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从悬铃木花粉中表达、纯化和分析 Pla a 3 过敏原。

Expression, purification and epitope analysis of Pla a 3 allergen from Platanus acerifolia pollen.

机构信息

Research Division of Clinical Pharmacology, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210029, P.R. China.

Department of Allergy, Peking Union Medical College Hospital, Peking Union Medical College, Chinese Academy of Medical Sciences, Beijing 100730, P.R. China.

出版信息

Mol Med Rep. 2017 Sep;16(3):2851-2855. doi: 10.3892/mmr.2017.6867. Epub 2017 Jun 28.

Abstract

Platanus acerifolia (P. acerifolia) is an important cause of pollinosis in cities. The use of allergen extracts on patients with allergic diseases is the most commonly applied method to attempt to treat pollinosis. Pla a 3, a non‑specific lipid transfer protein, is a major allergen present in P. acerifolia pollen extracts. In the present study, the Pla a 3 gene was sub‑cloned into a pSUMO‑Mut vector using Stu I and Xho I sites and transformed into the Arctic Express™ (DE3) RP E. coli host strain. The purified Pla a 3 allergen was analyzed by western blotting and the results revealed that the Pla a 3 allergen has the ability to bind IgE in the P. acerifolia pollen of allergic patients' sera. Moreover, the authors predicted the potential B cell epitopes of the Pla a 3 allergen using the DNAStar Protean system, the Bioinformatics Predicted Antigenic Peptides system and the BepiPred 1.0 server. In addition, the T cell epitopes were predicted by the SYFPEITHI database and the NetMHCII‑2.2 server. As a result, two B cell epitopes (35‑45 and 81‑86) and four potential T cell epitopes including 2‑15, 45‑50, 55‑61 and 67‑73 were predicted in the present study. The current results can be used to contribute to allergen immunotherapies and useful in peptide‑based vaccine designs of pollen allergy.

摘要

悬铃木(P. acerifolia)是城市花粉症的重要致病原。在过敏性疾病患者中使用过敏原提取物是尝试治疗花粉症最常用的方法。Pla a 3,一种非特异性脂质转移蛋白,是悬铃木花粉提取物中的主要过敏原。在本研究中,使用 Stu I 和 Xho I 位点将 Pla a 3 基因亚克隆到 pSUMO-Mut 载体中,并转化到 Arctic Express™(DE3)RP E. coli 宿主菌株中。通过 Western blot 分析纯化的 Pla a 3 过敏原,结果表明 Pla a 3 过敏原能够与过敏患者血清中的悬铃木花粉中的 IgE 结合。此外,作者使用 DNAStar Protean 系统、Bioinformatics Predicted Antigenic Peptides 系统和 BepiPred 1.0 服务器预测了 Pla a 3 过敏原的潜在 B 细胞表位。此外,通过 SYFPEITHI 数据库和 NetMHCII-2.2 服务器预测了 T 细胞表位。结果,在本研究中预测了两个 B 细胞表位(35-45 和 81-86)和四个潜在的 T 细胞表位,包括 2-15、45-50、55-61 和 67-73。目前的结果可用于过敏原免疫治疗,并有助于花粉过敏的基于肽的疫苗设计。

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