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Thromb Haemost 2001; 86(06): 1475-1482
DOI: 10.1055/s-0037-1616751
Review Article
Schattauer GmbH

Effect of Cardiolipin Oxidation on Solid-Phase Immunoassay for Antiphospholipid Antibodies

Michael Schlame
1   Department of Anesthesiology and the Division of Rheumatic Diseases, Hospital for Special Surgery, and the Biopolymer Mass Spectrometry Core Facility, Weill Medical College of Cornell University, New York, USA
,
Ivan Haller
1   Department of Anesthesiology and the Division of Rheumatic Diseases, Hospital for Special Surgery, and the Biopolymer Mass Spectrometry Core Facility, Weill Medical College of Cornell University, New York, USA
,
Lisa R. Sammaritano
1   Department of Anesthesiology and the Division of Rheumatic Diseases, Hospital for Special Surgery, and the Biopolymer Mass Spectrometry Core Facility, Weill Medical College of Cornell University, New York, USA
,
Thomas J. J. Blanck
1   Department of Anesthesiology and the Division of Rheumatic Diseases, Hospital for Special Surgery, and the Biopolymer Mass Spectrometry Core Facility, Weill Medical College of Cornell University, New York, USA
› Author Affiliations
Further Information

Publication History

Received 30 April 2000

Accepted 15 April 2000

Publication Date:
12 December 2017 (online)

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Summary

Diagnostic assays for antiphospholipid antibodies are routinely performed on microtitre plates coated with cardiolipin. Here we show that contact between cardiolipin and NUNC-Immuno® plates leads to extensive oxidation, generating a series of peroxy-cardiolipins which were identified by electrospray ionization mass spectrometry. To investigate the impact of oxidation on the antibody assay, cardiolipin was resolved into 12 molecular species, including oxidized species and non-oxidized species with different degrees of unsaturation. All 12 species reacted under anaerobic conditions with serum from patients with primary antiphospholipid syndrome. Immune reactivity was similar for tetralinoleoyl-cardiolipin, trilinoleoyl-oleoyl-cardiolipin, and peroxycardiolipins, but somewhat lower for tristearoyl-oleoyl-cardiolipin. Oxidative treatment of cardiolipin with air, cytochrome c, or Cu2+/tert-butylhydroperoxide, either before or during the assay, did not enhance immune reactivity. Similar results were obtained with a monoclonal IgM from lupus-prone mice, that binds cardiolipin in the absence of protein cofactors. We conclude that the solid-phase assay for antiphospholipid antibodies can be supported by various oxidized and nonoxididized molecular species of cardiolipin.

Keywords

Anticardiolipin antibodies - antigen specificity - antiphospholipid syndrome - enzyme-linked immunoassay - lipid oxidation
 

