Immunohistochemical Studies Using Immunized Guinea Pig Sera with Features of Anti-Human Thyroid, Eye and Skeletal Antibody and Graves’ Sera
Type 2 5′ deiodinase enzyme was observed in both thyroid and eye muscle tissues, highlighting its possible role as a common antigen in thyroid-associated ophthalmopathy. Sera of 105 Graves’ patients and 40 controls, and immunized guinea pig sera against TCSS peptide, showing homology to the amino acid sequence from 132 to 152 of type 2 5′ deiodinase, were investigated to demonstrate the binding effects to human thyroid, eye and skeletal muscle tissues.
Twenty-two Graves’ patients were positive for anti-TCSS peptide antibodies, of whom 18 cases had ophthalmopathy. The levels of anti-TCSS peptide antibodies were higher not only in Graves’ patients with (P<0.0001) and without (P<0.036) eye symptoms compared to controls but also the difference was significant between patients with and without ophthalmopathy (P<0.049). In Western blot, immunized sera showed binding reactions to the supernatant fractions of human thyroid, eye and skeletal muscle tissues at the range of 29 kDa. Patient sera with Graves’ ophthalmopathy resulted in positive reactions directed to membrane areas in thyroid follicular cells, and to fibers in eye and skeletal muscles using immunohistochemical method, while no positive staining was present after adding control sera. The binding features of immunized guinea pig sera exhibited similar staining in all human tissues but could be blocked with Graves’ sera. Our results suggest that type 2 5′ deiodinase enzyme protein could play a role as an antigen in Graves’ disease. Immunized guinea pig sera against TCSS peptide exhibited similar binding reactions and stainings to human thyroid, eye and skeletal muscle tissues as patient sera with Graves’ ophthalmopathy.
KEY WORDSType 2 5′ deiodinase Graves’ ophthalmopathy antibodies eye and skeletal muscle thyroid
- 7.Gunji K, De Bellis A, Kubota S, Swanson J, Wengrowicz S, Cochran B, Ackrell BAC, Salvi M, Bellastella A, Bizzarro A, Sinisi AA, Wall JR: Serum antibodies against the flavoprotein subunit of eye muscle autoimmunity in patients with Graves’ hyperthyroidism. J Clin Endocrinol Metab 84:1255–1262, 1999PubMedCrossRefGoogle Scholar
- 8.Gunji K, De Bellis A, Li AW, Yamada M, Kubota S, Ackrell B, Wengrowicz S, Bellastella A, Bizzarro A, Sinisi A, Wall JR: Cloning and characterization of the novel thyroid and eye muscle shared protein G2s: Autoantibodies against G2s are closely associated with ophthalmopathy in patients with Graves’ hyperthyroidism. J Clin Endocrinol Metab 85:1641–1647, 2000PubMedCrossRefGoogle Scholar
- 10.Gopinath B, Musselman R, Adams CL, Tani J, Beard N, Wall JR: Study of serum antibodies against three eye muscle antigens and the connective tissue antigen collagen XIII in patients with Graves’ disease with and without ophthalmopathy: Correlation with clinical features. Thyroid 16:967–974, 2006.PubMedCrossRefGoogle Scholar
- 24.Molnar I, Csathy L: Autoantibodies against the type 2 5′-deiodinase are involved in the development of Graves’ ophthalmopathy. In Program of 12th international Thyroid Congress, Kyoto, Japan, p 176 (Abstract), 2000Google Scholar
- 25.Nakahara R, Tsunekawa K, Yabe S, Nara M, Seki K, Kasahara T, Ogiwara T, Nishino M, Kobayashi I, Murakami M: Association of antipitiutary antibody and type 2 iodothyronine deiodinase antibody in patients with autoimmune thyroid disease. Endocrinol J 52:691–699, 2005Google Scholar