The presence of antibodies against the complex of prothrombin and phosphatidylserine (aPS/PT) more significantly correlates with manifestations of antiphospholipid syndrome (APS) and with the presence of lupus anticoagulants (LA) than antibodies against prothrombin bound to oxygenated polystyrene (aPT-A). To investigate immunological specificities and functional activities of aPS/PT, four monoclonal aPS/PT, designated as HG-4, KE-6, KF-5 and KF-6, from two patients with antiphospholipid antibodies (aPL) were established and characterized. Three of these antibodies (HG-4, KF-5 and KF-6) recognized the complex of phosphatidylserine and prothrombin, but did not react to prothrombin directly coated on oxygenated plates. KE-6 bound not only to the complex of phosphatidylserine and prothrombin but also to prothrombin on oxygenated plates. None of them showed the binding activity to prothrombin directly coated on non-oxygenated plates. HG-4, KE-6 and KF-5 had LA-like activity. The findings support the hypothesis that autoimmune aPS/PT recognize the cryptic epitopes or neoepitopes exposed upon interaction between prothrombin and phosphatidylserine, and that aPS/PT are, at least in part, responsible for LA activity. Lupus (2007) 16, 509—516.
Hughes GRThe antiphospholipid syndrome: ten years on. Lancet1993; 342(8867): 341—344.
2.
Khamashta MA , Hughes GRAntiphospholipid antibodies and antiphospholipid syndrome. Curr Opin Rheumatol1995; 7(5): 389—394.
3.
Galli M., Comfurius P., Maassen C. et al. Anticardiolipin antibodies (ACA) directed not to cardiolipin but to a plasma protein cofactor. Lancet1990 ; 335(8705): 1544—1547.
4.
Matsuura E., Igarashi Y., Fujimoto M., Ichikawa K., Koike T.Anticardiolipin cofactor(s) and differential diagnosis of autoimmune disease . Lancet1990; 336(8708): 177—178.
5.
McNeil HP, Simpson RJ, Chesterman CN, Krilis SAAnti-phospholipid antibodies are directed against a complex antigen that includes a lipid-binding inhibitor of coagulation: beta 2-glycoprotein I (apolipoprotein H). Proc Natl Acad Sci USA1990; 87(11): 4120—4124.
6.
Roubey RAImmunology of the antiphospholipid antibody syndrome. Arthritis Rheum1996; 39(9): 1444—1454.
7.
Matsuura E., Igarashi Y., Yasuda T., Triplett DA, Koike T.Anticardiolipin antibodies recognize beta 2-glycoprotein I structure altered by interacting with an oxygen modified solid phase surface. J Exp Med1994; 179(2): 457—462.
8.
Oosting JD, Derksen R., Entjes H., Bouma B., de Groot P.Lupus anticoagulant activity is frequently dependent on the presence of beta-2 glycoprotein I. Thromb Haemost1992 ; 67: 499—502.
9.
Ichikawa K., Khamashta MA, Koike T., Matsuura E., Hughes GRbeta 2-Glycoprotein I reactivity of monoclonal anticardiolipin antibodies from patients with the antiphospholipid syndrome. Arthritis Rheum1994; 37(10): 1453—1461.
10.
Takeya H., Mori T., Gabazza EC et al. Anti-beta2-glycoprotein I (beta2GPI) monoclonal antibodies with lupus anticoagulant-like activity enhance the beta2GPI binding to phospholipids . J Clin Invest1997; 99(9): 2260—2268.
11.
Löliger A.Prothrombin as a co-factor of the circulating anticoagulant in systemic lupus erythematosus?Thromb Diath Haemorrh1959; 3: 237—256.
12.
Galli M., Barbui T.Antiprothrombin antibodies: detection and clinical significance in the antiphospholipid syndrome. Blood1999; 93(7): 2149—2157.
13.
Wu JR, Lentz BRPhospholipid-specific conformational changes in human prothrombin upon binding to procoagulant acidic lipid membranes. Thromb Haemost1994; 71(5): 596—604.
14.
Krishnaswamy S., Church WR, Nesheim ME, Mann KGActivation of human prothrombin by human prothrombinase. Influence of factor Va on the reaction mechanism. J Biol Chem1987; 262(7): 3291—3299.
15.
Puurunen M. , Vaarala O., Julkunen H., Aho K., Palosuo T.Antibodies to phospholipid-binding plasma proteins and occurrence of thrombosis in patients with systemic lupus erythematosus . Clin Immunol Immunopathol1996; 80(1): 16—22.
16.
Sorice M., Pittoni V., Circella A. et al. Anti-prothrombin but not “pure” anti-cardiolipin antibodies are associated with the clinical features of the antiphospholipid antibody syndrome. Thromb Haemost1998; 80(4): 713—715.
17.
Bertolaccini ML, Atsumi T., Khamashta MA, Amengual O., Hughes GRAutoantibodies to human prothrombin and clinical manifestations in 207 patients with systemic lupus erythematosus. J Rheumatol1998; 25(6): 1104—1108.
18.
Pengo V., Biasiolo A., Brocco T., Tonetto S., Ruffatti A.Autoantibodies to phospholipid-binding plasma proteins in patients with thrombosis and phospholipid-reactive antibodies. Thromb Haemost1996; 75(5): 721—724.
19.
Horbach DA, van Oort E., Donders RC, Derksen RH, de Groot PGLupus anticoagulant is the strongest risk factor for both venous and arterial thrombosis in patients with systemic lupus erythematosus. Comparison between different assays for the detection of antiphospholipid antibodies . Thromb Haemost1996; 76(6): 916—924.
20.
