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

许多植物过敏原已被发现，其过敏原研究领域呈现出更加复杂的阶段。过敏原之间的交叉反应经常被观察到。因此，复杂相将被称为“共同表位”。过敏原上的含asn表位被认为是常见的抗原之一。对大豆主要过敏原Gly - bd28k和小麦主要过敏原Tri - bd17k的糖段进行了分析。本文研究了小麦中主要过敏原Tria b27k的特性。首先，制备了针对该过敏原的单克隆抗体。采用双向电泳、等电电泳和SDS-PAGE分离小麦过敏原，并用上述抗体免疫印迹法检测凝胶上的蛋白。观察到对抗体有阳性反应的几个斑点。将凝胶上与正极相对应的区域切除并进行n端氨基酸测序。这些区域上的蛋白质被证明属于Tri a b27k。这一发现意味着Tri a b27k以多形体的形式出现。上述斑点与一种凝集素反应，该凝集素对虫草残留和小麦阳性患者的血清敏感。此外，切出对凝集素敏感的Tri a b27k并进行糖分析。随后检测甘露糖、n -乙酰氨基葡萄糖、半乳糖、呋喃糖和木糖的含量。有趣的是，在大肠杆菌中表达的重组Tri a b27k对小麦过敏患者的血清没有感染作用。本研究结果表明，上述患者血清中对过敏原敏感的IgE抗体大部分与过敏原的糖段结合。

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.