peerschke El, Seligsohn U, Scudder LE, Nurden AT, Rosa JP. Studies on the binding of an alloimmune and two murine monoclonal antibodies to the platelet glycoprotein IIb/IIIa complex receptor. J Lab Clin Med 1986; 107: 384-392
    PubMedGoogle Scholar
  • 17 Clauss A. Rapid physiological coagulation method in determination of fibrinogen. Acta Haematol 1957; 17: 237-246
    CrossrefPubMedGoogle Scholar
  • 18 Coller BS, Peerschke El, Scudder LE, Sullivan CA. A murine monoclonal 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: 325-338
    CrossrefPubMedGoogle Scholar
  • 19 Carr ME, Gabriel DA. The effect of dextran 70 on the structure of plasma- derived fibrin gels. J Lab Clin Med 1980; 96: 985-993
    PubMedGoogle Scholar
  • 20 Carr ME, Gabriel DA. Dextran-induced changes in fibrin fiber size and density based on wavelength dependence of gel turbidity. Macromolecules 1980; 13: 1473-1477
    CrossrefPubMedGoogle Scholar
  • 21 Carr ME, Hermans J. Size and density of fibrin fibers from turbidity. Macromolecules 1978; 11: 46-50
    CrossrefPubMedGoogle Scholar
  • 22 Corn M, Jackson DP, Conley CL. Components of blood necessary for clot retraction. Bull Johns Hopkins Hosp 1960; 107: 90-104
    PubMedGoogle Scholar
  • 23 Fujimura K, Phillips DR. Calcium cation regulation of glycoprotein IIb/IIIa complex formation in platelet plasma membrane. J Biol Chem 1983; 258: 10247-10252
    PubMedGoogle Scholar
  • 24 Wencel-Drake JD, Plow EF, Kunicki TJ, Woods VL, Keller DM, Ginsberg MH. Localization of internal pools of membrane glycoproteins involved in platelet adhesive responses. Am J Pathol 1986; 124: 324-334
    PubMedGoogle Scholar
  • 25 Gogstad GD, Hagen I, Korsmo R, Solum NO. Characterization of the proteins of isolated human platelet alpha-granules. Evidence for a separate alpha-granule-pool of the glycoproteins IIb and IIIa Biochim Biophys Acta 1981; 670: 150-162
    CrossrefPubMedGoogle Scholar
  • 26 Shattil SJ, Hoxie JA, Cunningham M, Brass LF. Changes in the platelet membrane glycoprotein Ilb. IIIa complex during platelet activation J Biol Chem 1985; 260: 11107-11114
    PubMedGoogle Scholar
  • 27 Hantgan RR, Nichols WL, Ruggeri ZM. von Willebrand factor competes with fibrin for occupancy of GPIIb:IIa on thrombin-stimulated platelets. Blood 1990; 75: 889-894
    PubMedGoogle Scholar
  • 28 Cohen I, Burk DL, White JG. The effect of peptides and monoclonal antibodies that bind to platelet glycoprotein IIb-IIIa complex on the development of clot tension. Blood 1989; 73: 1880-1887
    PubMedGoogle Scholar
  • 29 Thomas DP. The role of platelet catecholamines in the aggregation of platelets by collagen and thrombin. Exp Biol Med 1968; 3: 129-134
    PubMedGoogle Scholar
  • 30 Peerschke EI B. Effect of epinephrine on fibrinogen receptor exposure by aspirin-treated platelets and platelets from concentrates in response to ADP and thrombin. Am J Hematol 1984; 16: 335-345
    CrossrefPubMedGoogle Scholar
  • 31 Plow EF, Marguerie GA. Participation of ADP in the binding of fibrinogen to thrombin-stimulated platelets. Blood 1980; 56: 553-555
    PubMedGoogle Scholar
  • 32 Bettelheim FR, Bailey K. The products of the action of thrombin on fibrinogen. Biochim Biophys Acta 1952; 9: 578-579
    CrossrefPubMedGoogle Scholar
  • 33 Lorand L. Fibrinopeptide. Biochem J 1952; 52: 200-203
    CrossrefPubMedGoogle Scholar
  • 34 Hantgan RR, Hermans J. Assembly of fibrin. A light scattering study J Biol Chem 1979; 254: 11272-11281
    PubMedGoogle Scholar
  • 35 Haverstick DM, Cowan JF, Yamada KM, Santoro SA. Inhibition of platelet adhesion to fibronectin, fibrinogen, and von Willebrand factor substrates by a synthetic tetrapeptide derived from the cell-binding domain of fibronectin. Blood 1985; 66: 946-952
    PubMedGoogle Scholar
  • 36 Lewis JC, Hantgan RR, Stevenson SC, Thornburg T, Kieffer N, Guichard J, Breton-Gorius J. Fibrinogen and glycoprotein IIb/IIIa localization during platelet adhesion. Localization to the granulomere and at sites of platelet interaction Am J Pathol 1990; 136: 239-252
    PubMedGoogle Scholar
  • 37 Lindon JN, McManama G, Kushner L, Merril EW, Salzman EW. Does the conformation of adsorbed fibrinogen dictate platelet interactions with artificial surfaces. Blood 1986; 68: 355-362
    PubMedGoogle Scholar
  • 38 Savage B, Ruggeri ZM. Selective recognition of adhesive sites in surface- bound fibrinogen by glycoprotein IIb-IIIa on nonactivated platelets. J Biol Chem 1991; 266: 11227-11233
    PubMedGoogle Scholar
  • 39 Pierschbacher MD, Ruoslahti E. Influence of stereochemistry of the sequence arg-gly-asp-Xaa on binding specificity in cell adhesion. J Biol Chem 1987; 262: 17294-17298
    PubMedGoogle Scholar
  • 40 Tranqui L, Andrieux A, Hudry-Clergeon G, Ryckevaert JJ, Soyez S, Chapel A, Ginsberg MH, Plow EF, Marguerie G. Differential structural requirements for fibrinogen binding to platelets and to endothelial cells. J Cell Biol 1989; 108: 2519-2527
    CrossrefPubMedGoogle Scholar
  • 41 Newman PJ, McEver RP, Doers MP, Kunicki TJ. Synergistic action of two murine monoclonal antibodies that inhibit ADP-induced platelet aggregation without blocking fibrinogen binding. Blood 1987; 69: 668-676
    PubMedGoogle Scholar
  • 42 Jennings LK, Fitzgerald D, White MM. An active conformation of platelet GPIIb/IIIa that binds to fibrin(ogen) does not mediate clot retraction. Blood 1993; 82: 211a
    PubMedGoogle Scholar
  • 43 Coller BS, Folts JD, Smith SR, Scudder LE, Jordan R. Abolition of in vitro platelet thrombus formation in primates with monoclonal antibodies to the platelet GPIIb/IIIa receptor - Correlation with bleeding time, platelet aggregation, and blockade of GPIIb/IIIa receptors. Circulation 1989; 80: 174-176
    PubMedGoogle Scholar
  • 44 Gold HK, Gimple LW, Yasuda T, Leinbach RC, Werner W, Holt R, Jordan R, Berger H, Collen D, Coller BS. Pharmacodynamic study of F(ab')2 fragments of murine monoclonal antibody 7E3 directed against human platelet glycoprotein IIb/IIIa in patients with unstable angina pectoris. J Clin Invest 1990; 86: 651-659
    CrossrefPubMedGoogle Scholar
  • 45 Jen CJ, McIntire LV. The structural properties and contractile force of a clot. Cell Motil 1982; 2: 445-455
    PubMedGoogle Scholar