Download PDF
Thromb Haemost 1995; 73(01): 029-034
DOI: 10.1055/s-0038-1653721
Original Article
Clinical Studies
Schattauer GmbH Stuttgart

Autoimmune Antiphospholipid Antibodies Are Directed against a Cryptic Epitope Expressed when β2-Glycoprotein I Is Bound to a Suitable Surface

Vittorio Pengo
The Dept. of Cardiology, Thrombosis Center, University of Padua School of Medicine, Padua, Italy
,
Alessandra Biasiolo
The Dept. of Cardiology, Thrombosis Center, University of Padua School of Medicine, Padua, Italy
,
Maria Grazia Fior
The Dept. of Cardiology, Thrombosis Center, University of Padua School of Medicine, Padua, Italy
› Author Affiliations
Further Information

Publication History

Received 08 June 1994

Accepted after resubmission 14 September 1994

Publication Date:
09 July 2018 (online)

  PDF Download  Permissions and Reprints

Summary

The antiphospholipid antibodies (aPL) present in autoimmune disorders are associated with thromboembolic episodes, and their binding to phospholipids (PL) is mediated by a plasma cofactor, β2-glycoprotein I (β2GPI). Both PL and β2GPI seem necessary for binding, thus indicating that the two components comprise the epitope against which aPL are directed. Using an anti-β2GPI antibody ELISA with the antigen adsorbed onto polyvinylchloride (PVC) plates, we detected high antibody titres in 12 out of 12 plasmas from patients with the antiphospholipid syndrome. No or very low positivity was obtained when the same ELISA was carried out in polystyrene (PST) plates, while an increasing positivity was found when processed (i.e. more hydrophilic) or COOH-surface PST plates were used. When (β2GPI dependent IgG-aPL were purified using agarose-immobilized cardio- lipin, 4 out of 4 preparations were highly positive in anti-β2GPI antibody ELISA using PVC plates, while β2GPI was not fully recognized by aPL-IgG when adsorbed onto PST plates. These findings demonstrate that aPL are, in fact, anti-β2GPI antibodies directed against a cryptic epitope which is expressed when β2GPI is bound to anionic phospholipid, or another suitable surface.

 

