Development of diagnostic system for summer-type hypersensitivity pneumonitis

夏季型过敏性肺炎诊断系统的研制

• 批准号: 03557036
• 负责人: ANDO Masayuki
• 金额: $ 3.9万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03557036/
• 关键词: Japanese summer-type hypersensitivity pneumonitis; Trichosporon cutaneum; diagnostic system; enzyme-linked immunosorbent assay; glucuronoxylomannan; serotype-specific polysaccharide antigen; monoclonal antibody; 診断用ELISAキット; モノクロ-ナル抗体; 菌血清型関連抗原

Summer-type hypersensitivity pneumonitis is a unique type of hypersensitivity pneumonitis and the most prevalent in Japan. Our previous study clarified that the causative agent of the disease is Trichosporon cutaneum. In this study, we constructed a sandwich enzyme-linked immunosorbent assay system for diagnosis of summer-type hypersensitivity pneumonitis in which monoclonal antibody, D-8, specific for serotype II T.cutaneum was used to bind serotype-related polysaccharides to plastic plates, and this system was proven to have sufficient sensitivity and specificity.The affinity-purified antigen was shown to be essentially acidic polysaccharide comprising mannose, xylose, and glucuronic acid. Chemical analysis showed that this polysaccharide antigen contains a (1-3)-linked mannan backbone attached with short chains of (1-4)-linked mannose and a small portion of (1-2) linked xylose residues by substituting the 2- or 4- positions of the (1-3)-linked mannose residues of the main chain. The antigenic epitope was shown to involve the terminal glucuronic acid residues as revealed by immunodiffusion test and sandwich enzyme-linked immunosorbent assay using monoclonal antibody D-8.

夏季型超敏性肺炎是一种独特的超敏性肺炎，在日本最为流行。我们先前的研究明确了该疾病的病原体是皮癣毛丝虫病。在本研究中，我们构建了一个三明治型酶联免疫吸附检测系统用于诊断夏季型超敏性肺炎，该系统使用血清II型皮肤t细胞特异性单克隆抗体D-8将血清型相关多糖结合到塑料板上，并证明该系统具有足够的灵敏度和特异性。亲和纯化抗原被证明本质上是酸性多糖，包括甘露糖、木糖和葡萄糖醛酸。化学分析表明，该多糖抗原含有(1-3)-连接的甘露糖主链与(1-4)-连接的甘露糖短链相连，通过取代主链(1-3)-连接的甘露糖残基的2-或4-位置，含有一小部分（1-2）连接的木糖残基。单克隆抗体D-8的免疫扩散试验和夹心酶联免疫吸附试验表明，抗原表位涉及末端葡萄糖醛酸残基。

项目成果

Mizobe T, Yamasaki H, Doi K, Ando M, and Onoue K: "Analysis of serotype-specific antibodies to Trichosporon cutaneum Types U and II in patients with summer-type hypersensitivity pneumonitis with monoclonal antibodies to serotype-related polysaccharide ant

Mizobe T、Yamasaki H、Doi K、Ando M 和 Onoue K：“用针对血清型相关多糖蚂蚁的单克隆抗体分析夏季型过敏性肺炎患者中 U 型和 II 型皮肤毛孢子菌的血清型特异性抗体

Takanori Mizobe,Hisato Yamasaki,Kuniko Doi,Masayuki Ando.Kaoru Onoue: "Analysis of Serotype-specific Antibodies to Tricosporon cutaneum Types I and II in Patients with Summer-Type hyper sensitivity Pneumonitis with Monoclonal Antibodies to Serotype-Relate

Takanori Mizobe、Hisato Yamasaki、Kuniko Doi、Masayuki Ando.Kaoru Onoue：“用血清型相关单克隆抗体分析夏季型过敏性肺炎患者 I 型和 II 型皮肤三孢菌血清型特异性抗体