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Antibodies to Human Thyroid Peroxidase in Autoimmune Thyroid Disease: Studies with a Cloned Recombinant Complementary Deoxyribonucleic Acid Epitope^*

Université Libre de Bruxelles, IRIBHN Brussels, Belgium
Cattedra di Endocrinologia, Uniuersita di Pisa (S.M., P.P., F.S., A.P.) Pisa, Italy
Laboratoire de Biochimie Médicale, Université de Marseille (J.R.) Marseille, France

Address all correspondence and requests for reprints to: Marian Ludgate, Ph.D., Université Libre de Bruxelles, Institut de Recherche Interdisciplinaire en Biologie Humaine et Nucleaire. Faculté de Médecine, Campus Erasme, Route de Lennik 808,1070 Brussels, Belgique.

Previous studies carried out by screening a gtll human thyroid cDNA library with serum samples from selected patients with Hashimoto's thyroiditis and a polyclonal antibody to porcine thyroid peroxidase (TPO) confirmed, at the molecular level, that TPO is a major component of the thyroid microsomal antigen (M). That investigation led to the isolation of a clone (C2) which encodes an 85-amino acid segment of TPO and harbors a major epitope recognized by serum from several patients with autoimmune thyroid disease that contained anti-M autoantibodies (MAb). In this study, C2 antigen that was produced as a β-galactosidase fusion protein was used to establish an enzyme-linked immunoabsorbent assay for the detection of anti-C2 autoantibodies (C2Ab). C2Ab then were assayed in 191 patients with different autoimmune and nonautoimmune thyroid disorders, and 50 patients with nonthyroidal autoimmune diseases. The results were compared with the titers of anti-TPO antibodies (TPOAb; as detected by monoclonal antibody- assisted RIA) and MAb (as detected by passive hemagglutination). Positive C2Ab was found in the serum of 85 of 136 (63%) patients whose serum contained TPOAb and/or MAb. A significant positive correlation was found between the levels of C2Ab and those of TPOAb (r = 0.76; P < 0.001) or MAb (r = 0.69; P < 0.001), which was independent of the type of underlying autoimmune thyroid disorder. Low levels of C2Ab also were found in 10 of 105 (9%) serum samples that did not contain TPOAb. Western blot analysis carried out on the latter samples showed that in 2 samples the apparent C2Ab reactivity was due to the presence of antibodies reacting with β-galactosidase.

In conclusion, we confirmed the validity of screening gtll cDNA human thyroid libraries to better characterize thyroid autoantigens and demonstrated the feasibility of using recombinant proteins to establish diagnostic assays for autoantibodies.

^* This work was supported in part by Techland S.A. Belgium and the Ministère des Affaires Wallonnes, the National Research Council (CNR, Rome, Italy), Target Project Preventive Medicine and Rehabilitation, Subproject Mechanisms of Aging (Grant 87.00403.556), and Ente Nazionale per la Ricerca e lo Sviluppo della Energia Nucleare e delle Energie Alternative (E.N.E.A., Rome, Italy; to A.P.).

Received September 13, 1988.

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