  • References

  • 1 Harris EN, Gharavi AE, Hughes GR V. Anti-phospholipid antibodies. Clin Rheum Dis 1985; 11: 591-609
    PubMedGoogle Scholar
  • 2 Gharavi AE, Harris EN, Asherson RA, Hughes GR V. Anticardiolipin anti bodies: isotype distribution and phospholipid specificity. Ann Rheum Dis 1987; 46: 1-6
    CrossrefPubMedGoogle Scholar
  • 3 Fiumara NJ. Biologic false positive reaction for syphilis. N Engl J Med 1963; 268: 402-405
    CrossrefPubMedGoogle Scholar
  • 4 Feinstein DI, Rapaport SI. Acquired inhibitors of blood coagulation. Prog Hemostas Thromb 1972; 1: 75-95
    PubMedGoogle Scholar
  • 5 Harris EN, Gharavi AE, Boey ML, Patel BM, Mackworth-Young CB, Loi-zou S, Hughes GR V. Anticardiolipin antibodies: detection by radioimmunoassay and association with thrombosis in systemic lupus erythematosus. Lancet 1983; 2: 1211-1214
    CrossrefPubMedGoogle Scholar
  • 6 Loizou S, McCrea JD, Rudge AC, Reynolds R, Boyle CC, Harris EN. Measurement of anticardiolipin antibodies by an enzyme-linked immunosorbent assay: standardization and quantitation of results. Clin Exp Immunol 1985; 62: 739-744
    PubMedGoogle Scholar
  • 7 Johansson AE, Lassus A. The occurrence of circulating anticoagulants in patients with syphilitic and biologically false positive antilipoidal antibodies. Ann Clin Res 1974; 6: 105-108
    PubMedGoogle Scholar
  • 8 Levy RA, Gharavi AE, Sammaritano LR, Habina L, Qamar T, Lockshin MD. Characteristics of IgG anitphospholipid antibodies in patients with systemic lupus erythematosus and syphilis. J Rheumatol 1990; 17: 1036-1041
    PubMedGoogle Scholar
  • 9 Costello PB, Green FA. Reactivtiy pattern of human anticardiolipin and other antiphospholipid antibodies in syphilitic sera. Infect Immunol 1986; 51: 771-775
    PubMedGoogle Scholar
  • 10 Santiago MB, Cossermelli W, Tuma MF, Pinto MN, Oliveira RM. Anticardiolipin antibodies in patients with infectious diseases. Clin Rheumatol 1989; 8: 23-28
    CrossrefPubMedGoogle Scholar
  • 11 McNeil HP, Chesterman CN, Krilis SA. Immunology and clinical importance of antiphospholipid antibodies. Adv Immunol 1991; 49: 193-280
    PubMedGoogle Scholar
  • 12 Brey RL, Coull BM. Antiphospholipid antibodies: origin, specificity, and mechansim of action. Stroke 1992; 23 suppl (Suppl. 01) I-15-I-18
    CrossrefPubMedGoogle Scholar
  • 13 Roubey RA S, Pratt CW, Buyon JP, Winfield JB. Lupus anticoagulant activity of autoimmune antiphopholipid antibodies is dependent upon β2-glycoprotein I. J Clin Invest 1992; 90: 1100-1104
    CrossrefPubMedGoogle Scholar
  • 14 Ordi J, Selva A, Monegal F, Procel JM, Martinez-Costa X, Vilardell M. Anticardiolipin antibodies and dependence of a serum cofactor. A mechanism of thrombosis J Rheumatol 1993; 20: 1321-1324
    PubMedGoogle Scholar
  • 15 McNeil HP, Simpson RJ, Chesterman CN, Krilis SA. Antiphospholipid antibodies are directed against a complex antigen that includes a lipid binding inhibitor of coagulation: β2-Glycoprotein-I (Apolipoprotein H). Proc Natl Acad Sci USA 1990; 87: 4120-4124
    CrossrefPubMedGoogle Scholar
  • 16 Galli M, Comfurius P, Maassen C, Hemker HC, De Baets MH, Van Brieda-Vriesman PJ C, Barbui T, Zwaal RF A, Bevers EM. Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor. Lancet 1990; 335: 1544-1547
    CrossrefPubMedGoogle Scholar
  • 17 Matsuura E, Igarashi Y, Fujimoto M, Ichikava K, Koike T. Anticardiolipin cofactor(s) and differential diagnosis of autoimmune disease. Lancet 1990; 336: 177-178
    PubMedGoogle Scholar
  • 18 Schultze HE, Heide K, Haupt H. Über ein bisher unbekanntes niedermolekulares β2-globulin des Humanserums. Naturwissenschaften 1961; 48: 719
    PubMedGoogle Scholar
  • 19 Viard JP, Amoura Z, Bach JF. Association of anti-β2-glycoprotein I antibodies with lupus-type circulating anticoagulant and thrombosis in systemic lupus erythematosus. Am J Med 1992; 93: 181-186
    CrossrefPubMedGoogle Scholar
  • 20 Wagenknecht DR, McIntyre JA. Changes in β2-glycoprotein I antigenicity induced by phospholipid binding. Thromb Haemost 1993; 69: 361-365
    Article in Thieme ConnectPubMedGoogle Scholar
  • 21 Rauch J, Janoff AS. The nature of antiphospholipid antibodies. J Rheumatol 1992; 19: 1782-1785
    PubMedGoogle Scholar
  • 22 Biasiolo A, Pengo V. Antiphospholipid antibodies are not present in the membrane of gel-filtered platelets of patients with IgG anticardiolipin antibodies, lupus anticoagulant and thrombosis. Blood Coag Fibrinol 1993; 4: 425-428
    CrossrefPubMedGoogle Scholar
  • 23 Harris EN, Gharavi AE, Patel SP, Hughes GV R. Evaluation of the anticardiolipin antibody test: report of an international workshop held 4 April 1986. Clin Exp Immunol 1987; 68: 215-222
    PubMedGoogle Scholar
  • 24 Thiagarajan P, Pengo V, Shapiro SS. The use of dilute Russell Viper Venom Time for the diagnosis of lupus anticoagulants. Blood 1986; 68: 869-873
    PubMedGoogle Scholar
  • 25 Triplett DA, Brandt JT, Kaczor D, Shaeffer 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
  • 26 Pengo V, Thiagarajan P, Shapiro SS, Heine MJ. Immunologie specificity and mechanism of action of IgG lupus anticoagulants. Blood 1987; 70: 69-76
