Molecular cloning of Per a 1 and definition of the cross-reactive Group 1 cockroach allergens
Molecular cloning of Per a 1 and definition of the cross-reactive Group 1 cockroach allergens
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DOI：
10.1016/s0091-6749(99)70430-6
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发表时间：
1999-05-01
影响因子：
14.2
通讯作者：
Chapman, MD
中科院分区：
文献类型：
--
作者：
Melén, E;Pomés, A;Chapman, MD
背景资料：对德国蟑螂和美国蟑螂产生的过敏原的致敏性与哮喘的病因密切相关，迄今为止鉴定的大多数蟑螂过敏原具有种属特异性。目的：本研究的目的是鉴定交叉反应的蟑螂过敏原并进行测序。用来自美国的对蟑螂过敏和致敏的患者的IgE抗体筛选美洲大蠊cDNA文库结果：获得了一个全长870 bp的cDNA克隆，开放阅读框为695 bp，编码231个氨基酸，分子量为26.2 kd。序列同源性搜索表明，Per a 1与5个先前报道的，但未鉴定的，来自B germanica和P americana的序列同源。这些序列编码具有多种分子大小的蛋白质，含有约100个氨基酸重复。Pera1序列也显示出与蚊子前体蛋白ANG12的31%同一性，ANG12可能参与消化。结论：本研究结果明确了美洲疟原虫和德国B虫产生的一个新的交叉反应性“第1组”变应原家族的分子结构和抗原关系。这些重组过敏原和特异性单克隆抗体将为蟑螂过敏性疾病的诊断和治疗提供新的思路。
Background: Sensitization to allergens produced by German and American cockroaches is strongly associated with the cause of asthma, Most of the cockroach allergens identified to date have been species specific.Objective: The aim of this study was to identify and sequence cross-reactive cockroach allergens.Methods: A Periplaneta americana cDNA library was screened with IgE antibody from patients in the United States who were allergic to cockroach and who were sensitized to Blattella germanica.Results: A cDNA clone was isolated that contained an 870-bp sequence with a 695-bp open reading frame, encoding a 231 amino acid protein, molecular weight 26.2 kd, Plaque immunoassays using anti-Bla gl and anti-Per a 1 mAbs and a panel of human IgE antibodies showed that the protein expressed by these clones was Per a 1. Sequence homology searches showed that Per a 1 was homologous to 5 previously reported, but unidentified, sequences from B germanica and P americana. These sequences encoded proteins with multiple molecular sizes containing approximately 100 amino acid repeats. The Per a 1 sequence also showed 31% identity to a mosquito precursor protein, ANG12, which may be involved in digestion. The Per a 1 cDNA was expressed in Pichia pastoris to produce purified recombinant allergen (yield, 14 mg/L).Conclusion: The results define the molecular structure and antigenic relationships between a new family of cross-reactive "Group 1" allergens produced by both P americana and B germanica. These recombinant allergens and specific mAbs will provide toots to improve the diagnosis and treatment of allergic diseases caused by cockroaches.