title: Isolation of Epstein-Barr-virus-transformed lymphocytes producing IgG class monoclonal antibodies using a magnetic cell separator (MACS): preparation of thyroid-stimulating IgG antibodies from patients with Graves' disease.
abstract: In autoimmune diseases, IgG class autoantibodies are generally considered to be more pathognomonic than IgM class ones. Although Epstein-Barr virus (EBV)-transformation of lymphocytes is a useful method to obtain human monoclonal autoantibodies, it tends to result predominantly in IgM-producing cells. We depleted IgM+ cells before EBV-transformation with a Magnetic Cell Separator (MACS) in order to increase the chance of acquisition of cells producing IgG class anti-thyrotropin (TSH) receptor antibodies (TRAb). As a result, we obtained four independent B cell clones producing IgG class monoclonal thyroid-stimulating antibodies (TSAb) from three patients with Graves' disease. None of these clones showed any TSH binding inhibitor immunoglobulin (TBII) activity, suggesting independence of TSAb-producing lymphocytes from those producing TBII.
authors.affiliation: Department of Laboratory Medicine, Kyoto University, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.forename: T
authors.initials: T
authors.lastname: Akamizu
authors.forename: J
authors.initials: J
authors.lastname: Okuda
authors.forename: H
authors.initials: H
authors.lastname: Sugawa
authors.forename: F
authors.initials: F
authors.lastname: Matsuda
authors.forename: T
authors.initials: T
authors.lastname: Tsubata
authors.forename: T
authors.initials: T
authors.lastname: Mori
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D001710:Biotin; D002461:Cell Line, Transformed; D002999:Clone Cells / Q000276:immunology; D006111:Graves Disease / Q000276:immunology*; D004854:Herpesvirus 4, Human; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000276:immunology; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M / Q000276:immunology; D018828:Immunoglobulins, Thyroid-Stimulating / Q000096:biosynthesis*; D018189:Immunomagnetic Separation; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D018828:Immunoglobulins, Thyroid-Stimulating; D001710:Biotin