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DOI: 10.1055/s-0037-1615190
Heparin-induced Thrombocytopenia: IgG Binding to PF4-Heparin Complexes in the Fluid Phase and Cross-reactivity with Low Molecular Weight Heparin and Heparinoid
Publication History
Received
28 November 1997
Accepted after revision
02 April 1998
Publication Date:
08 December 2017 (online)
Summary
Early diagnosis of heparin-induced thrombocytopenia (HIT) is essential to reduce morbidity and mortality. We report an enzyme immunoassay which detects the binding of HIT IgG to PF4-heparin in the fluid phase. Our fluid phase assay produces consistently low background and can detect low levels of anti-PF4-heparin. It is suited to testing alternative anticoagulants because, unlike in an ELISA, a clearly defined amount of antigen is available for antibody binding. We were able to detect anti-PF4-heparin IgG in 26/28 (93%) HIT patients. We investigated cross-reactivity of anti-PF4-heparin antibodies with PF4 complexed to alternative heparin-like anticoagulants. Low molecular weight heparins cross-reacted with 23/26 (88%) of the sera from HIT patients while half of the HIT sera weakly cross-reacted with PF4-danaparoid (Orgaran). The thrombocytopenia and thrombosis of most of these patients resolved during danaparoid therapy, indicating that detection of low affinity antibodies to PF4-danaparoid by immunoassay may not be an absolute contraindication for danaparoid administration.
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References
- 1 Chong BH. Heparin-induced thrombocytopenia. Br J Haematol 1995; 89: 431-9.
- 2 Sheridan D, Carter C, Kelton JG. A diagnostic test for heparin-induced thrombocytopenia. Blood 1986; 67: 27-30.
- 3 Chong BH, Burgess J, Ismail F. The clinical usefulness of the platelet aggregation test for the diagnosis of heparin-induced thrombocytopenia. Thromb Haemost 1993; 69: 344-50.
- 4 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.
- 5 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.
- 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 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.
- 8 Faivre R, Kieffer Y, Bassand JP, Maurat JP. Severe thrombopenia from heparin: Value of the use of low molecular weight heparin. Apropos of 6 cases. Arch Mal Coeur Vaiss 1985; 78: 27-30 (Abstract).
- 9 Gouault-Heilmann M, Huet Y, Adnot S, Contant G, Bonnet F, Intrator L, Payen D, Levent M. Low molecular weight heparin fractions as an alternative therapy in heparin-induced thrombocytopenia. Haemostasis 1987; 17: 134-40.
- 10 Farag SS, Savoia H, Omalley CJ, McGrath KM. Lack of in vitro cross-reactivity predicts safety of low-molecular weight heparins in heparin-induced thrombocytopenia. Clinical & Applied Thrombosis-Hemostasis 1997; 3: 58-62.
- 11 Makhoul RG, Greenberg CS, McCann RL. Heparin-associated thrombocytopenia and thrombosis: a serious clinical problem and potential solution. J Vasc Surg 1986; 4: 522-8.
- 12 Ramakrishna R, Manoharan A, Kwan YL, Kyle PW. Heparin-induced thrombocytopenia: cross-reactivity between standard heparin, low molecular weight heparin, dalteparin (Fragmin) and heparinoid, danaparoid (Orgaran). Br J Haematol 1995; 91: 736-8.
- 13 Greinacher A, Drost W, Michels I, Leitl J, Gottsmann M, Kohl HJ, Glaser M, Mueller-Eckhardt C. Heparin-associated thrombocytopenia: successful therapy with the heparinoid Org 10172 in a patient showing cross-reaction to LMW heparins. Ann Hematol 1992; 64: 40-2.
- 14 Horellou MH, Conard J, Lecrubier C, Samama M, Roque-D’Orbcastel O, de Fenoyl O, Di Maria G, Bernadou A. Persistent heparin induced thrombocytopenia despite therapy with low molecular weight heparin. Thromb Haemost 1984; 51: 134.
- 15 Leroy J, Leclerc MH, Delahousse B, Guerois C, Foloppe P, Gruel Y, Toulemonde F. Treatment of heparin-associated thrombocytopenia and thrombosis with low molecular weight heparin (CY 216). Semin Thromb Hemost 1985; 11: 326-9.
