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Detection of Autoantibodies to Thyroid Peroxidase in Autoimmune Thyroid Diseases by Micro-ELISA and Immunoblotting^*

Central Laboratory of Clinical Investigation, Miyazaki Medical College Hospital Kiyotake, Miyazaki 889-16:
Department of Internal Medicine, Miyazaki Prefectural Hospital Miyazaki 880, Japan

Address all correspondence and requests for reprints to: Sachiya Ohtaki, M.D., Central Laboratory of Clinical Investigation, Miyazaki Medical College Hospital, Kiyotake, Miyazaki 889-16, Japan.

Serum autoantibodies to thyroid peroxidase (TPO) in patients with thyroid autoimmune diseases were studied by micro-ELISA and immunoblotting. Twenty-four patients, 15 with Graves' disease and 9 with Hashimoto's thyroiditis, whose serum titers were greater than 3200 on the microsomal hemagglutination test (except for 1 patient with a titer of 800) had autoantibodies to TPO. Both immunoglobulin G and M classes of autoantibodies were detected, with the former being more prominent. When TPO and thyroid microsomes were used as a target in a competitive binding inhibition test, the results suggested that TPO was a major thyroid microsomal antigen. On the other hand, immunoblotting analysis showed 3–4 bands in the 45–60K region stained by patients' sera in addition to human TPO with mol wt of 100K and 107K; only the latter 2 bands stained with antiporcine TPO antibody. In the majority of sera, TPO bands were clearer than others, although some sera showed the clearest band with a mol wt of 55K. These results indicate that patients with autoimmune thyroid disease often have autoantibodies to TPO that can be detected by micro- ELISA and immunoblotting, and that TPO is a major component of the thyroid microsomal antigen.

^* This work was supported in part by a Special Grant for Clinical Research and Grants-in-Aid for Scientific Research from the Ministry of Education, Science, and Culture of Japan (no. 60480274).

Received September 24, 1985.

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