Diagnosis and discrimination of autoimmune Graves' disease and Hashimoto's disease using thyroid-stimulating hormone receptor-containing recombinant proteoliposomes(MEDICAL BIOTECHNOLOGY)

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概要

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• Graves' disease (GD) is an autoimmune disease of the thyroid gland caused by autoantibodies against thyroid-stimulating hormone receptor (TSHR). Currently, the diagnostic test for TSHR autoantibodies is based on an indirect competitive binding assay that measures the ability of TSHR autoantibodies to inhibit the binding of thyroid-stimulating hormone (TSH) to TSHR. Here, we have developed a specific and direct diagnostic method for autoantibodies in GD that incorporates immobilized TSHR-containing recombinant proteoliposomes into an enzyme-linked immunosorbent assay (ELISA). To reduce non-specific binding of autoantibodies to recombinant proteoliposomes, we investigated the effect of polyethylene glycol (PEG)-lipid on the binding of commercially available anti-TSHR antibodies (aTSHRAb). The incorporation of PEG-lipids into liposomes decreased non-specific binding, as compared to liposomes that did not contain PEG-lipids, and the addition of blocking reagents further decreased non-specific reactivity. aTSHRAb exhibited higher reactivity towards PEG-modified TSHR recombinant proteoliposomes than PEG-modified liposomes without TSHR (bare liposomes). Importantly, serum autoantibodies from patients with GD, which is associated with hyperthyroidism, exhibited remarkably specific binding to TSHR recombinant proteoliposomes. Serum autoantibodies from patients with Hashimoto's disease (HD), which is associated with hypothyroidism, also reacted specifically with proteoliposomal TSHR. These results suggest that immobilized TSHR recombinant proteoliposomes can serve as a direct diagnostic test for GD and HD. Furthermore, given that there is no competition test currently available for detecting autoantibodies in HD, the combination of TSHR recombinant proteoliposome ELISA and indirect competitive TSHR binding assay might be an effective way to discriminate between GD and HD.

• 社団法人日本生物工学会の論文

著者

• Tsumoto Kanta

  Graduate School Of Engineering Mie University:liposome Engineering Laboratory Inc.

• Tsumoto Kanta

  三重大学 大学院生物資源学研究科

• Okumura Katsuzumi

  Graduate School Of Bioresources Mie University

• Okumura Katsuzumi

  三重大学 大学院生物資源学研究科

• IMAMURA Koji

  Medical and Biological Laboratories Co. Ltd

• Fukushima Hidetaka

  Graduate School of Bioresources, Mie University

• Matsuo Hideaki

  Liposome Engineering Laboratory, Inc.

• Morino Kazuhiko

  Medical and Biological Laboratories Co. Ltd.

• Yoshimura Tetsuro

  Graduate School of Engineering, Mie University

• Imamura Koji

  Medical And Biological Laboratories Co. Ltd.

• Matsuo Hideaki

  Liposome Engineering Laboratory Inc.

• Yoshimura Tetsuro

  Graduate School Of Engineering Mie University

• Yoshimura Tetsuro

  Graduate School Of Engineering Mie University:liposome Engineering Laboratory Inc.:nagoya Industrial

• Fukushima Hidetaka

  Graduate School Of Bioresources Mie University:graduate School Of Engineering Mie University

• Tsumoto Kanta

  Graduate School Of Engineering Mie University

関連論文

• 2P243 バキュロウイルス-リポソーム融合法によるカドヘリン組込みリポソームの構築と機能(生体膜・人工膜-構造・物性,第48回日本生物物理学会年会)
• Enzyme-linked immunosorbent assay using bacterial recombinant proteins of human BP230 as a diagnostic tool for bullous pemphigoid
• Diagnosis and discrimination of autoimmune Graves' disease and Hashimoto's disease using thyroid-stimulating hormone receptor-containing recombinant proteoliposomes(MEDICAL BIOTECHNOLOGY)
• Development of a Novel Preparation Method of Recombinant Proteoliposomes Using Baculovirus Gene Expression Systems
• 2P280 Asymmetric fusion between a giant unilamellar vesicle and large unilamellar vesicles applied to controlling a cell-sized microreactor(40. Membrane structure,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)
• 1P-185 コネキシン巨大プロテオリポソームの構築と機能(生体膜・人工膜-構造・物性,第47回日本生物物理学会年会)
• 2P281 Investigation of pH-induced membrane-fusion properties of baculovirus by microscopic observation of individual giant unilamellar vesicles(40. Membrane structure,Poster Session,Abstract,Meeting Program of EABS & BSJ 2006)

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