- 16 Bauer TL, Arepally G, Konkle BA, Mestichelli B, Shapiro SS, Cines DB, Poncz M, Mcnulty S, Amiral J, Hauck WW, Edie RN, Mannion JD. Prevalence of heparin-associated antibodies without thrombosis in patients undergoing cardiopulmonary bypass surgery. Circulation 1997; 95: 1242-6.
- 17 Arepally G, Reynolds C, Tomaski A, Amiral J, Jawad A, Poncz M, Cines DB. Comparison of PF4/heparin ELISA assay with the 14C-serotonin release assay in the diagnosis of heparin-induced thrombocytopenia. Am J Clin Pathol 1995; 104: 648-54.
- 18 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.
- 19 Horne 3rd MK, Alkins BR. Platelet binding of IgG from patients with heparin-induced thrombocytopenia. J Lab Clin Med 1996; 127: 435-42.
- 20 Wilchek M, Ben-Hur H, Bayer EA. p-Diazobenzoyl biocytin – A new biotinylating reagent for the labeling of tyrosines and histidines in proteins. Biochem Biophys Res Commun 1986; 138: 872-9.
- 21 Ngai PK, Ackermann F, Wendt H, Savoca R, Bosshard HR. Protein A antibody-capture ELISA (PACE): an ELISA format to avoid denaturation of surface-adsorbed antigens. J Immunol Methods 1993; 158: 267-76.
- 22 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.
- 23 Warkentin TE, Elavathil LJ, Hayward CP, Johnston MA, Russett JI, Kelton JG. The pathogenesis of venous limb gangrene associated with heparin-induced thrombocytopenia. Ann Intern Med 1997; 127: 804-12.
- 24 Potzsch B, Unkrig C, Madlener K, Greinacher A, Muller-Berghaus G. APC resistance and early onset of oral anticoagulation are high thrombotic risk factors in patients with heparin-associated thrombocytopenia (HAT). Ann Hematol 1996; 72 (Suppl. 01) A6 (Abstract).
- 25 Amiral J, Marfaingkoka A, Wolf M, Alessi MC, Tardy B, Boyerneumann C, Vissac AM, Fressinaud E, Poncz M, Meyer D. Presence of autoanti-bodies to interleukin-8 or neutrophil-activating peptide-2 in patients with heparin-associated thrombocytopenia. Blood 1996; 88: 410-6.
- 26 Vun CM, Evans S, Chong BH. Cross-reactivity study of low molecular weight heparins and heparinoid in heparin-induced thrombocytopenia. Thromb Res 1996; 81: 525-32.
- 27 Slocum MM, Adams Jr. JG, Teel R, Spadone DP, Silver D. Use of enoxaparin in patients with heparin-induced thrombocytopenia syndrome. J Vasc Surg 1996; 23: 839-43.
- 28 Greinacher A, Michels I, Kiefel V, Mueller-Eckhardt C. A rapid and sensitive test for diagnosing heparin-associated thrombocytopenia. Thromb Haemost 1991; 66: 734-6.
- 29 Greinacher A, Michels I, Mueller-Eckhardt C. Heparin-associated thrombocytopenia: the antibody is not heparin specific. Thromb Haemost 1992; 67: 545-9.
- 30 Greinacher A, Michels I, Liebenhoff U, Presek P, Mueller-Eckhardt C. Heparin-associated thrombocytopenia: immune complexes are attached to the platelet membrane by the negative charge of highly sulphated oligosaccharides. Br J Haematol 1993; 84: 711-6.
- 31 Kikta MJ, Keller MP, Humphrey PW, Silver D. Can low molecular weight heparins and heparinoids be safely given to patients with heparin-induced thrombocytopenia syndrome?. Surgery 1993; 114: 705-10.
- 32 Luzzatto G, Cordiano I, Patrassi G, Fabris F. Heparin-induced thrombocytopenia: discrepancy between the presence of IgG cross-reacting in vitro with fraxiparine and its successful clinical use. Thromb Haemost 1995; 74: 1607-9.
- 33 Tardy Poncet B, Mahul P, Beraud AM, Favre JP, Tardy B, Guyotat D. Failure of Orgaran therapy in a patient with a previous heparin-induced thrombocytopenia syndrome. Br J Haematol 1995; 90: 969-70.