鉴定Der p 39（一种新型过敏原）的免疫显性IgE表位 。 注：原文中“a novel allergen of.”后面似乎缺少内容。
Identification of an immunodominant IgE epitope of Der p 39, a novel allergen of .
作者信息
Li Wei-Yong, Cai Ze-Lang, Zhang Bo-Ping, Chen Jia-Jie, Ji Kunmei
机构信息
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Health Science Center, Shenzhen University, No. 1066 Xueyuan Road, Nanshan District, Shenzhen 518060, China.
出版信息
World Allergy Organ J. 2022 May 6;15(5):100651. doi: 10.1016/j.waojou.2022.100651. eCollection 2022 May.
BACKGROUND
House dust mites (HDMs) are the main source of indoor inhalatory allergens that cause IgE-mediated allergic diseases. The discovery and identification of HDM allergens are important for the diagnosis and treatment of allergic diseases.
OBJECTIVE
We sought to identify a Group 39 (Der p) allergen, namely Der p 39, and explore its immunodominant IgE epitopes.
METHODS
Homology analysis of amino acid (aa) sequences in HDM and human troponin C (TnC)-like protein was performed. Total RNA of Der p was extracted and used to amplify Der p 39 cDNA with specific primers. Recombinant Der p 39 protein was expressed with a pET-His prokaryotic expression system and purified with Ni-NTA resins. IgE binding was evaluated with western blot, dot blot, and enzyme-linked immunosorbent assay (ELISA) experiments. The IgE binding epitopes of Der p 39 were identified by observing HDM-allergic sera interactions with truncated and hybrid proteins formed from Der p 39 and human TnC-like proteins.
RESULTS
The Der p 39 open reading frame (ORF) cDNA was found to be 462 base pairs and registered in the NCBI library (GenBank no. MZ336019.1). Der p 39, which encoded 153 aa, was found to have 35.63% and 99.35% homology with human TnC and (Der f) 39, respectively. IgE-ELISA showed IgE binding with expressed and purified recombinant Der p 39 (18 kDa) in 5/87 (5.75%) HDM-allergic sera samples. Analyses of IgE binding with Der p 39-based truncated and hybrid proteins indicated that IgE binding epitopes are likely located in the C-terminal region and dependent on conformational structure. The data from this study were submitted to the World Health Organization and International Union of Immunological Societies (WHO/IUIS) Allergen Nomenclature database.
CONCLUSION
Der p 39 was identified as a minor HDM allergen with a conformational IgE binding epitope. These findings could have important theoretical implications in the development of HDM allergy diagnostics and therapeutics.
背景
屋尘螨（HDM）是引起IgE介导的过敏性疾病的室内吸入性过敏原的主要来源。HDM过敏原的发现和鉴定对于过敏性疾病的诊断和治疗具有重要意义。
目的
我们试图鉴定一种39组（Der p）过敏原，即Der p 39，并探索其免疫显性IgE表位。
方法
对HDM和人肌钙蛋白C（TnC）样蛋白的氨基酸（aa）序列进行同源性分析。提取Der p的总RNA，并用特异性引物扩增Der p 39 cDNA。用pET-His原核表达系统表达重组Der p 39蛋白，并用Ni-NTA树脂纯化。通过蛋白质印迹、斑点印迹和酶联免疫吸附测定（ELISA）实验评估IgE结合情况。通过观察HDM过敏血清与由Der p 39和人TnC样蛋白形成的截短蛋白及杂交蛋白的相互作用，鉴定Der p 39的IgE结合表位。
结果
发现Der p 39开放阅读框（ORF）cDNA为462个碱基对，并已在NCBI文库中登记（GenBank编号：MZ336019.1）。发现编码153个aa的Der p 39与人TnC和（Der f）39的同源性分别为35.63%和99.35%。IgE-ELISA显示，在5/87（5.75%）的HDM过敏血清样本中，IgE与表达并纯化的重组Der p 39（18 kDa）结合。对基于Der p 39的截短蛋白及杂交蛋白的IgE结合分析表明，IgE结合表位可能位于C端区域，且依赖于构象结构。本研究的数据已提交至世界卫生组织和国际免疫学会联盟（WHO/IUIS）过敏原命名数据库。
结论
Der p 39被鉴定为一种具有构象性IgE结合表位的次要HDM过敏原。这些发现可能对HDM过敏诊断和治疗的发展具有重要的理论意义。
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