Identification of Novel Allergenic Components from German Cockroach Fecal Extract by a Proteomic Approach
Identification of Novel Allergenic Components from German Cockroach Fecal Extract by a Proteomic Approach
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DOI：
10.1159/000347034
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发表时间：
2013-01-01
影响因子：
2.8
通讯作者：
Yong, Tai-Soon
中科院分区：
文献类型：
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作者：
Jeong, Kyoung Yong;Kim, Chung-ryul;Yong, Tai-Soon
背景：蟑螂产生强烈的过敏原，蟑螂粪便中的过敏原含量特别丰富。本研究分析了蟑螂粪便中的致敏成分，并对从粪便提取物中鉴定的重组α-淀粉酶的致敏性进行了评价。方法：采用蛋白质组学和免疫印迹技术对德国小蠊粪便提取物中的IgE反应蛋白进行分析。制备重组α-淀粉酶，ELISA法检测其致敏性。结果：分析德国小蠊粪便提取物鉴定出12种IgE反应性成分。这些过敏原中的大多数被发现是消化酶，如α-淀粉酶，胰蛋白酶，胰凝乳蛋白酶，金属蛋白酶，和中肠羧肽酶A，但身份的3 IgE反应蛋白仍然是未知的。大豆球蛋白样蛋白，这可能是来自蟑螂的饮食，也被确定。德国小蠊α-淀粉酶与猪α-淀粉酶的同源性最高（55.8%），其次是螨群4变应原（Blo t4，50.4%; Der p4，49.8%; Eur m4，47.4%）。本研究表达了重组德国小蠊α-淀粉酶，并用ELISA法检测了其变应原性。在德国小蠊致敏的受试者中，41.4%（12/29）的血清样品中检测到抗重组淀粉酶的特异性IgE。重组α-淀粉酶能够抑制德国小蠊全身提取物55%的特异性IgE。结论：淀粉酶是蟑螂粪便中一种重要的新变应原。希望重组α-淀粉酶能为进一步的研究和临床应用奠定基础。版权所有（C）2013 S. Karger AG，巴塞尔
Background: Cockroaches produce potent allergens, and cockroach feces are known to be especially rich in allergens. In this study, we analyze the allergenic components from cockroach feces and evaluate allergenicity of recombinant a-amylase identified from fecal extract. Methods: IgE-reactive proteins from German cockroach fecal extract were analyzed by proteomic analysis and immunoblotting. Recombinant a-amylase was produced and its allergenicity was evaluated by ELISA. Results: Analysis of German cockroach fecal extracts identified 12 IgE-reactive components. Most of these allergens were found to be digestive enzymes such as a-amylase, trypsin, chynnotrypsin, metalloprotease, and midgut carboxypeptidase A, but the identity of 3 IgE-reactive proteins is still unknown. Glycinin-like proteins, which were likely derived from the cockroach diet, were also identified. German cockroach a-amylase shares the highest identity with pig alpha-amylase (55.8%), followed by mite group 4 allergens (Blo t 4, 50.4%; Der p 4, 49.8%; Eur m 4, 47.4%). In this study, recombinant a-amylase from German cockroach was expressed, and its allergenicity was examined by ELISA. Specific IgE against recombinant amylase was detected in 41.4% (12/29) of serum samples from German cockroach-sensitized subjects. Recombinant a-amylase was able to inhibit 55% of specific IgE to German cockroach whole-body extract. Conclusions: Amylase was found to be an important novel allergen in cockroach feces. It is hoped that recombinant a-amylase will be useful for further studies and clinical applications. Copyright (C) 2013 S. Karger AG, Basel