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Thromb Haemost 1995; 74(04): 1120-1125
DOI: 10.1055/s-0038-1649891
Original Article
Coagulation
Schattauer GmbH Stuttgart

Development of an ELISA for Autoantibodies to Prothrombin Showing their Prevalence in Patients with Lupus Anticoagulants

J Arvieux
1   The Laboratoire d’lmnnunologie, Centre de Transfusion Sanguine, Grenoble, France
,
L Darnige
2   The Département de Biologie Clinique, CH Compiègne, France
,
C Caron
3   The Laboratoire d’Hématologie B, CHU Lille, France
,
G Reber
4   The Haemostasis unit, Geneva University Hospital, Switzerland
,
J C Bensa
1   The Laboratoire d’lmnnunologie, Centre de Transfusion Sanguine, Grenoble, France
,
M G Colomb
5   The CEA Laboratoire d’lmmunochimie, Inserm U238, DBMS, CEN-G, Grenoble, France
› Author Affiliations
Further Information

Publication History

Received 08 May 1995

Accepted after revision 19 June 1995

Publication Date:
09 July 2018 (online)

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Summary

Some lupus anticoagulants (LA) have been shown to be directed against phospholipid-bound prothrombin. While developing an ELISA to detect anti-prothrombin autoantibodies in patient serum or plasma, no or very low signal was observed using human prothrombin immobilized on plain polystyrene plates. In contrast, the same LA-positive samples bound specifically to prothrombin coated on γ-irradiated plates, depending on 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 ascertained by competition using liposome-bound prothrombin, since fluid-phase prothrombin competed poorly. Seventy-seven of 139 patients (55.4%) with LA related to a variety of underlying diseases possessed anti-prothrombin antibodies (27 IgG, 35 IgM and 15 both isotypes), either isolated or more often associated with anti-(β2 glycoprotein I (β2GPI) antibodies. These included 67-71% of the patients with systemic 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 shown for anti-β2GPI antibodies, IgG2 was the predominant IgG subclass reactive with prothrombin. Thus, autoimmune patients with LA have a high incidence of antibodies to β2GPI and prothrombin, the binding of which could similarly require high antigen density and/or exposure of cryptic epitopes resulting from protein interaction with an irradiated (i. e. more anionic) polystyrene surface.

 

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