Humoral Autoimmunity in Gravs' Ophthalmopathy

Research Project

Project/Area Number	62570528
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	内分泌・代謝学
Research Institution	Kurume University
Principal Investigator	NONAKA Kyohei Kurume University, Professor, 医学部, 教授 (80028547)
Co-Investigator(Kenkyū-buntansha)	KUROKI Toshitaka Kurume University, Research fellow, 医学部, 助手 HIROMATSU Yuji Kurume University, Assistant Professor, 医学部, 講師 (10201740)
Project Period (FY)	1987 – 1989
Project Status	Completed (Fiscal Year 1989)
Budget Amount *help	¥1,900,000 (Direct Cost: ¥1,900,000) Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000) Fiscal Year 1987: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywords	Grves' Ophthalmopathy / Humoral Immunity / Western Blotting / Retroperitoneal Connective Tissue / Retroocular Connective Tissue / Tissue Crossreactivity / ウェスタンブロッテング / 抗外眼筋抗体 / CHAPSO / アフィニティーカラムクロマトグラフィー / enzyme linked immunosorbent assay / emzyme linked immunosorbent assay
Research Abstract	To test the hypothesis that humoral autoimmunity is involved as pathogenesis of Graves' ophthalmopathy(GO) we investigated an antigen antibody axis as yet unknown. Organs prepared for antigen were human retroorbital connective tissue, retroperitoneal connective tissue, extra ocular, muscle, thyroid, liver, spleen and heart. Antibodies were used as serum, crude IgG and affinity purified IgG using CHAPSO-solubilized retoperitoneal connective tissue membrane as antigen. Reaction of antibody with antigen was detected by SDS polyacrylamide gel electro- phoresis followed by Western blotting combined with immunodetection. 2 out of 15 cases with GO had an antibody against 54kDa membrane protein of retroperitoneal connective tissue. Whilst this antibody showed crossreactivity against membrane protein of retro orbital connective tissue and extraocular muscle, it's molecular weight was 29kDa. The reaction was very faint in the latter. 54kDa or 29kDa antigen did not exist in other organs tested. This antibody was not detected in 13 cases of 10 normals and I Hashimoto thyroiditis, I silent thyroiditis and I thyroid adenoma. We conclude I An immunologically identical antigenic determinant exist between retroperitoneal connective tissue, retroorbital connective tissue and extraocular muscle. 2 Membrane proteins on which this antigenic determinant exist differ in molecular weight, being 54kDa, 29kDa and 29kDa respectively. 3 Amount of 29kDa protein on extraocular muscle membrane is very little. 4 Other organs tested do not have these antigens. 5 Autoantibody against 54kDa protein is present in some of sera from patient with GO. Although significance of this antibody in pathogenesis of Go is not known at the present, this is a new potential antibody different from any antibodies ever found.