  • References

  • 1 Alarcon-Segovia D, Cabral AR. The concept and classification of antiphospholipid/cofactor syndromes. Lupus 1996; 5: 364-7.
    CrossrefPubMedGoogle Scholar
  • 2 Branch DW, Silver RM. Criteria for antiphospholipid syndrome: Early pregnancy loss, fetal loss or recurrent pregnancy loss?. Lupus 1996; 5: 409-13.
    CrossrefPubMedGoogle Scholar
  • 3 Wilson WA, Gharavi AE, Koike T, Lockshin MD, Branch DW, Piette JC, Brey R, Derksen R, Harris EN, Hughes GRV, Triplett DA, Khamashta MA. International concensus statement on preliminary classification criteria for definite antiphospholipid syndrome: report of an international workshop. Arthritis Rheum 1999; 42: 1309-11.
    CrossrefPubMedGoogle Scholar
  • 4 Reyes H, Dearing L, Shoenfeld Y, Peter JB. Antiphospholipid antibodies: A critique of their heterogeneity and hegemony. Sem Thromb Hemost 1994; 20: 89-100.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Kandiah DA, Sali A, Sheng Y, Victoria EJ, Marquis DM, Coutts SM, Krilis SA. Current insights into the “antiphospholipid” syndrome: clinical, immunological, and molecular aspects. Adv Immunol 1998; 70: 507-63.
    PubMedGoogle Scholar
  • 6 Iverson GM, Victoria EJ, Marquis DM. Anti-β2 glycoprotein I (β2GPI) autoantibodies recognize an epitope on the first domain of β2GPI. Proc Natl Acad Sci USA 1998; 95: 15542-46.
    CrossrefPubMedGoogle Scholar
  • 7 Tincani A, Spatola L, Prati E, Allegri F, Ferremi P, Cattaneo R, Meroni P, Balestrieri G. The anti-beta2-glycoprotein I activity in human anti-phospholipid syndrome sera is due to monoreactive low-affinity autoantibodies directed to epitopes located on native beta2-glycoprotein I and preserved during species’ evolution. J Immunol 1996; 157: 5732-8.
    PubMedGoogle Scholar
  • 8 Roubey RA, Eisenberg RA, Harper MF, Winfield JB. “Anticardiolipin” autoantibodies recognize beta 2-glycoprotein I in the absence of phospholipid. Importance of Ag density and bivalent binding. J Immunol 1995; 154: 954-60.
    PubMedGoogle Scholar
  • 9 Hörkkö S, Miller E, Dudl E, Reaven P, Curtiss LK, Zvaifler NJ, Terkeltaub R, Pierangeli SS, Branch DW, Palinski W, Witztum JL. Antiphospholipid antibodies are directed against epitopes of oxidized phospholipids. Recognition of cardiolipin by monoclonal antibodies to epitopes of oxidized low density lipoprotein. J Clin Invest 1996; 98: 815-25.
    CrossrefPubMedGoogle Scholar
  • 10 Hörkkö S, Miller E, Branch DW, Palinski W, Witztum JL. The epitopes for some antiphospholipid antibodies are adducts of oxidized phospholipid and β2 glycoprotein 1 (and other proteins). Proc Natl Acad Sci USA 1997; 94: 10356-61.
    CrossrefPubMedGoogle Scholar
  • 11 Fickl H, Van Antwerpen VL, Richards GA, Van der Westhuyzen DR, Davies N, Van der Walt R, Van der Merwe CA, Anderson R. Increased levels of autoantibodies to cardiolipin and oxidised low density lipoprotein are inversely associated with plasma vitamin C status in cigarette smokers. Atherosclerosis 1996; 124: 75-81.
    CrossrefPubMedGoogle Scholar
  • 12 Iuliano L, Pratico D, Ferro D, Pittoni V, Valesini G, Lawson J, FitzGerald GA, Violi F. Enhanced lipid peroxidation in patients positive for antiphospholipid antibodies. Blood 1997; 90: 3931-5.
    CrossrefPubMedGoogle Scholar
  • 13 Vaarala O, Palosuo T, Kleemola M, Aho K. Anticardiolipin response in acute infections. Clin Immunol Immunopathol 1986; 41: 8-15.
    CrossrefPubMedGoogle Scholar
  • 14 Pereira B, Benedict CR, Le A, Shapiro SS, Thiagarajan P. Cardiolipin binding a light chain from lupus-prone mice. Biochemistry 1998; 37: 1430-7.
