Download PDF
Thromb Haemost 1993; 70(05): 787-793
DOI: 10.1055/s-0038-1649671
Coagulation
Schattauer GmbH Stuttgart

A Hexagonal (II) Phase Phospholipid Neutralization Assay for Lupus Anticoagulant Identification

Douglas A Triplett
The Department of Research Ball Memorial Hospital, Muncie, Indiana, USA
,
Linda K Barna
The Department of Research Ball Memorial Hospital, Muncie, Indiana, USA
,
Gail A Unger
The Department of Research Ball Memorial Hospital, Muncie, Indiana, USA
› Author Affiliations
Further Information

Publication History

Received 31 December 1992

Accepted after revision 09 June 1993

Publication Date:
05 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

Lupus anticoagulants (LAs) are immunoglobulins (IgG, IgM, or both) which interfere with in vitro phospholipid (PL) dependent tests of coagulation (e.g. APTT, dilute PT, dilute Russell Viper Venom Time). These antibodies may be identified in a wide variety of clinical settings. With the exception of heparinized patient samples, the presence of LAs is often the most common cause of an unexplained APTT in a routine clinical laboratory. The diagnosis of LAs is difficult due to variable screening reagent sensitivity and intrinsic heterogeneity of LAs. Recently, Rauch and colleagues have shown human monoclonal hybridoma LAs were inhibited by hexagonal (II) phase PLs. In contrast, lamellar phase PLs had no effect. We have evaluated a new assay system, Staclot LA®, which utilizes a hexagonal (II) phase PL (egg phosphatidylethanolamine [EPE]) as a confirmatory test for LAs. Plasma samples from the following patient populations were studied: LA positive, heparinized, oral anticoagulated, hemophilia A and B, and specific factor inhibitors (factors V, VIII, IX). Unlike previous studies, the LA positive patients were a mixed population including: autoimmune diseases, drug-induced, and post-infection. Our findings confirm the specificity of hexagonal (II) phase PL neutralization of LAs.

 

