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.
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.
Li
H
H
Department of Laboratory Medicine, Kyoto University, Japan.
Akamizu
T
T
Okuda
J
J
Sugawa
H
H
Matsuda
F
F
Tsubata
T
T
Mori
T
T
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
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D018828:Immunoglobulins, Thyroid-Stimulating; D001710:Biotin