    CrossrefPubMedGoogle Scholar
  • 27 Finlayson JS, Mushinki JF. Separation of subfractions of β2-glycoprotein I. Biochim Biophys Acta 1967; 147: 413-420
    CrossrefPubMedGoogle Scholar
  • 28 Pengo V, Ruffatti A, Baritussio A, De Marco L, Todesco S, Girolami A. Serum of normal subjects contains IgG with antibody-like specificity for phospholipids. Folia Haematol 1984; 111: 681-685
    PubMedGoogle Scholar
  • 29 Laemmli UK. Cleavage of structural proteins during the assembly of the head of Bacteriphage T4. Nature 1970; 227: 680-685
    CrossrefPubMedGoogle Scholar
  • 30 Pengo V, Biasiolo A. Purification of anticardiolipin and lupus anticoagulant activities by using cardiolipin immobilized on agarose beads. Thromb Res 1993; 72: 423-430
    CrossrefPubMedGoogle Scholar
  • 31 Pangborn MC. A new serologically active phospholipid from beef heart. Proc Soc Exp Biol (NY) 1941; 48: 484-486
    CrossrefPubMedGoogle Scholar
  • 32 Inoue K, Nojima S. Immunochemical studies of phospholipids. IV. The reactivities of antisera against natural cardiolipin and synthetic cardiolipin analogues-containing antigens. Chem Phys Lipids 1969; 3: 70-77
    CrossrefPubMedGoogle Scholar
  • 33 Moore JE, More CF. Biologically false positive serologic tests for syphilis: type, incidence and cause. J Chronic Dis 1952; 150: 467-473
    PubMedGoogle Scholar
  • 34 Hohmann A, Comacchio R, Boswarva V, Sutjita M, Bradley J. The H3 anti-phospholipid idiotype is found in patients with systemic lupus erythematosus (SLE) but not in patients with syphilis. Clin exp Immunol 1991; 86: 207-211
    PubMedGoogle Scholar
  • 35 Oosting JD, Derksen RH W M, Entjes HT I, Bouma BN, De Groot PG. Lupus anticoagulant activity is frequently dependent on the presence of β2-Glycoprotein I. Thromb Haemost 1992; 67: 499-502
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Galli M, Comfurius P, Barbui T, Zwaal RF A, Bevers EM. Anticoagulant activity of β2glycoprotein I is potentiated by a distinct subgroup of anticardiolipin antibodies. Thromb Haemost 1992; 68: 297-300
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Gharavi AE. Antiphospholipid cofactor. Stroke 1992; 23 suppl (Suppl. 01) I-7-I-10
    PubMedGoogle Scholar
  • 38 Verrier Jones J. Antiphospholipid antibodies: new perspective on antigenic specificity. J Rheumatol 1992; 19: 1774-1777
    PubMedGoogle Scholar
  • 39 Gharavi AZ, Sammaritano LR, Wen J, Elkon KB. Induction of antiphospholipid antibodies by immunization with β2-glycoprotein I (Apolipoprotein H). J Clin Invest 1992; 90: 1105-1109
    CrossrefPubMedGoogle Scholar
  • 40 Nimpf J, Bevers EM, Bomans PH H, Till U, Wurm H, Kostner GH, Zwaal RF A. Prothrombinase activity of human platelets is inhibited by β2-glyco-protein I. Biochim Biophys Acta 1986; 844: 142-149
    PubMedGoogle Scholar
  • 41 Rauch J, Meng Q-H, Tannebaum H. Lupus anticoagulant and antiplatelet properties of human hybridoma autoantibodies. J Immunol 1987; 139: 2598-2604
    PubMedGoogle Scholar
  • 42 Rauch J, Tannebaum H, Janoff AS. Distinguishing plasma lupus anticoagulants from anti-factor antibodies using hexagonal (II) phase phospholipids. Thromb Haemost 1989; 66: 453-458
    Article in Thieme ConnectPubMedGoogle Scholar
  • 43 Canfield WM, Kisiel W. Evidence of normal functional levels of activated protein C inhibitor in combined factor V/VIII deficiency disease. J Clin Invest 1982; 70: 1260-1272
    CrossrefPubMedGoogle Scholar
  • 44 Arvieux J, Roussel B, Jacob MC, Colomb MG. Measurement of anti-phospholipid antibodies by ELISA using β2-glycoprotein I as an antigen. J Immunol Meth 1991; 143: 223-229
    CrossrefPubMedGoogle Scholar
  • 45 Matsuura E, Igarashi Y, Yasuda T, Triplett DA, Koike T. Anticardiolipin antibodies recognize β2-glycoprotein I structure altered by interacting with an oxygen modified solid phase surface. J Exp Med 1994; 179: 457-462
    CrossrefPubMedGoogle Scholar
  • 46 Berger A, German JB, Gershwin ME. Implication of modifying cardiolipin acyl composition by diet. 1. Cardiolipin acyl chain is an important determinant in the binding to antiphospholipid antibodies in SLE sera. J Autoimmun 1992; 5: 229-241
    CrossrefPubMedGoogle Scholar
  • 47 Hasselaar P, Derksen RH W M, Blokzijl L, de Groot PG. Crossreactivity of antibodies directed against cardiolipin, DNA, endothelial cells and blood platelets. Thromb Haemost 1990; 63: 169-173
    Article in Thieme ConnectPubMedGoogle Scholar
  • 48 Bevers EM, Galli M, Barbui T, Comfurius P, Zwaal RF A. Lupus anticoagulant IgG’s (LA) are not directed to phospholipids only, but to a complex of lipid-bound human prothrombin. Thromb Haemost 1991; 66: 629-632
    Article in Thieme ConnectPubMedGoogle Scholar
  • 49 Fleck RA, Rapaport SI, Visajia Mohan Rao L. Anti-prothrombin antibodies and the lupus anticoagulant. Blood 1988; 72: 512-519
    CrossrefPubMedGoogle Scholar
  • 50 Janoff AS, Rauch J. The structural specificity of antiphospholipid antibodies in autoimmune disease. Chem Phys Lipids 1986; 40: 315-332
    CrossrefPubMedGoogle Scholar