Anticardiolipin and Anti-β2-GPI Are Two Distinct Populations of Autoantibodies

M Sorice; A Circella; T Griggi; T Garofalo; G Nicodemo; V Pittoni; G M Pontieri; L Lenti; G Valesini

doi:10.1055/s-0038-1650265

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1996; 75(02): 303-308
DOI: 10.1055/s-0038-1650265

Original Article

Schattauer GmbH Stuttgart

Anticardiolipin and Anti-β₂-GPI Are Two Distinct Populations of Autoantibodies

Authors

M Sorice

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
A Circella

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
T Griggi

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
T Garofalo

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
G Nicodemo

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
V Pittoni

¹Clinica Medica I, Universita “La Sapienza”, Rome, Italy
G M Pontieri

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
L Lenti

The Dipartimento di Medicina Sperimentale, Universita “La Sapienza”, Rome, Italy
G Valesini

¹Clinica Medica I, Universita “La Sapienza”, Rome, Italy

Abstract

PDF Download

Summary

This study has been undertaken to assess whether anticardiolipin and anti-(32-GPI are two distinct populations of (auto)antibodies, and to clarify whether the β₂-GPI region critical for phospholipid binding is also crucial for anti-β₂-GPI reactivity.

Fourteen of the 62 anticardiolipin (aCL) ELISA positive sera (22.6%) were positive for anti-β₂-GPI by immunoblotting, 42 (67.7%) for aCL using TLC immunostaining.

IgG fractions from 5 sera gave the same anticardiolipin reactivity detected by TLC immunostaining in the corresponding sera.

All anti-β₂-GPI-positive sera were reactive with the phenylthio-carbamyl derivative of the protein, indicating that binding of phe-nylisothiocyanate with lysine residues does not modify the molecule antigenicity. In addition, incubation of IgG fractions with the phospholipid binding site did not modify reactivity with β₂-GPI.

These findings demonstrate that: a) “true” antiphospholipid antibodies are detectable in patients sera; b) aCL and anti-β₂-GPI have a different immunological profile; c) the β₂-GPI phospholipid-binding site is not the region recognized by the antibodies.

PDF (446 kb)

