Miyazawa H, Fukamachi H, Inagaki Y, Reese G, Daul C B, Lehrer S B, Inouye S, Sakaguchi M
Department of Medical Technology, Kyorin University, Tokyo, Japan.
J Allergy Clin Immunol. 1996 Nov;98(5 Pt 1):948-53. doi: 10.1016/s0091-6749(96)80011-x.
In Japan, squid is an important seafood, and some patients with food allergies are sensitive to squid. There has been no report, however, describing the major allergens of squid.
To characterize squid allergens, we isolated a major allergen from the Pacific flying squid (Todarodes pacificus) and compared it with a major allergen from a shrimp (Penaeus orientalis).
The major squid and shrimp allergens were isolated by column chromatography on diethylaminoethyl-Sepharose (Pharmacia, Uppsala, Sweden), hydroxylapatite, and Sephacryl S-300 (Pharmacia). The IgE reactivity of the isolated allergens was assessed by immunoblotting. The cross-reactivity between the squid and shrimp allergens was examined by use of mouse polyclonal and monoclonal antibodies to the major allergens. Amino acid sequence analyses of the isolated allergens were done.
The isolated squid allergen is a 38 kd, heat-stable protein. IgE antibody binding to the purified squid allergen was demonstrated by immunoblotting. Cross-reactivity between major squid and shrimp allergens was demonstrated with sera from patients allergic to squid or shrimp or with allergen-specific monoclonal antibodies. The amino acid sequence analysis of the major squid allergen showed a marked homology with tropomyosin from blood fluke planorbid (Biomphalaria glabrata), which is a common vector snail of Schistosoma mansoni.
This 38 kd protein is a major allergen of the squid, Todarodes pacificus, and is believed to be squid muscle protein tropomyosin. We named it Tod p 1 according to International Union of Immunological Societies allergen nomenclature regulation.
在日本,鱿鱼是一种重要的海产品,一些食物过敏患者对鱿鱼敏感。然而,尚无描述鱿鱼主要过敏原的报告。
为了鉴定鱿鱼过敏原,我们从太平洋褶柔鱼(Todarodes pacificus)中分离出一种主要过敏原,并将其与虾(东方对虾,Penaeus orientalis)的主要过敏原进行比较。
通过在二乙氨基乙基-琼脂糖凝胶(Pharmacia,瑞典乌普萨拉)、羟基磷灰石和Sephacryl S-300(Pharmacia)上进行柱色谱法分离鱿鱼和虾的主要过敏原。通过免疫印迹法评估分离出的过敏原的IgE反应性。使用针对主要过敏原的小鼠多克隆和单克隆抗体检测鱿鱼和虾过敏原之间的交叉反应性。对分离出的过敏原进行氨基酸序列分析。
分离出的鱿鱼过敏原是一种38 kd的热稳定蛋白。免疫印迹法证实了IgE抗体与纯化的鱿鱼过敏原结合。对鱿鱼或虾过敏患者的血清或过敏原特异性单克隆抗体证实了主要鱿鱼和虾过敏原之间的交叉反应性。主要鱿鱼过敏原的氨基酸序列分析显示与来自曼氏血吸虫常见中间宿主扁卷螺(Biomphalaria glabrata)的原肌球蛋白有显著同源性。
这种38 kd的蛋白是太平洋褶柔鱼的主要过敏原,被认为是鱿鱼肌肉蛋白原肌球蛋白。根据国际免疫学会联盟过敏原命名规则,我们将其命名为Tod p 1。