Thromb Haemost 2001; 85(06): 1090-1096
DOI: 10.1055/s-0037-1615969
Review Article
Schattauer GmbH

A New Approach in the Study of the Molecular and Cellular Events Implicated in Heparin-induced Thrombocytopenia

Formation of Leukocyte-platelet Aggregates
Mahnouch Khairy
1   INSERM-U428, Faculté de Pharmacie (Université Paris V), Paris, France
,
Dominique Lasne
1   INSERM-U428, Faculté de Pharmacie (Université Paris V), Paris, France
,
Brigitte Brohard-Bohn
1   INSERM-U428, Faculté de Pharmacie (Université Paris V), Paris, France
,
Martine Aiach
1   INSERM-U428, Faculté de Pharmacie (Université Paris V), Paris, France
,
Francine Rendu
1   INSERM-U428, Faculté de Pharmacie (Université Paris V), Paris, France
,
Christilla Bachelot-Loza
1   INSERM-U428, Faculté de Pharmacie (Université Paris V), Paris, France
› Author Affiliations
Further Information

Publication History

Received 09 November 2000

Accepted after revision 25 January 2001

Publication Date:
12 December 2017 (online)

Summary

Heparin-induced thrombocytopenia (HIT), a relatively common complication of heparin therapy, results of platelet activation, via the receptor for the Fc domain of IgG (FcγRIIa), by heparin-dependentantibodies, commonly directed against the heparin-platelet factor 4 (H-PF4) antigenic complex. Our strategy was to use whole blood allowing the study of leukocyte-platelet interactions. Experiments were performed with blood from healthy donors incubated with HIT patients’ plasma and different concentrations of heparin. We showed that 75% of the HIT patients’ plasma induced the formation of leukocyteplatelet-aggregates in a heparin-dependent-manner. The formation of leukocyteplatelet-aggregates induced by HIT plasma in the presence of heparin was (i) independent of the healthy blood donor FcγRIIa polymorphism, (ii) correlated with the levels of anti H-PF4 IgG antibodies contained in the patients’ plasma, and to a lesser extent to anti H-PF4 IgM antibodies, and (iii) was mediated by P-selectin. This report opens new prospects in the study of the molecular and cellular events implicated in HIT.

