title: The glycoprotein Ib-IX complex-specific monoclonal antibody SZ1 binds to a conformation-sensitive epitope on glycoprotein IX: implications for the target antigen of quinine/quinidine-dependent autoantibodies.
abstract: The monoclonal antibody SZ1 is of interest for two reasons: it was used to define complex formation between glycoprotein (GP) Ib and GP IX, and its epitope is likely to be identical to that recognized by most quinine- and quinidine-dependent autoantibodies that cause thrombocytopenia. To determine the location of the epitope for SZ1 within the GP Ib-IX complex (which consists of three subunits: GP Ib alpha, GP Ib beta, and GP IX), we tested the ability of the antibody to bind transfected cells that expressed different combinations of complex subunits, and compared this binding to the binding of antibodies of known specificity. SZ1 bound to cells that expressed the entire GP Ib-IX complex in the same pattern as did AN51 (an antibody specific for GP Ib alpha). However, unlike AN51, SZ1 did not bind alpha beta cells (ie, cells that express GP Ib alpha and GP Ib beta, but not GP IX), but did bind to beta IX and alpha IX cells. We then compared the binding patterns of SZ1 and FMC25, an antibody specific for GP IX. Both bound virtually identically to cell lines that expressed every combination of two of the three GP Ib-IX complex subunits. However, the epitopes of the two antibodies were not identical, because fixation with 4% paraformaldehyde of cells that expressed GP IX destroyed the SZ1 epitope while maintaining the FMC25 epitope. Because of the ability of SZ1 to block the binding of many quinine- and quinidine-dependent antibodies, these data strongly suggest that GP IX is the component of the GP Ib-IX complex recognized by those antibodies.
authors.affiliation: Gladstone Institute of Cardiovascular Disease, Department of Medicine, University of California, San Francisco.
authors.forename: J A
authors.initials: JA
authors.lastname: López
authors.forename: C Q
authors.initials: CQ
authors.lastname: Li
authors.forename: S
authors.initials: S
authors.lastname: Weisman
authors.forename: M
authors.initials: M
authors.lastname: Chambers
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology*; D001327:Autoimmune Diseases / Q000139:chemically induced / Q000276:immunology; D016466:CHO Cells; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D006224:Cricetinae; D000939:Epitopes / Q000187:drug effects / Q000276:immunology*; D005557:Formaldehyde / Q000494:pharmacology; D007739:L Cells; D051379:Mice; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000276:immunology*; D011108:Polymers / Q000494:pharmacology; D011487:Protein Conformation; D011802:Quinidine / Q000009:adverse effects*; D011803:Quinine / Q000009:adverse effects*; D013921:Thrombocytopenia / Q000139:chemically induced / Q000276:immunology; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D001324:Autoantigens; D000939:Epitopes; D010980:Platelet Membrane Glycoproteins; D011108:Polymers; D005557:Formaldehyde; D011803:Quinine; D011802:Quinidine; C003043:paraform