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STUDY ON THE SUGAR MOIETY AS A COMMON ANTIGEN OF WHEAT ALLERGENS

小麦过敏原常见抗原糖部分的研究

基本信息

批准号：
15580112
负责人：
TSUJI Hideaki
金额：
$ 2.37万
依托单位：
Okayama Prefectural University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580112/
关键词：
Wheat allergen Tri a Bd 27K Tri a Bd 17K Sugar chain Allergenicity Common antigen Tri a Bd17K Tri a Bd 36K common epitope

项目摘要

Many plant allergens have been found, and show more complex phase concerning their allergen investigation field. Crossreaction between the allergens have been very often observed. Therefore, the complex phase would be unified by the idea entitle "a common epitope". Asn-containing epitopes on the allergens is considering as one of common antigens.We have elucidated the sugar moiety of soybean major allergen, Gly m Bd 28K and wheat major allergen Tri a Bd 17K.In the present study, the properties of Tria Bd 27K, a major allergen, in wheat, has been investigated in details. First of all, monoclonal antibodies against the allergen have been producwed. The allergen in wheat was separated by two-dimensional electrophoresis with isoelectric electrophoresis and SDS-PAGE, and the proteins on the gel were detected by immnoblotting with the above-mentioned antibodies. Several spots showing the positive response for the antibodies were observed. The regions on the gel corresponding to the positive spots were cut out and subjected to N-terminal amino acid sequenceing. The proteins on the regions were shown to belong to the Tri a Bd 27k. This finding means that Tri a Bd 27K occur as polyforms. The above-mentioned spots reacted with a lectin, sensitive to fukose residue and the sera of patients positive to wheat. Furthermore, the Tri a Bd 27K sensitive to the lectin were cut out and subjected to sugar analyses. Subsequently, mannose, N-acetylglucosamin were detected, and the occurrence of galactose and fukose and xylose were alsoshowed. Interestingly, a recombinant Tri a Bd 27K expressed in Eschericia coli was not shown to be insetive to the sera of patients with wheat allergy.The present findings demonstrate that most of IgE antibodies in the sera of the above patients, which were sensitive to the allergen, bind to the sugar moiety of the allergen.

植物过敏原种类繁多，其研究领域也呈现出较为复杂的局面。经常观察到过敏原之间的交叉反应。因此，复合物阶段将被称为“共同表位”的想法统一起来。本研究对大豆主要过敏原Gly m Bd 28 K和小麦主要过敏原Tri a Bd 17 K的糖基进行了鉴定，并对小麦主要过敏原Tri a Bd 27 K的特性进行了详细的研究。首先，针对过敏原的单克隆抗体已经产生。采用双向电泳结合等电电泳和SDS-PAGE技术分离小麦变应原，并以上述抗体为抗原进行免疫印迹检测。观察到几个显示抗体阳性反应的斑点。切下凝胶上对应于阳性斑点的区域，并进行N-末端氨基酸测序。这些区域上的蛋白质显示属于Tri a Bd 27 k。这一发现意味着Tri a Bd 27 K以多聚体形式存在。上述斑点与凝集素反应，该凝集素对小麦阳性患者的血清和真菌多糖残留敏感。此外，切下对凝集素敏感的Tri a Bd 27 K并进行糖分析。随后检测到甘露糖、N-乙酰氨基葡萄糖，并检测到半乳糖、岩藻糖和木糖的存在。有趣的是，在大肠杆菌中表达的重组Tri a Bd 27 K对小麦过敏患者的血清无免疫反应，本研究结果表明，上述患者血清中对过敏原敏感的IgE抗体大多与过敏原的糖部分结合。