title: Inhibition of platelet GPIIb/IIIa binding to fibrinogen by serum factors: studies of circulating immune complexes and platelet antibodies in patients with hemophilia, immune thrombocytopenic purpura, human immunodeficiency virus-related immune thrombocytopenic purpura, and systemic lupus erythematosus.
abstract: We have studied the conditions of in vitro binding of platelet glycoprotein IIb/IIIa (GPIIb/IIIa) to fibrinogen and applied the results to identify and measure the serum inhibitors to the binding. For the enzyme-linked immunosorbent assay, platelet extract was delivered to a fibrinogen-coated microtiter plate that was incubated for 2 hours, followed by incubation with anti-GPIIb/IIIa monoclonal antibody for another 2 hours. The plate was then incubated with peroxidase-conjugated anti-mouse IgG for color development. The binding was shown to be calcium-dependent. The binding was partially blocked by treating the coated fibrinogen with anti-fibrinogen antibody. Reduction or dissociation of GPIIb/IIIa resulted in the total loss of its ability to bind to fibrinogen. Platelet extracts of patients with hemophilia showed decreased binding (25% and 14%, compared with control platelet extract), and an extract from a patient with Glanzmann's thrombasthenia showed no binding. With the enzyme-linked immunosorbent assay we have measured serum inhibitors to GPIIb/IIIa binding to fibrinogen in 35 hemophilia A, 17 immune thrombocytopenic purpura, 22 human immunodeficiency virus-related immune thrombocytopenic purpura, and 29 systemic lupus erythematosus serum samples. In those patients with inhibition by serum, polyethylene glycol precipitation of circulating immune complexes (CICs) decreased the inhibition by the supernatants, and all the resolubilized CIC precipitates demonstrated inhibition, which indicates that CICs play a major role in the inhibition of GPIIb/IIIa binding to fibrinogen. This, then, provides evidence of CIC-mediated impaired GPIIb/IIIa binding to fibrinogen in hemophilia A, HIV-ITP, and SLE.
authors.affiliation: Blood Research Institute, St. Michael's Medical Center, Newark, NJ 07102.
authors.forename: M
authors.initials: M
authors.lastname: Kamiyama
authors.forename: Y S
authors.initials: YS
authors.lastname: Arkel
authors.forename: K
authors.initials: K
authors.lastname: Chen
authors.forename: K
authors.initials: K
authors.lastname: Shido
mesh_terms: D000906:Antibodies / Q000378:metabolism; D000936:Antigen-Antibody Complex / Q000378:metabolism*; D001792:Blood Platelets / Q000276:immunology* / Q000378:metabolism / Q000648:ultrastructure; D001798:Blood Proteins / Q000502:physiology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000378:metabolism*; D006678:HIV / Q000302:isolation & purification; D006467:Hemophilia A / Q000097:blood*; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007519:Isoantigens / Q000378:metabolism*; D008180:Lupus Erythematosus, Systemic / Q000097:blood*; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011485:Protein Binding; D011696:Purpura, Thrombocytopenic / Q000097:blood* / Q000382:microbiology; D011961:Receptors, Fc / Q000302:isolation & purification
chemical_list: D000906:Antibodies; D000936:Antigen-Antibody Complex; D001798:Blood Proteins; D007074:Immunoglobulin G; D007519:Isoantigens; D010980:Platelet Membrane Glycoproteins; D011961:Receptors, Fc; D005340:Fibrinogen