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皮屑芽生菌II型血清型特异性多糖抗原的纯化与鉴定:日本夏季型过敏性肺炎的一种疾病相关抗原

Purification and characterization of the serotype-specific polysaccharide antigen of Trichosporon cutaneum serotype II: a disease-related antigen of Japanese summer-type hypersensitivity pneumonitis.

作者信息

Mizobe T, Ando M, Yamasaki H, Onoue K, Misaki A

机构信息

Department of Immunology, Kumamoto University School of Medicine, Japan.

出版信息

Clin Exp Allergy. 1995 Mar;25(3):265-72. doi: 10.1111/j.1365-2222.1995.tb01039.x.

Abstract

Summer-type hypersensitivity pneumonitis (SHP) 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, and that the patients with SHP have high titres of antibodies against the serotype-specific antigen of polysaccharide nature which exist in the high molecular weight fraction of the culture supernatant of the yeast. In this study, we purified the serotype-specific antigen of serotype II T. cutaneum by gel filtration and affinity chromatography using a monoclonal antibody, D-8, specific for a high molecular weight antigen of serotype II T. cutaneum, and elucidated the structure of the antigen. This affinity-purified antigen was shown to be an essentially acidic polysaccharide comprising mannose, xylose, and glucuronic acid (6:44:4.7). Chemical analysis showed that this polysaccharide antigen contains a (1-3)-linked mannan backbone attached with short side chains of (1-4)-linked mannose and a small proportion of (1-2)-linked xylose residues by substituting the 2- or 4-positions of the (1-3)-linked mannose residues of the main chain. Approximately one-fifth of the side chains were terminated with glucuronic acid residues. The antigenic epitope of the serotype-specific antigen was shown to involve the terminal glucoronic acid residues as revealed by immunodiffusion test and sandwich enzyme-linked immunosorbent assay using monoclonal antibody D-8.

摘要

夏季型过敏性肺炎(SHP)是一种独特类型的过敏性肺炎,在日本最为常见。我们之前的研究阐明,该疾病的病原体是皮状丝孢酵母,并且SHP患者针对酵母培养上清液高分子量部分中存在的多糖性质的血清型特异性抗原具有高滴度抗体。在本研究中,我们使用针对皮状丝孢酵母II型高分子量抗原的单克隆抗体D-8,通过凝胶过滤和亲和层析纯化了皮状丝孢酵母II型的血清型特异性抗原,并阐明了该抗原的结构。这种亲和纯化的抗原被证明是一种基本为酸性的多糖,由甘露糖、木糖和葡萄糖醛酸组成(比例为6:44:4.7)。化学分析表明,这种多糖抗原含有一个(1-3)连接的甘露聚糖主链,其连接着(1-4)连接的甘露糖短侧链以及一小部分(1-2)连接的木糖残基,这些木糖残基通过取代主链中(1-3)连接的甘露糖残基的2或4位而连接。大约五分之一的侧链以葡萄糖醛酸残基结尾。免疫扩散试验和使用单克隆抗体D-8的夹心酶联免疫吸附测定表明,血清型特异性抗原的抗原表位涉及末端葡萄糖醛酸残基。

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