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Thromb Haemost 1996; 76(06): 1020-1029
DOI: 10.1055/s-0038-1650702
Original Article
Schattauer GmbH Stuttgart

PAICA: A Method for Characterizing Platelet-Associated Antibodies - Its Application to the Study of Idiopathic Thrombocytopenic Purpura and to the Detection of Platelet-bound c7E3

Laurent Macchi
The UMR 5533 CNRS, Institut Fédératif “Coeur-Vaisseaux-Thrombose”, Hôpital Cardiologique, Pessac, France
,
Gisèle Clofent-Sanchez
The UMR 5533 CNRS, Institut Fédératif “Coeur-Vaisseaux-Thrombose”, Hôpital Cardiologique, Pessac, France
,
Gérald Marit
1   Service des Maladies du Sang, Centre François Magendie, Hôpital du Haut-Lévèque, Pessac, France
,
Claude Bihour
The UMR 5533 CNRS, Institut Fédératif “Coeur-Vaisseaux-Thrombose”, Hôpital Cardiologique, Pessac, France
,
Catherine Durrieu-Jais
2   Unité de Soins Intensifs, Hôpital Cardiologique, Pessac, France
,
Pierre Besse
2   Unité de Soins Intensifs, Hôpital Cardiologique, Pessac, France
,
Paquita Nurden
The UMR 5533 CNRS, Institut Fédératif “Coeur-Vaisseaux-Thrombose”, Hôpital Cardiologique, Pessac, France
,
Alan T Nurden
The UMR 5533 CNRS, Institut Fédératif “Coeur-Vaisseaux-Thrombose”, Hôpital Cardiologique, Pessac, France
› Author Affiliations
Further Information

Publication History

Received 08 January 1996

Accepted after resubmission 20 August 1996

Publication Date:
11 July 2018 (online)

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Summary

In idiopathic thrombocytopenic purpura (ITP), autoantibodies reacting with antigens on the platelet membrane bring about accelerated platelet destruction. We now report PAICA (“Platelet-Associated IgG Characterization Assay”), a method for detecting autoantibodies bound to specific membrane glycoproteins in total platelet lysates. This monoclonal antibody (MAb) capture assay takes into account the fact that antibodies on circulating platelets may be translocated to internal pools as well as being on the surface. A total of twenty ITP patients were examined by PAICA, and the results compared with those obtained by measuring (i) serum antibodies bound to paraformaldehyde-fixed control platelets by ELISA, (ii) IgG bound to the surface of the patient’s own platelets by flow cytometry (PSIgG), (iii) total platelet-associated IgG (PAIgG) by ELISA and (iv) serum antibodies reacting with control platelets by MAIPA (“Monoclonal Antibody-specific Immobilization of Platelet Antigens”). Of twelve patients with elevated PAIgG, nine had increased PSIgG yet eleven reacted positively in PAICA. Of these, eight possessed antibodies directed against GP Ilb-IIIa, two against GP Ib-IX and one patient possessed antibodies directed against GP Ilb-IIIa and GP Ia-IIa respectively. Only seven of the patients possessed serum antibodies detectable by MAIPA. PAICA was also able to detect platelet-associated c7E3 (the chimeric form of Fab fragments of the MAb 7E3) following its infusion during antithrombotic therapy, when it proved more sensitive over a seven-day period than a MAIPA assay adapted for assessing surface-bound antibody. We propose that PAICA provides added sensitivity to the detection of platelet-associated antibodies in immune thrombocytopenias or following therapy with humanized MAbs.

 

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