References

References
1 Hughes GRV. The anticardiolipin syndrome. Clin Exp Rheumatol 1985; 3: 285-286
2 Hughes GRV, Harris EN, Gharavi AE. The anticardiolipin syndrome. J Rheumatol 1986; 13: 486-489
3 Harris EN, Gharavi AE, Loizou S. Crossreactivity of antiphospholipid antibodies. J Clin Lab Immunol 1985; 16: 1-6
4 Gharavi AE, Harris EN, Asherson RA, Hughes GV. Anticardiolipin antibodies: Isotype, distribution and phospholipid specificity. Ann Rheum Dis 1987; 46: 1-6
5 Galli M, Comfurius P, Maassen C, Hemker HC, De Baets MH, van Breda-Vriesman PJC, Barbui T, Zwaal RFA, Bevers EM. Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor. Lancet 1990; 355: 1544-1547
6 Me Neil HP, Simpson RJ, Chesterman CN, Krilis SA. Antiphospholipid antibodies are directed against a complex antigen that includes a lipid-binding inhibitor of coagulation: beta-2 glycoprotein-I (apolipoprotein II). Proc Natl Acad Sci USA 1990; 87: 4120-4124
7 Matsuura E, Igarashi Y, Fujimoto M, Ichikawa K, Koike T. Anticardiolipin cofactor(s) and differential diagnosis of autoimmune disease. Lancet 1990; 336: 177-178
8 Hunt JE, McNeil HP, Morgan GJ, Crameri RM, Kriis SA. A phospholipid-p2-glycoprotein I complex is an antigen for anticardiolipin antibodies occurring in autoimmune disease but not with infection. Lupus 1992; 1: 75-81
9 Hunt J, Krilis S. The fifth domain of b2-glycoprotein I contains a phospholipid binding site (Cys 281-Cys 288) and a region recognized by anticardiolipin antibodies. J Immunol 1994; 152: 653-659
10 Rauch J, Janoff AS. The nature of anti-phospholipid antibodies. J Rheumatol 1992; 19: 1782-1785
11 Viard JP, Amoura Z, Bach JF. Association of anti-(32 glycoprotein I antibodies with lupus-type circulating anticoagulant and thrombosis in systemic lupus erythematosus. Am J Med 1992; 93: 181-186
12 Roubey RAS, Pratt CW, Buyon JP, Winfield JB. Lupus anticoagulant activity of autoimmune antiphospholipid antibodies is dependent upon (32-glycoprotein I. J Clin Invest 1992; 90: 1100-1104
13 Galli M, Comfurius P, Barbui T, Zwaal RFA, Bevers EM. Anticoagulant activity of (32 glycoprotein I is potentiated by a distinct subgroup of anticardiolipin antibodies. Thromb Haemost 1992; 68: 297-300
14 Keeling DM, Wilson AJC, Mackie IJ, Isenberg DA, Machin SJ. Lupus anticoagulant activity of some antiphospholipid antibodies against phospholipid-bound (32 glycoprotein I. J Clin Pathol 1993; 46: 665-667
15 Sorice M, Griggi T, Circella A, Garofalo T, d’Agostino F, Pittoni V, Pon-tieri GM, Lenti L, Valesini G. Detection of antiphospholipid antibodies by immunostaining on thin layer chromatography plates. J Immunol Methods 1994; 173: 49-54
16 Tan EN, Cohen AS, Fries JF, Masi AT, McShane DJ, Rothfield NF, Shaller JG, Talal N, Winchester RG. The 1982 revised criteria for the classification of Systemic Lupus Erythematosus. Arthr Rheum 1982; 25: 1271-1277
17 Harris EN. The Second International Anti-Cardiolipin Standardization Workshop/The Kaps Group. Am J Clin Pathol 1990; 94: 476-484
18 Balestrieri G, Tincani A, Spatola L, Allegri F, Prati E, Cattaneo R, Valesini G, Del Papa N, Meroni P. Anti-beta2 glycoprotein I antibodies: a marker of antiphospholipid syndrome. Lupus 1995; 4: 122-130
19 Hardy RR. Purification and characterization of monoclonal antibodies. In: Handbook of Experimental Immunology 4th edn Weir DM. (ed) Oxford: Blackwell Scientific; 1986: 13-21
20 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970; 227: 680-685
21 Towbin H, Staehelin T, Gordon J. Elecrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
22 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phonol reagent. J Biol Chem 1951; 193: 265-275
23 Sammons DW, Adams LD, Nishizawa EE. Ultrasensitive silver-based color staining of polypeptides in polyacrylamide gels. Electrophoresis 1981; 2: 135-141
24 Stark GR. Recent developments in chemical modification and sequential degradation of protein. Adv Prot Chem 1970; 24: 261-308
25 Polz E, Kostner GM. The binding of beta2-gylcoprotein I to human serum lipoproteins distribution among density fraction. FEBS Lett 1979; 102: 183-186
26 Arvieux J, Roussel B, Jacob MC, Colomb MG. Measurement of anti-phospholipid antibodies by ELISA using beta 2-glycoprotein 1 as an antigen. J Immunol Methods 1991; 193: 223-229
27 Matsuura E, Igarashi Y, Fujimoto M, Ichikawa K, Suzuki T, Sumida T, Yasuda T, Koike T. Heterogeneity of anticardiolipin antibodies defined by the anticardiolipin cofactor. J Immunol 1992; 148: 3885-3891
28 Gharavi AE. Antiphospholipid cofactor. Stroke 1992; 23 (Suppl I) 1-10
29 Sammaritano LR, Lockshin MD, Gharavi AE. Antiphospholipid antibodies differ in aPL cofactor requirement. Lupus 1992; 1: 51-56
30 Valesini G, Shoenfeld Y. A new player in the antiphospholipid syndrome: the P2’GPI cofactor. Autoimmunity 1992; 14: 105-110
31 Keeling DM, Wilson AJC, Mackie IJ, Machin SJ, Isenberg DA. Some antiphospholipid antibodies’ bind to (32-glycoprotein I in the absence of phospholipid. Br J Haematol 1992; 82: 571-574
32 Pierangeli SS, Harris EN, Davis SA, DeLorenzo G. (32-glycoprotein I (p2GPI) enhanced cardiolipin binding activity but is not the antigen for antiphospholipid antibodies. Br J Haematol 1992; 82: 565-570
33 Arvieux J, Bensa JC, Roussel B, Colomb MG. Anti-|32-glycoprotein I antibodies. Clin Rev Allergy Immunol 1995; 13: 11-17
34 Pengo V, Biasiolo A, Fior MG. Autoimmune antiphospholipid antibodies are directed against a cryptic epitope expressed when p2-glycoprotein I is bound to a suitable surface. Thromb Haemost 1995; 73: 29-34
35 Roubey RAS, Eisenberg RA, Harper MF, Winfield JB. “Anticardiolipin” autoantibodies recognize (J2-glycoprotein I in the absence of phospholipid. Importance of Ag density and bivalent binding. J Immunol 1995; 154: 954-960
36 Arvieux BR, Ponard D, Colomb MG. IgG2 subclass restriction of anti-(32-glycoprotein I antibodies in autoimmune patients. Clin Exp Immunol 1994; 95: 310-315
37 Roubey RAS. Autoantibodies to phospholipid-binding plasma proteins: a new view of Lupus Anticoagulants and other “Antiphospholipid” autoantibodies. Blood 1994; 84: 2854-2867