  • References

  • 1 Triplett DA, Brandt JT. Lupus anticoagulants: misnomer, paradox, riddle, epiphenomenon. Hematol Pathol 1988; 2: 121-123
    PubMedGoogle Scholar
  • 2 Love PE, Santoro SA. Antiphospholipid antibodies: anticardiolipin and the lupus anticoagulant in systemic lupus erythematosus (SLE) and in non-SLE disorders. Ann Int Med 1990; 112: 682-698
    CrossrefPubMedGoogle Scholar
  • 3 McNeil HP, Chesterman CN, Krilis SA. Immunology and clinical importance of antiphospholipid antibodies. Adv Immunol 1991; 49: 193-280
    PubMedGoogle Scholar
  • 4 Galli M, Comfurius P, Maassen C, Hemker HC, deBaets MH, van Breda-Vriesman PJC, Barbui T, Zwaal RFA, Bevers EM. Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma cofactor. Lancet 1990; 335: 1544-1547
    CrossrefPubMedGoogle Scholar
  • 5 McNeil HP, Simpson RJ, Chesterman CN, Krilis SA. Antiphospholipid antibodies are directed against a complex antigen that includes a lipid binding inhibitor of coagulation: B2 Glycoprotein I (apolipoprotein H). Proc Natl Acad Sci USA 1990; 87: 4120-4124
    CrossrefPubMedGoogle Scholar
  • 6 Matsuura E, Igarashi Y, Fujimoto M, Ichikawa K, Koike T. Anticardiolipin cofactor(s) and differential diagnosis of autoimmune disease. Lancet 1990; 336: 177-178
    PubMedGoogle Scholar
  • 7 Bevers EM, Galli M, Barbui T, Comfurius P, Zwaal RFA. Lupus anticoagulant IgG’s (LA) are not directed to phospholipids only but to a complex of lipid bound prothrombin. Thromb Haemost 1991; 66: 629-632
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Oosting JD, Derksen RHWM, Entjes HTI, Bouma BN, de Groot PG. Lupus anticoagulant activity is frequently dependent on the presence of B2 Glyoprotein I. Thromb Haemost 1992; 67: 499-502
    Article in Thieme ConnectPubMedGoogle Scholar
  • 9 Exner T, McRea J. Studies on the relationship between “antiphospholipid” antibodies and the lupus anticoagulant. Blood Coagulation and Fibrinolysis 1990; 1: 17-21
    CrossrefPubMedGoogle Scholar
  • 10 Triplett DA. Antiphospholipid antibodies and recurrent pregnancy loss. Am J Reprod Immunol 1989; 20: 52-67
    CrossrefPubMedGoogle Scholar
  • 11 Mannucci PM, Canciani MT, Mari D, Meucci P. The varied sensitivity of partial thromboplastin and prothrombin time reagents in the demonstration of lupus-like anticoagulant. Scand J Haematol 1979; 22: 423-432
    PubMedGoogle Scholar
  • 12 Triplett DA. Coagulation assays for the lupus anticoagulant: review and critique of current methodology. Stroke 1992; 23 (Suppl I): 111-114
    PubMedGoogle Scholar
  • 13 Exner T, Triplett DA, Taberner D, Machin SJ. Guidelines for testing and revised criteria for lupus anticoagulants. SSC Subcommittee for the Standardization of Lupus Anticoagulants. Thromb Haemost 1991; 65: 320-322
    Article in Thieme ConnectPubMedGoogle Scholar
  • 14 Schleider MA, Nachman RL, Jaffe EA, Coleman M. A clinical study of the lupus anticoagulant. Blood 1976; 48: 499-509
    PubMedGoogle Scholar
  • 15 Thiagarajan P, Pengo V, Shapiro SS. The use of the dilute Russell viper venom time for the diagnosis of lupus anticoagulants. Blood 1986; 68: 869-874
    PubMedGoogle Scholar
  • 16 Alving BM, Barr CF, Johansen LE, Tang DB. Comparison between a one-point dilute phospholipid APTT and the dilute Russel viper venom time for verification of lupus anticoagulants. Thromb Haemost 1992; 67: 672-678
    Article in Thieme ConnectPubMedGoogle Scholar
  • 17 Triplett DA, Brandt JT, Kaczor D, Schaeffer J. Laboratory diagnosis of lupus inhibitors: a comparison of the tissue thromboplastin inhibition procedure with a new platelet neutralization procedure. Am J Clin Pathol 1983; 79: 678-682
    CrossrefPubMedGoogle Scholar
  • 18 Rosove MH, Ismail M, Kozoil BJ, Runge A, Kasper CK. Lupus anticoagulants: improved diagnosis with a kaolin clotting time using rabbit brain phospholipid in standard and high concentrations. Blood 1986; 68: 472-478
    PubMedGoogle Scholar
  • 19 Arnout J, Huybrechts E, Vanrusselt E, Vermylen J. A new lupus anticoagulant neutralization test based on platelet-derived vesicles. Br J Haematol 1992; 80: 341-346
    CrossrefPubMedGoogle Scholar
  • 20 Rauch J, Tannenbaum M, Tannenbaum H, Ramelson H, Cullis PR, Tilcock CPS, Hope MJ, Janoff AS. Human hybridoma lupus anticoagulants distinguish between lamellar and hexagonal phase lipid systems. J Biol Chem 1986; 261: 9672-9677
    PubMedGoogle Scholar
  • 21 Janoff AS, Rauch J. The structural specificity of antiphospholipid antibodies in autoimmune disease. Chem Phys Lipids 1986; 40: 315-332
    CrossrefPubMedGoogle Scholar