Forastiero RR, Martinuzzo ME, Cerrato GS, Kordich LC, Carreras LORelationship of anti beta2-glycoprotein I and anti prothrombin antibodies to thrombosis and pregnancy loss in patients with antiphospholipid antibodies. Thromb Haemost1997; 78(3): 1008—1014.
21.
Swadzba J., De Clerck LS, Stevens WJ et al. Anticardiolipin, anti-beta(2)-glycoprotein I, antiprothrombin antibodies, and lupus anticoagulant in patients with systemic lupus erythematosus with a history of thrombosis. J Rheumatol1997; 24(9): 1710—1715.
22.
Guerin J., Smith O., White B., Sweetman G., Feighery C., Jackson J.Antibodies to prothrombin in antiphospholipid syndrome and inflammatory disorders . Br J Haematol1998; 102(4): 896—902.
23.
Fleck RA, Rapaport SI, Rao LVAnti-prothrombin antibodies and the lupus anticoagulant. Blood1988; 72(2): 512—519.
24.
Arvieux J., Darnige L., Caron C., Reber G., Bensa JC, Colomb MGDevelopment of an ELISA for autoantibodies to prothrombin showing their prevalence in patients with lupus anticoagulants. Thromb Haemost1995; 74(4): 1120—1125.
25.
Galli M., Beretta G., Daldossi M., Bevers EM, Barbui T.Different anticoagulant and immunological properties of anti-prothrombin antibodies in patients with antiphospholipid antibodies. Thromb Haemost1997; 77(3): 486—491.
26.
Atsumi T., Ieko M., Bertolaccini ML et al. Association of autoantibodies against the phosphatidylserine-prothrombin complex with manifestations of the antiphospholipid syndrome and with the presence of lupus anticoagulant. Arthritis Rheum2000; 43(9): 1982—1993.
27.
Amengual O. , Atsumi T., Koike T.Specificities, properties, and clinical significance of antiprothrombin antibodies. Arthritis Rheum2003; 48(4): 886—895.
28.
Amengual O. , Atsumi T., Koike T.Antiprothrombin antibodies and the diagnosis of antiphospholipid syndrome. Clin Immunol2004; 112(2): 144—149.
29.
D'Agnillo P. , Levine JS, Subang R., Rauch J.Prothrombin binds to the surface of apoptotic, but not viable, cells and serves as a target of lupus anticoagulant autoantibodies . J Immunol2003; 170(6): 3408—3422.
30.
Rauch J., D'Agnillo P., Subang R., Levine JSAnti-phospholipid antibodies (aPL) and apoptosis: prothrombin-dependent aPL as a paradigm for phospholipid-dependent interactions with apoptotic cells . Thromb Res2004; 114(5—6): 371—382.
31.
Miyakis S., Lockshin MD, Atsumi T. et al. International consensus statement on an update of the classification criteria for definite antiphospholipid syndrome (APS). J Thromb Haemost2006; 4(2): 295—306.
32.
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 Immunol1995; 154(2): 954—960.
33.
Rao LV, Hoang AD, Rapaport SIMechanism and effects of the binding of lupus anticoagulant IgG and prothrombin to surface phospholipid. Blood1996; 88(11): 4173—4182.
34.
Akimoto T., Akama T., Kono I., Sumida T.Relationship between clinical features and binding domains of anti-prothrombin autoantibodies in patients with systemic lupus erythematosus and antiphospholipid syndrome. Lupus1999; 8(9): 761—766.
35.
Bevers EM, Galli M., Barbui T., Comfurius P., Zwaal RFLupus anticoagulant IgG's (LA) are not directed to phospholipids only, but to a complex of lipid-bound human prothrombin. Thromb Haemost1991; 66(6): 629—632.
36.
Oosting JD, Derksen RH, Bobbink IW, Hackeng TM, Bouma BN, de Groot PG.Antiphospholipid antibodies directed against a combination of phospholipids with prothrombin, protein C, or protein S: an explanation for their pathogenic mechanism?Blood1993; 81(10): 2618—2625.
37.
Field SL, Hogg PJ, Daly EB et al. Lupus anticoagulants form immune complexes with prothrombin and phospholipid that can augment thrombin production in flow. Blood1999; 94(10): 3421—3431.
38.
Field SL, Chesterman CN, Dai YP, Hogg PJLupus antibody bivalency is required to enhance prothrombin binding to phospholipid . J Immunol2001; 166(10): 6118—6125.
39.
Willems GM, Janssen MP, Comfurius P., Galli M., Zwaal RF, Bevers EMKinetics of prothrombin-mediated binding of lupus anticoagulant antibodies to phosphatidylserine-containing phospholipid membranes: an ellipsometric study. Biochemistry2002; 41(48): 14357—14363.
40.
Horbach DA, van Oort E., Derksen RH, de Groot PGThe contribution of anti-prothrombin-antibodies to lupus anticoagulant activity— discrimination between functional and non-functional anti-prothrombin-antibodies . Thromb Haemost1998; 79(4): 790—795.
41.
Zhao Y., Rumold R., Zhu M. et al. An IgG antiprothrombin antibody enhances prothrombin binding to damaged endothelial cells and shortens plasma coagulation times. Arthritis Rheum1999; 42(10): 2132—2138.
42.
Roubey RA, Pratt CW, Buyon JP, Winfield JBLupus anticoagulant activity of autoimmune antiphospholipid antibodies is dependent upon beta 2-glycoprotein I. J Clin Invest1992; 90(3): 1100—1104.
43.
Arnout J., Vanrusselt M., Wittevrongel C., Vermylen J.Monoclonal antibodies against beta-2-glycoprotein I: use as reference material for lupus anticoagulant tests. Thromb Haemost1998; 79(5): 955—958.
