A radioligand binding assay to measure anti-thyroperoxidase autoantibodies in mice.
Autoimmune (Hashimoto's) thyroiditis is a chronic inflammatory disease which affects >3% of the population and shows an increasing prevalence with increasing age. Anti-thyroid autoantibodies, particularly against thyroperoxidase (also known as thyroid peroxidase or TPO), occur commonly in humans with autoimmune thyroid disease, and assays for anti-TPO autoantibodies are used in clinical diagnosis. In contrast anti-TPO autoantibodies have not been observed in classical mouse models of autoimmune thyroiditis, except in cases where mice were deliberately immunized with TPO. In the past, detection of anti-TPO autoantibodies in mice has relied on an indirect immunofluorescence assay (iIFA) which screens for thyroid follicle membrane staining in frozen sections of mouse thyroid glands. Since recent transgenic mouse models of autoimmune thyroiditis spontaneously develop anti-TPO autoantibodies, an assay other than serial dilution and iIFA would be useful to detect and quantify these autoantibodies. In this paper we describe such an assay, based on the capacity of autoimmune mouse sera to bind to the extracellular domain of mouse TPO which was produced in a radioactively labeled form using a coupled in vitro transcription/translation system. The same approach, using human TPO, could provide a highly sensitive method to detect anti-TPO autoantibodies in humans.
Hayward
Sarah L
SL
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada.
Suzuki
Kunimasa
K
Elliott
John F
JF
D000818:Animals; D000941:Antigens / Q000097:blood; D001323:Autoantibodies / Q000097:blood* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D003429:Cross Reactions; D004195:Disease Models, Animal; D019084:Fluorescent Antibody Technique, Indirect; D005968:Glutamate Decarboxylase / Q000235:genetics / Q000378:metabolism; D006683:HLA-DQ Antigens / Q000235:genetics; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics / Q000276:immunology*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D014176:Protein Biosynthesis; D011869:Radioligand Assay / Q000379:methods*; D013954:Thyroglobulin / Q000276:immunology; D013967:Thyroiditis, Autoimmune / Q000175:diagnosis*; D014158:Transcription, Genetic
D000941:Antigens; D001323:Autoantibodies; D006683:HLA-DQ Antigens; C096322:ICA512 autoantibody; D007527:Isoenzymes; D013954:Thyroglobulin; D007453:Iodide Peroxidase; D005968:Glutamate Decarboxylase; C401141:glutamate decarboxylase 2