 
  • References

  • 1 Chong BH. Heparin-induced thrombocytopenia. Br J Haematol 1995; 89: 431-9.
  • 2 Warkentin TE, Chong BH, Greinacher A. Heparin-induced thrombocytopenia: towards consensus. Thromb Haemost 1998; 79: 1-7.
  • 3 Warkentin TE. Heparin-induced thrombocytopenia: a clinicopathologic syndrome. Thromb Haemost 1999; 82: 439-47.
  • 4 Chong BH, Pitney WR, Castaldi PA. Heparin-induced thrombocytopenia: association of thrombotic complications with heparin-dependent IgG antibody that induces thromboxane synthesis in platelet aggregation. Lancet 1982; 4: 1246-9.
  • 5 Amiral J, Bridey F, Dreyfus M, Vissoc AM, Fressinaud E, Wolf M, Meyer D. Platelet factor 4 complexed to heparin is the target for antibodies generated in heparin-induced thrombocytopenia. Thromb Haemost 1992; 68: 95-6.
  • 6 Greinacher A, Potzsch B, Amiral J, Dummel V, Eichner A, Mueller-Eckhardt C. Heparin-associated thrombocytopenia: isolation of the antibody and characterization of a multimolecular PF4-heparin complex as the major antigen. Thromb Haemost 1994; 71: 247-51.
  • 7 Kelton JG, Smith JW, Warkentin TE, Hayward CP, Denomme GA, Horsewood P. Immunoglobulin G, from patients with heparin-induced thrombocytopenia binds to a complex of heparin and platelet factor 4. Blood 1994; 83: 3232-9.
  • 8 Visentin GP, Ford SE, Scott JP, Aster RH. Antibodies from patients with heparin-induced thrombocytopenia/thrombosis are specific for platelet factor 4 complexed with heparin or bound to endothelial cells. J Clin Invest 1994; 93: 81-8.
  • 9 Amiral J, Bridey F, Wolf M, Boyer-Neumann C, Fressinaud E, Vissac AM, Peynaud-Debayle E, Dreyfus M, Meyer D. Antibodies to macromolecular platelet factor 4-heparin complexes in heparin-induced thrombocytopenia: a study of 44 cases. Thromb Haemost 1995; 73: 21-8.
  • 10 Greinacher A, Alban S, Dummel V, Franz G, Mueller-Eckhardt C. Characterization of the structural requirements for a carbohydrate based anticoagulant with a reduced risk of inducing the immunological type of heparin-associated thrombocytopenia. Thromb Haemost 1995; 74: 886-92.
  • 11 Horsewood P, Warkentin TE, Hayward CP, Kelton JG. The epitope specificity of heparin-induced thrombocytopenia. Br J Haematol 1996; 95: 161-7.
  • 12 Kelton JG, Sheridan D, Santos A, Smith J, Steeves K, Smith C, Brown C, Murphy WG. Heparin-induced thrombocytopenia: laboratory studies. Blood 1988; 72: 925-30.
  • 13 Chong BH, Fawaz I, Chesterman CN, Berndt MC. Heparin-induced thrombocytopenia: mechanism of interaction of the heparin-dependent antibody with platelets. Br J Haematol 1989; 73: 235-40.
  • 14 Newman PM, Chong B. Heparin-induced thrombocytopenia: new evidence for the dynamic binding of purified anti-PF4-heparin antibodies to platelets and the resultant platelet activation. Blood 2000; 96: 182-7.
  • 15 Amiral J, Wolf M, Fischer A, Boyer-Neumann C, Vissac A, Meyer D. Pathogenicity of IgA and/or IgM antibodies to heparin-PF4 complexes in patients with heparin-induced thrombocytopenia. Br J Haematol 1996; 92: 954-9.
  • 16 Suh JS, Malik MI, Aster RH, Visentin GP. Characterization of the humoral immune response in heparin-induced thrombocytopenia. Am J Hematol 1997; 54: 196-201.
  • 17 Greinacher A, Amiral J, Dummel V, Vissac A, Kiefel V, Mueller-Eckhardt C. Laboratory diagnosis of heparin-associated thrombocytopenia and comparison of platelet aggregation test, heparin-induced platelet activation test, and platelet factor 4/heparin enzyme-linked immunosorbent assay. Transfusion 1994; 34: 381-5.
  • 18 Bachelot-Loza C, Saffroy R, Lasne D, Chatellier G, Aiach M, Rendu F. Importance of the FcgammaRIIa-Arg/His-131 polymorphism in heparin-induced thrombocytopenia diagnosis. Thromb Haemost 1998; 79: 523-8.
  • 19 Warkentin TE, Levine MN, Hirsh J, Horsewood P, Roberts RS, Gent M, Kelton JG. Heparin-induced thrombocytopenia in patients treated with low-molecular-weight heparin or unfractionated heparin. N Engl J Med 1995; 332: 1330-5.
  • 20 Amiral J, Peynaud-Debayle E, Wolf M, Bridey F, Vissac AM, Meyer D. Generation of antibodies to heparin-PF4 complexes without thrombocytopenia in patients treated with unfractionated or low-molecular-weight heparin. Am J Hematol 1996; 52: 90-5.
  • 21 Warkentin TE, Sheppard JA, Horsewood P, Simpson PJ, Moore JC, Kelton JG. Impact of the patient population on the risk for heparin-induced thrombocytopenia. Blood 2000; 96: 1703-8.
  • 22 Cassel DL, Keller MA, Surrey S, Schwartz E, Schreiber AD, Rappaport EF, McKenzie SE. Differential expression of Fc gamma RIIA, Fc gamma RIIB and Fc gamma RIIC in hematopoietic cells: analysis of transcripts. Mol Immunol 1993; 30: 451-60.
  • 23 Warmerdam PA, van de Winkel JG, Gosselin EJ, Capel PJ. Molecular basis for a polymorphism of human Fc gamma receptor II (CD32). J Exp Med 1990; 172: 19-25.
