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

• 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

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.

许多植物过敏原已被发现，并且其过敏原研究领域呈现出更加复杂的阶段。经常观察到过敏原之间的交叉反应。因此，复杂的相将由名为“共同表位”的想法统一。过敏原上含有 Asn 的表位被认为是常见抗原之一。我们已经阐明了大豆主要过敏原 Gly m Bd 28K 和小麦主要过敏原 Tria Bd 17K 的糖部分。在本研究中，Tria Bd 27K 的特性小麦中的一种主要过敏原已被详细研究。首先，针对过敏原的单克隆抗体已经生产出来。通过等电电泳和SDS-PAGE二维电泳分离小麦中的过敏原，并用上述抗体进行免疫印迹检测凝胶上的蛋白质。观察到几个显示抗体阳性反应的点。切下凝胶上对应于阳性点的区域并进行N端氨基酸测序。这些区域上的蛋白质被证明属于 Tri a Bd 27k。这一发现意味着 Tri a Bd 27K 以多晶型形式出现。上述斑点与凝集素发生反应，该凝集素对岩藻糖残留物和小麦呈阳性的患者血清敏感。此外，对凝集素敏感的Tri a Bd 27K被切出并进行糖分析。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 27K并未显示出对小麦过敏患者的血清无作用。目前的研究结果表明，上述患者血清中的大多数IgE抗体对过敏原敏感，与过敏原的糖部分结合。

项目成果

Identification of the 23-kDa peptide derived from th precursor of Gly m Bd 28K, a major soybean allergen, as a new allergen.

将源自主要大豆过敏原 Gly m Bd 28K 前体的 23-kDa 肽鉴定为新过敏原。

• 发表时间: 2004
• 期刊: Biochim.Biophys.Acta. 1673(1-2)
• 作者: M.Hiemori;et al.
• 通讯作者: et al.