重组猫白蛋白的特性分析
Characterization of recombinant cat albumin.
作者信息
Reininger R, Swoboda I, Bohle B, Hauswirth A W, Valent P, Rumpold H, Valenta R, Spitzauer S
机构信息
Institute of Medical and Chemical Laboratory Diagnostics, University of Vienna, Vienna, Austria.
出版信息
Clin Exp Allergy. 2003 Dec;33(12):1695-702. doi: 10.1111/j.1365-2222.2003.01817.x.
BACKGROUND
Indoor allergens derived from animals and mites often contribute to exacerbation of skin manifestations in atopic dermatitis (AD) patients.
OBJECTIVE
To produce and characterize recombinant cat albumin, a cross-reactive animal allergen.
METHODS
A complete cDNA coding for cat albumin was obtained by RT-PCR amplification from cat liver RNA. Recombinant cat albumin was expressed in Escherichia coli as hexahistidine-tagged protein, purified by nickel affinity chromatography and studied for IgE reactivity with sera from cat-allergic patients by ELISA and immunoblotting. Furthermore, CD203c expression of basophils from cat-allergic patients upon exposure to recombinant cat albumin was analysed.
RESULTS
Recombinant cat albumin, a cross-reactive animal allergen sharing most IgE epitopes with its natural counterpart, was produced in E. coli. It was recognized preferentially by IgE from AD patients and elicited IgE-dependent basophil activation in sensitized patients.
CONCLUSIONS
Recombinant cat albumin may be used as a paradigmatic tool to analyse mechanisms of allergen-triggered exacerbation of AD, for diagnostic and, perhaps for therapeutic purposes.
背景
源自动物和螨虫的室内过敏原常导致特应性皮炎（AD）患者皮肤症状加重。
目的
制备并鉴定重组猫白蛋白，一种交叉反应性动物过敏原。
方法
通过逆转录聚合酶链反应（RT-PCR）从猫肝脏RNA中扩增出编码猫白蛋白的完整cDNA。重组猫白蛋白在大肠杆菌中表达为带六聚组氨酸标签的蛋白，通过镍亲和层析纯化，并通过酶联免疫吸附测定（ELISA）和免疫印迹法研究其与猫过敏患者血清的IgE反应性。此外，分析了猫过敏患者的嗜碱性粒细胞在暴露于重组猫白蛋白后的CD203c表达。
结果
重组猫白蛋白是一种交叉反应性动物过敏原，与其天然对应物共享大多数IgE表位，在大肠杆菌中产生。它优先被AD患者的IgE识别，并在致敏患者中引发IgE依赖性嗜碱性粒细胞活化。
结论
重组猫白蛋白可作为一种典型工具，用于分析过敏原引发AD加重的机制，用于诊断，或许还可用于治疗目的。
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