  • 24 Warmerdam PA, van de Winkel JG, Vlug A, Westerdaal NA, Capel PJ. A single amino acid in the second Ig-like domain of the human Fc gamma receptor II is critical for human IgG2 binding. Immunol 1991; 147: 1338-43.
  • 25 Brandt JT, Isenhart CE, Osborne JM, Ahmed A, Anderson CL. On the role of platelet Fc gamma RIIa phenotype in heparin-induced thrombocytopenia. Thromb Haemost 1995; 74: 1564-72.
  • 26 Burgess JK, Lindeman R, Chesterman CN, Chong BH. Single amino acid mutation of Fc gamma receptor is associated with the development of heparin-induced thrombocytopenia. Br J Haematol 1995; 91: 761-6.
  • 27 Denomme GA, Warkentin TE, Horsewood P, Sheppard JA, Warner MN, Kelton JG. Activation of platelets by sera containing IgG1 heparin-dependent antibodies: an explanation for the predominance of the Fc gammaRIIa “low responder” (his131) gene in patients with heparin-induced thrombocytopenia. J Lab Clin Med 1997; 130: 278-84.
  • 28 Carlsson LE, Santoso S, Baurichter G, Kroll H, Papenberg S, Eichler P, Westerdaal NA, Kiefel V, van de Winkel JG, Greinacher A. Heparin-induced thrombocytopenia: new insights into the impact of the FcgammaRIIaR-H131 polymorphism. Blood 1998; 92: 1526-31.
  • 29 Pouplard C, Amiral J, Borg JY, Laporte-Simitsidis S, Delahousse B, Gruel Y. Decision analysis for use of platelet aggregation test, carbon 14-serotonin release assay, and heparin-platelet factor 4 enzyme-linked immunosorbent assay for diagnosis of heparin-induced thrombocytopenia. Am J Clin Pathol 1999; 111: 700-6.
  • 30 Sheridan D, Carter C, Kelton JG. A diagnostic test for heparin-induced thrombocytopenia. Blood 1986; 67: 27-30.
  • 31 Bell GI, Karam JH, Rutter WJ. Polymorphic DNA region adjacent to the 5′ end of the human insulin gene. Proc Natl Acad Sci USA 1981; 78: 5759-63.
  • 32 de Bruijne-Admiraal LG, Modderman PW, Von dem Borne AE, Sonnenberg A. P-selectin mediates Ca(2+)-dependent adhesion of activated platelets to many different types of leukocytes: detection by flow cytometry. Blood 1992; 80: 134-42.
  • 33 Bachelot C, Saffroy R, Gandrille S, Aiach M, Rendu F. Role of Fc gamma RIIA gene polymorphism in human platelet activation by monoclonal antibodies. Thromb Haemost 1995; 74: 1557-63.
  • 34 Amiral J, Marfaing-Koka A, Wolf M, Alessi MC, Tardy B, Boyer-Neumann C, Vissac AM, Fressinaud E, Poncz M, Meyer D. Presence of autoantibodies to interleukin-8 or neutrophil-activating peptide-2 in patients with heparin-associated thrombocytopenia. Blood 1996; 88: 410-6.
  • 35 Pidard D, Si-Tahar M, Chignard M. Neutrophil-Platelet interactions. In: Platelets, Thrombosis and the Vessel Wall. MC Berndt, ed. Advances In Vascular Biology Book Series. M Vadas, J Harlan, eds. Amsterdam: Harwood Academic Publishers; 2000: 189-207.
  • 36 de Gaetano G, Cerletti C, Evangelista V. Recent advances in platelet-polymorphonuclear leukocyte interaction. Haemostasis 1999; 29: 41-9.
  • 37 Larsen E, Celi A, Gilbert GE, Furie BC, Erban JK, Bonfanti R, Wagner DD, Furie B. PADGEM protein: a receptor that mediates the interaction of activated platelets with neutrophils and monocytes. Cell 1989; 59: 305-12.
  • 38 Hamburger SA, McEver RP. GMP-140 mediates adhesion of stimulated platelets to neutrophils. Blood 1990; 75: 550-4.
  • 39 Palabrica T, Lobb R, Furie BC, Aronovitz M, Benjamin C, Hsu YM, Sajer SA, Furie B. Leukocyte accumulation promoting fibrin deposition is mediated in vivo by P-selectin on adherent platelets. Nature 1992; 359: 848-51.
  • 40 Fareed J, Walenga JM, Hoppensteadt DA, Jeske WP, Lietz H, Ahmad S, Callas D, Messmore HL, Haas S. Selectins in the HIT syndrome: pathophysiologic role and therapeutic modulation. Semin Thromb Hemost 1999; 25 (Suppl. 01) 37-42.
  • 41 Jy W, Mao WW, Horstman LL, Valant PA, Ahn YS. A flow cytometric assay of platelet activation marker P-selectin (CD62P) distinguishes heparin-induced thrombocytopenia (HIT) from HIT with thrombosis (HITT). Thromb Haemost 1999; 82: 1255-9.
  • 42 Rinder CS, Bonan JL, Rinder HM, Mathew J, Hines R, Smith BR. Cardiopulmonary bypass induces leukocyte-platelet adhesion. Blood 1992; 79: 1201-5.
  • 43 Stewart GJ. Neutrophils and deep venous thrombosis. Haemostasis 1993; 23: 127-40.
  • 44 Siminiak T, Flores NA, Sheridan DJ. Neutrophil interactions with endothelium and platelets: possible role in the development of cardiovascular injury. Eur Heart J 1995; 16: 160-70.
  • 45 Ott I, Neumann FJ, Gawaz M, Schmitt M, Schomig A. Increased neutro-phil-platelet adhesion in patients with unstable angina. Circulation 1996; 94: 1239-46.
  • 46 Ikeda H, Ueyama T, Murohara T, Yasukawa H, Haramaki N, Eguchi H, Katoh A, Takajo Y, Onitsuka I, Ueno T, Tojo SJ, Imaizumi T. Adhesive interaction between P-selectin and sialyl Lewis(x) plays an important role in recurrent coronary arterial thrombosis in dogs. Arterioscler Thromb Vasc Biol 1999; 19: 1083-90.
  • 47 Gawaz MP, Mujais SK, Schmidt B, Gurland HJ. Platelet-leukocyte aggregation during hemodialysis. Kidney Int 1994; 46: 489-95.