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Studies on allergens in fish and their processed products

鱼类及其加工品中过敏原的研究

基本信息

批准号：
11460097
负责人：
SHIOMI Kazuo
金额：
$ 4.42万
依托单位：
Tokyo University of Fisheries
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

项目摘要

1. Besides parvalbumin, the well-known fish allergen, a higher molecular weight allergen was newly detected in fish. The higher molecular weight allergen was purified from the bigeye tuna muscle and identified as collagen. Antigenic cross-reactivity was observed among fish collagens but not between collagens from fish and other animals such as mammals, crustaceans and mollusks.2. When 20 sera from fish-allergic patients were evaluated for their IgE reactivity to parvalbumin and collagen purified from the bigeye tuna muscle by ELISA, as many as 7 sera reacted to collagen. Furthermore, IgE binding ability of collagen was almost the same as that of parvalbumin and the content of collagen in muscle was comparable to that of parvalbumin. These results suggest that collagen is an important fish allergen.3. Heating of the homogenized fish muscle at 80ーC for 20 min was found to be most effective to simultaneously extract both parvalbumin and collagen. This extraction method is applicable to analysis of fish allergens by immunoblotting and diagnosis of fish allergy by RAST.4. The CDNA library was constructed from the mackerel muscle and CDNA encoding parvalbumin was cloned DNA sequence analysis revealed that the mackerel parvalbumin is composed of 108 amino acid residues, being a b-form. Elucidation of IgE-binding epitopes of the mackerel parvalbumin is under progress using various synthetic peptides covering the total amino acid sequence.5. Commercial fish meat paste products and their major raw material, surimi, were very low in allergenicity against parvalbumin-recognizing patients. In a model experiment using the walleye pollack muscle, parvalbumin was almost completely removed from the muscle during washing, suggesting that fish meat paste products made of fully washed surimi are hypoallergenic foods for parvalbumin-recognizing patients. The allergenicity (as to both parvalbumin and collagen) of fish extractives was also markedly reduced by digestion with proteases.

1.除了已知的鱼类过敏原小清蛋白外，在鱼类中新检测到一种更高分子量的过敏原。从大眼金枪鱼肌肉中分离纯化了高分子量的过敏原，并鉴定为胶原蛋白。鱼类胶原蛋白之间存在抗原交叉反应，但鱼类胶原蛋白与哺乳动物、甲壳类动物、软体动物等动物胶原蛋白之间不存在抗原交叉反应.当20名鱼类过敏患者的血清进行了评估，他们的IgE反应小白蛋白和胶原蛋白纯化的大眼金枪鱼肌肉ELISA，多达7血清反应胶原蛋白。此外，胶原蛋白的IgE结合能力与小清蛋白几乎相同，肌肉中胶原蛋白的含量与小清蛋白相当。这些结果表明胶原蛋白是一种重要的鱼类过敏原.在80 ° C下加热均质化的鱼肉20分钟被发现是同时提取小清蛋白和胶原蛋白的最有效方法。该提取方法适用于鱼类过敏原的免疫印迹分析和鱼类过敏原的RAST诊断.构建了鲭鱼肌肉cDNA文库，克隆了编码小清蛋白的cDNA，序列分析表明，鲭鱼小清蛋白由108个氨基酸组成，为b型。鲭鱼小清蛋白的IgE结合表位的解析正在进行中，使用覆盖总氨基酸序列的各种合成肽。市售鱼肉酱产品及其主要原料鱼糜对识别小白蛋白的患者的过敏性非常低。在一项使用青鳕肌肉的模型实验中，在清洗过程中，小清蛋白几乎完全从肌肉中去除，这表明由完全清洗的鱼糜制成的鱼肉糊产品对于识别小清蛋白的患者来说是低过敏性食物。鱼提取物的变应原性（如小清蛋白和胶原蛋白）也显着降低消化与蛋白酶。