Expression, purification and characterization of Der f 27, a new allergen from dermatophagoides farinae.
Expression, purification and characterization of Der f 27, a new allergen from dermatophagoides farinae.
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发表时间：
2015-07
影响因子：
2.2
通讯作者：
Jianli Lin;Meng Li;Yulin Liu;Congli Jiang;Yulan Wu;Yuanyuan Wang;Anjian Gao;Zhigang Liu;
中科院分区：
文献类型：
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作者：
Jianli Lin;Meng Li;Yulin Liu;Congli Jiang;Yulan Wu;Yuanyuan Wang;Anjian Gao;Zhigang Liu;
屋尘螨(HDM)是一种重要的室内变应原来源，也是过敏性哮喘的主要诱因之一，其特性在尘螨变应原相关疾病的诊断和免疫治疗中具有重要意义。本研究旨在鉴定一种新的过敏原--粉尘螨衍生的丝胶蛋白(Der F 27)。本研究提取粉纹夜蛾的总RNA，克隆并表达了粉纹夜蛾Der f27基因。采用酶联免疫吸附试验、哮喘患者血清Western印迹和过敏者皮肤点刺试验(SPT)检测重组Der f 27蛋白的致敏性。建立r-Der-f27过敏性哮喘小鼠模型。克隆的Der f 27基因已在基因库中公布，登录号为KM009995。R-Der f 27阳性变态反应性疾病患者血清中r-Der f 27的IgE水平是正常人的3倍。19例DM-SPT阳性患者中，Der f27 SPT阳性率为42.1%。R-Der f 27致敏的过敏性哮喘小鼠的气道高反应性、血清特异性IgE和脾细胞培养上清液中的IL-4水平显著增加。综上所述，Der f 27是一种新的屋尘变应原亚型。
The house dust mite (HDM), Dermatophagoidesfarinae (D. farina), is one of the most important indoor allergen sources and a major elicitor of allergic asthma; itscharacterization is important in the diagnosis and immunotherapy of mite allergen-relevant diseases. This study aims to characterize a novel allergen, the D. farinae-derived serpin (Der f 27). In this study, the total RNA of D. farinae was extracted, and the Der f 27 gene was cloned and expressed. The allergenicity of recombinant Der f 27 protein was determined by enzyme-linked immunosorbent assay, and Western-blotting with the sera of asthma patients, and skin prick test (SPT) in allergic human subjects. A r-Der f 27 allergic asthma mouse model was established. The cloned Der f 27 gene has been presented at the Gene Bank with an accession number of KM009995. The IgE levels of r-Der f 27 in the serum from r-Der f 27 SPT positive allergic patients were 3 folds more than healthy subjects. The Der f 27 SPT positive ratewas 42.1% in 19 DM-SPT positive patients. Airway hyperresponsiveness, serum specific IgE, and levels of interleukin-4 in the spleen cell culture supernatant were significantly increased in allergic asthma mice sensitized to r-Der f 27. In conclusion, Der f 27 is a new subtype of house mite allergen.