    CrossrefPubMedGoogle Scholar
  • 15 Iwase H, Takatori T, Nagao M, Iwadate K, Nakajima M. Monoepoxide production from linoleic acid by cytochrome c in the presence of cardiolipin. Biochem Biophys Res Comm 1996; 222: 83-9.
    CrossrefPubMedGoogle Scholar
  • 16 Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 1959; 37: 911-7.
    CrossrefPubMedGoogle Scholar
  • 17 Teng JI, Smith LL. High-performance liquid chromatography of cardiolipin. J Chromatogr 1985; 339: 35-44.
    CrossrefPubMedGoogle Scholar
  • 18 Schlame M, Rua D, Greenberg M. The biosynthesis and functional role of cardiolipin. Progr Lipid Res 2000; 39: 257-88.
    CrossrefPubMedGoogle Scholar
  • 19 Tincani A, Balestrieri G, Spatola L, Cinquini M, Meroni PL, Roubey RA. Anticardiolipin and anti-β2 glycoprotein I immunoassays in the diagnosis of antiphospholipid syndrome. Clin Exp Rheumatol 1998; 16: 396-402.
    PubMedGoogle Scholar
  • 20 Firer MA, Spivak T, Shoenfeld Y, Slor H. The effects of incubation temperature and coating procedure on the measurement of antibodies to cardiolipin. J Immunol Meth 1991; 143: 31-9.
    CrossrefPubMedGoogle Scholar
  • 21 Hughes JR, Davies A, Prentice CRM. The effect of different microtitre plates on the enzyme-linked immunosorbent assay (ELISA) for anticardiolipin antibodies (ACAs). Thromb Res 1996; 84: 217-22.
    CrossrefPubMedGoogle Scholar
  • 22 Kilpatrick DC. Factors affecting cardiolipin antibody assays: modification with polyethylene glycol compound. Br J Haematol 1998; 100: 52-7.
    CrossrefPubMedGoogle Scholar
  • 23 Matsuda J, Gohchi K, Gotoh M, Kawasugi K, Katoh M. β2-Glycoprotein I-dependent anticardiolipin antibody is reactive to both β2-glycoprotein I/oxidized as well as β2-glycoprotein I/reduced cardiolipin. Thromb Res 1999; 95: 63-7.
    CrossrefPubMedGoogle Scholar
  • 24 Levy RA, Gharavi AE, Sammaritano LR, Habina L, Lockshin MD. Fatty acid chain is a critical epitope for antiphospholipid antibody. J Clin Immunol 1990; 10: 141-5.
    CrossrefPubMedGoogle Scholar
  • 25 Palinski W, Tangirala RK, Miller E, Young SG, Witztum JL. Increased autoantibody titers against epitopes of oxidized low density lipoprotein in LDL receptor-deficient mice with increased atherosclerosis. Arterioscler Thromb Vasc Biol 1995; 15: 1569-76.
    CrossrefPubMedGoogle Scholar
  • 26 Hörkkö S, Bird DA, Miller E, Itabe H, Leitinger N, Subbanagounder G, Berliner JA, Friedman P, Dennis EA, Curtiss LK, Palinski W, Witztum JL. Monoclonal autoantibodies specific for oxidized phospholipids or oxidized phospholipid-protein adducts inhibit macrophage uptake of oxidized low-density lipoproteins. J Clin Invest 1999; 103: 117-28.
    CrossrefPubMedGoogle Scholar
  • 27 Berger A, German JB, Gershwin ME. Implications of modifying cardiolipin acyl composition by diet. I. Cardiolipin acyl chain is an important determinant in the binding to antiphospholipid in SLE sera. J Autoimmun 1992; 5: 229-41.
    CrossrefPubMedGoogle Scholar
  • 28 Gutteridge JMC, Halliwell B. The measurement and mechanism of lipid peroxidation in biological systems. Trends Biol Sci 1990; 15: 129-35.
    CrossrefPubMedGoogle Scholar
  • 29 Rauch J, Janoff AS. The nature of antiphospholipid antibodies. J Rheumatol 1992; 19: 1782-85.
    PubMedGoogle Scholar
  • 30 Sankaram MB, Powell GL, Marsh D. Effect of acyl chain composition on salt-induced lamellar to inverted hexagonal phase transitions in cardiolipin. Biochim Biophys Acta 1989; 980: 389-92.
    CrossrefPubMedGoogle Scholar
  • 31 Bartlett GR. Phosphorus assay in column chromatography. J Biol Chem 1959; 234: 466-8.
    PubMedGoogle Scholar