  • 22 Rauch J, Tannenbaum M, Janoff AS. Distinguishing plasma lupus anticoagulants from anti-factor antibodies using hexagonal (II) phase phospholipids. Thromb Haemost 1989; 62: 892-896
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Rauch J, Janoff AS. Phospholipid in the hexagonal II phase is immunogenic. Evidence for immuno recognition of nonbilayer lipid phases in vivo. Proc Natl Acad Sci USA 1990; 87: 4112-4114
    CrossrefPubMedGoogle Scholar
  • 24 Triplett DA, Harms CS. Procedures for the coagulation laboratory. American Society of Clinical Pathologists. Chicago: Illinois; 1981
    Google Scholar
  • 25 Triplett DA, Hannah D, Barna L. Hexagonal (II) phase phospholipid neutralization of lupus anticoagulants: a specific and sensitive confirmatory procedure. Thromb Haemost 1991; 65: 1255 (abstract 2021)
    PubMedGoogle Scholar
  • 26 Triplett DA, Brandt JT, Musgrave KA, Orr CA. The relationship between lupus anticoagulants and antibodies to phospholipid. JAMA 1988; 259: 550-554
    CrossrefPubMedGoogle Scholar
  • 27 Triplett DA. Screening for the lupus anticoagulant. Res Clin Lab 1989; 19: 379-389
    PubMedGoogle Scholar
  • 28 National Committee for Clinical Laboratory Standards. Collection, Transport, and Processing of Blood Specimens for Coagulation Testing and Performance of Coagulation Assays. 2nd. Edition Approved Guidelines NCCLS Document H21-A2, NCCLS. Villanova; Pennsylvania: 1991
    Google Scholar
  • 29 Roisin JP, Contant G, Martinoli JL. Detection of lupus-like anticoagulants (LA). Use of hexagonal phosphatidylethanolamine and of an APTT reagent sensitive to LA. Thromb Haemost 1991; 65: 1255 (abstract 2023)
    PubMedGoogle Scholar
  • 30 Triplett DA, Brandt JT, Maas RL. The laboratory heterogeneity of lupus anticoagulants. Arch Pathol Lab Med 1985; 109: 946-951
    PubMedGoogle Scholar
  • 31 Vermylen J, Arnout J. Is the antiphospholipid syndrome caused by antibodies directed against physiologically relevant phospholipid-protein complexes?. J Lab Clin Med 1992; 120: 10-12 (editorial)
    PubMedGoogle Scholar
  • 32 Galli M, Comfurius P, Barbui T, Zwaal RFA, Bevers EM. Anticoagulant activity of B2 Glycoprotein I is potentiated by a distinct subgroup of anticardiolipin antibodies. Thromb Haemost 1992; 68: 297-300
    Article in Thieme ConnectPubMedGoogle Scholar
  • 33 Bevers EM, Galli M. Cofactors involved in the antiphospholipid syndrome. Lupus 1992; 1: 51-53 (editorial)
    CrossrefPubMedGoogle Scholar
  • 34 Margolius Jr A, Jackson DP, Ratnoff OD. Circulating anticoagulants; a study of 40 cases and review of the literature. Med 1961; 40: 145-202
    CrossrefPubMedGoogle Scholar
  • 35 Zehnder JL, Leung LLK. Development of antibodies to thrombin and factor V with recurrent bleeding in a patient exposed to topical bovine thrombin. Blood 1990; 76: 2011-2016
    PubMedGoogle Scholar
  • 36 Nichols WL, Daniels TM, Fisher PK, Owen WG, Figueroa PI, Pineda AA. Inhibitors of coagulation factor V and thrombin associated with surgical use of topical bovine thrombin or fibrin “glue”. Blood 1991; 78 (Suppl. 01) 63a (abstract 241)
    PubMedGoogle Scholar
  • 37 Spero JA, Triplett DA, Cmolik BL, Reid C, Clark RE. A family of clotting factor inhibitors primarily involving FV and FX1 occurring after cardiovascular and neurosurgical procedures. Blood 1991; 78 (Suppl. 01) 63a (abstract 243)
    PubMedGoogle Scholar
  • 38 Rapaport SI, Zivelin A, Minow RA, Hunter CS, Donnelly K. Clinical significance of antibodies to bovine and human thrombin and factor V after surgical use of bovine thrombin. Am J Clin Pathol 1992; 97: 84-91
    CrossrefPubMedGoogle Scholar
  • 39 Brandt JT, Britton A, Kraut EA. Spontaneous factor V inhibitor with unexpected laboratory features. Arch Pathol Lab Med 1986; 110: 224-227
    PubMedGoogle Scholar
  • 40 McGlasson DL, Mercier RJ, Potema LF, Patterson WR. Platelet neutralization procedure: atypical results seen with factor V deficiency with and without an inhibitor. Clin Lab Sci 1990; 3: 119-121
    PubMedGoogle Scholar
  • 41 LeFrère JJ, Gozin D, Modai J, Vitteloq D. Circulating anticoagulant in the acquire immunodeficiency syndrome. Ann Int Med 1987; 107: 429-430 (letter)
    PubMedGoogle Scholar
  • 42 Haire WD. The acquired immunodeficiency syndrome and lupus anticoagulant. JAMA 1984; 252: 3399
    CrossrefPubMedGoogle Scholar
  • 43 Barrowcliffe TW, Kemball-Cook G, Gray E. Binding to phospholipid protects factor VIII from inactivation by human antibodies. J Lab Clin Med 1983; 101: 34-43
    PubMedGoogle Scholar
  • 44 Kemball-Cook G, Barrowcliffe TW. Interaction of factor VIII with phospholipids: role of composition and negative charge. Thromb Res 1992; 67: 57-71
    CrossrefPubMedGoogle Scholar