Direct method for detection and characterization of cell surface receptors for insulin by means of 125I-labeled autoantibodies against the insulin receptor.
Autoantibodies directed against the cell surface receptors for insulin are found in some patients with extreme insulin resistance. These antibodies specifically inhibit the binding of insulin to its receptor. A purified IgG fraction from one patient's plasma was labeled with 125I. The 125I-labeled antireceptor antibody, which initially represented about 0.3% of the total 125I-IgG, was enriched by selective adsorption and subsequent elution from cells rich in insulin receptors. The 125I-antireceptor antibody bound to cells and the binding was inhibited by whole plasma and purified IgG from this patient, as well as whole plasma from another patient with autoantibodies to the insulin receptor. Insulins that differed 300-fold in biological potency and affinity inhibited binding of 125I-antireceptor antibody in direct proportion to their ability to bind to the insulin receptor. The binding of 125I-antireceptor antibody was closely correlated with the binding of 125I-insulin over a wide range of receptor concentrations on different cell types. Experimentally induced reduction of the insulin receptor concentration was associated with parallel decreases in the binding of 125I-antireceptor antibody and 125I-insulin. The preparation of 125I-antireceptor antibody with a high specific activity by cytoadsorption and elution has provided a sensitive method for the detection of receptors and autoantibodies to cell surface components.
D000937:Antigen-Antibody Reactions; D001323:Autoantibodies; D001667:Binding, Competitive; D002478:Cells, Cultured; D003920:Diabetes Mellitus / Q000276:immunology; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007333:Insulin Resistance; D007700:Kinetics; D008214:Lymphocytes / Q000032:analysis*; D011972:Receptor, Insulin / Q000032:analysis* / Q000276:immunology
D001323:Autoantibodies; D007328:Insulin; D011972:Receptor, Insulin