Biocenter Klein Flottbek and Botanical Garden, University of Hamburg, Hamburg, Germany.
Int Arch Allergy Immunol. 2012;159(3):235-42. doi: 10.1159/000337461. Epub 2012 Jun 20.
Over the past years, dust of green coffee beans has become known to be a relevant cause for occupational type I allergies. Up to now, allergy diagnostics is based on native green coffee bean extract which exhibits insufficient specificity due to interfering substances as well as batch-to-batch variations. No coffee allergen has been described on the molecular level so far. The aim of this study was to identify the first allergen of green coffee.
The allergenicity of native green coffee bean extracts was analyzed by means of ImmunoCAP in sera of 17 symptomatic coffee workers. A Coffea arabica pJuFo cDNA phage display library was constructed and screened for IgE binding to coffee proteins with 2 sera from allergic coffee workers. By sequence analysis, a new coffee allergen (Cof a 1) was identified, expressed in Escherichia coli, and evaluated by Western blots. The frequency of sensitization was investigated by ELISA screening.
The Cof a 1 cDNA encoded a 32-kDa C. arabica class III chitinase. Serum IgE antibodies to the recombinant allergen were found in 3 out of 17 symptomatic coffee workers (18%), whereas only 2 of them reacted to the commercial allergy test.
A class III chitinase of C. arabica was identified to be the first known coffee allergen Cof a 1. It may have a relevant potential for the specific diagnosis of coffee sensitization.
在过去的几年中,绿咖啡豆尘已被证明是职业 I 型过敏的一个相关原因。到目前为止,过敏诊断基于天然绿咖啡豆提取物,但由于干扰物质和批次间的变化,其特异性不足。到目前为止,还没有在分子水平上描述过任何咖啡过敏原。本研究的目的是鉴定绿咖啡的第一种过敏原。
通过免疫 CAP 分析了 17 名有症状的咖啡工人血清中天然绿咖啡豆提取物的致敏性。构建了阿拉比卡咖啡 pJuFo cDNA 噬菌体展示文库,并使用来自过敏咖啡工人的 2 种血清筛选与咖啡蛋白结合的 IgE。通过序列分析,鉴定了一种新的咖啡过敏原(Cof a 1),在大肠杆菌中表达,并通过 Western blot 进行评估。通过 ELISA 筛选调查了致敏的频率。
Cof a 1 cDNA 编码 32kDa 的阿拉伯咖啡 III 类几丁质酶。重组过敏原的血清 IgE 抗体在 17 名有症状的咖啡工人中的 3 名(18%)中发现,而其中只有 2 名对商业过敏测试有反应。
鉴定出的阿拉伯咖啡 III 类几丁质酶是已知的第一种咖啡过敏原 Cof a 1。它可能对咖啡致敏的特异性诊断具有相关的潜在作用。