DEVELOPMENT OF AN ELISA FOR AUTOANTIBODIES TO PROTHROMBIN SHOWING THEIR PREVALENCE IN PATIENTS WITH LUPUS ANTICOAGULANTS

Citation
J. Arvieux et al., DEVELOPMENT OF AN ELISA FOR AUTOANTIBODIES TO PROTHROMBIN SHOWING THEIR PREVALENCE IN PATIENTS WITH LUPUS ANTICOAGULANTS, Thrombosis and haemostasis, 74(4), 1995, pp. 1120-1125
Citations number
28
Categorie Soggetti
Hematology,"Cardiac & Cardiovascular System","Peripheal Vascular Diseas
Journal title
ISSN journal
03406245
Volume
74
Issue
4
Year of publication
1995
Pages
1120 - 1125
Database
ISI
SICI code
0340-6245(1995)74:4<1120:DOAEFA>2.0.ZU;2-9
Abstract
Some lupus anticoagulants (LA) have been shown to be directed against phospholipid-bound prothrombin. While developing an ELISA to detect an ti-prothrombin autoantibodies in patient serum or plasma, no or very l ow signal was observed using human prothrombin immobilized on plain po lystyrene plates. In contrast, the same LA-positive samples bound spec ifically to prothrombin coated on gamma-irradiated plates, depending o n the radiation dose, in the absence of added calcium and phospholipid . Optimization of the assay required the addition of 0.1% Tween 20 to the buffers. Antibody specificity for immobilized prothrombin was asce rtained by competition using liposome-bound prothrombin, since fluid-p hase prothrombin competed poorly. Seventy-seven of 139 patients (55.4% ) with LA related to a variety of underlying diseases possessed anti-p rothrombin antibodies (27 IgG, 35 IgM and 15 both isotypes), either is olated or more often associated with anti-beta(2) glycoprotein I (beta (2)GPI) antibodies. These included 67-71% of the patients with systemi c lupus erythematosus and related disorders, primary antiphospholipid antibody syndrome or drug-induced LA (autoimmune groups), but only 19- 20% of those with infection or malignancy (p <0.001). As previously sh own for anti beta(2)GPI antibodies, IgG(2) was the predominant IgG sub class reactive with prothrombin. Thus, autoimmune patients with LA. ha ve a high incidence of antibodies to beta(2)GPI and prothrombin, the b inding of which could similarly require high antigen density and/or ex posure of cryptic epitopes resulting from protein interaction with an irradiated (i. e. more anionic) polystyrene surface.