尘螨过敏原Der p 7新的IgE抗原表位鉴定及尘螨交叉反应性的研究  

Identification of new IgE antigenic epitopes of the dust mite allergen Der p 7,and investigation of dust mite cross⁃reactivity

作　　者：王璇 程露露[1] 胡婧 马克龙[1] 孙安源[3] 金腾川 陈凤 徐杨书函 WANG Xuan;CHENG Lulu;HU Jing;MA Kelong;SUN Anyuan;JIN Tengchuan;CHEN Feng;XU Yangshuhan(Anhui University of Traditional Chinese Medicine,Hefei 230012,China;Hefei National Laboratory for Physical Sciences at Microscale,CAS Key Laboratory of Innate Immunity and Chronic Disease,Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei 230027,China;The First Affiliated Hospital of the University of Science and Technology of China,Hefei 230036,China;Clinical Research Hospital of Chinese Academy of Sciences(Hefei),University of Science and Technology of China,Hefei 230001,China;College of Energy Materials and Chemical Engineering,Hefei University,Hefei 230000,China)

机构地区：[1]安徽中医药大学中西医结合学院,安徽合肥230012 [2]合肥微尺度物理科学国家实验室,中科院先天免疫与慢性病重点实验室,中国科学技术大学生命科学与医学部,安徽合肥230027 [3]中国科学技术大学第一附属医院,安徽合肥230036 [4]中国科学技术大学中国科学院(合肥)临床研究医院,安徽合肥230001 [5]合肥大学能源材料与化工学院,安徽合肥230000

出　　处：《海南医学院学报》2024年第22期1697-1707,共11页Journal of Hainan Medical University

基　　金：安徽中医药大学校级人才支持项目(DT2200000325);安徽省高校科研重点项目(RZ2300001713)。

摘　　要：目的:尘螨过敏原Der p 7新的IgE抗原表位鉴定及尘螨交叉反应性的研究。方法:ELISA检测华东地区安徽省合肥市的尘螨过敏患者与尘螨过敏临床相关性较高的重组蛋白Der p 7的IgE结合率;通过ELISA和Dot Blot鉴定屋尘螨过敏原Der p 7的新IgE抗原表位;利用圆二色光谱法检测Der p 7和其突变体的结构及热稳定性;最后用ELISA和ELISA抑制实验鉴定尘螨新的交叉过敏物质。结果:通过ELISA实验检测发现,华东地区的合肥市内尘螨过敏患者体内IgE与Der p 7的结合率为36.2%,与华南地区的广州市(37.4%)和西班牙(34.7%)相近。然而,这一结合率受到地理区域、自然环境和生活方式等多种因素的影响,与华北地区的北京市(19.3%)、乌克兰(22.67%)、意大利(28%)相比明显较低,而与非洲(56%)和日本(超过60%)相比则显著不同。ELISA和Dot Blot鉴定出尘螨重要过敏原Der p 7的两个新IgE抗原结合表位,分别是第36位天冬氨酸和第100位天冬氨酸。用圆二色光谱实验证明了Der p 7的结构稳定性,且突变后结构未发生改变,并检测了其热稳定性特征。最后用ELISA和ELISA抑制实验新发现了两种与尘螨存在交叉过敏的物质(小麦和花生)。结论:基于尘螨过敏原Der p 7与华东地区安徽省合肥市内过敏患者IgE的结合率及新的IgE抗原表位的研究为预防、诊断、治疗过敏性疾病提供理论基础,对开发尘螨过敏原低敏疫苗具有参考价值。发现新的尘螨交叉过敏物质为交叉过敏反应的预防、治疗提供了科学依据。Objective:To identify new IgE antigenic epitopes of the dust mite allergen Der p 7,and investigate dust mite cross‑reactivity.Methods:ELISA was used to detect the IgE binding of recombinant protein Der p 7 with high clinical relevance for dust mite allergy in dust mite‑allergic patients in Hefei City,Anhui Province,East China;new IgE antigenic epitopes of the house dust mite allergen Der p 7were identified by ELISA and Dot Blot;the structure and thermal stability of Der p 7 and its mutants were examined by circular dichroism spectrometry;finally,new cross‑allergens of dust mites were identified by ELISA and ELISA inhibition assay to identify new cross‑allergens of dust mites.Results:The binding rate of IgE to Der p 7 in dust mite allergic patients in Hefei City in East China was found to be 36.2%by ELISA,which was similar to that in Guangzhou City(37.4%)in South China and Spain(34.7%).However,this binding rate was influenced by a variety of factors such as geographic region,natural environment and lifestyle,and was significantly lower compared to Beijing(19.3%)in northern China,Ukraine(22.67%)and Italy(28%),and significantly different compared to Africa(56%)and Japan(more than 60%).Two new IgE binding epitopes of the dust mite major allergen Der p 7 were identified by ELISA and Dot Blot,which were aspartic acid at position 36 and aspartic acid at position 100,respectively.The structural stability of Der p 7 was demonstrated by circular dichroism spectroscopy experiments with no structural changes after mutation,and its thermal stability characteristics were examined.Finally we newly identified two substances(wheat and peanut)that are cross‑sensitised to dust mites using ELISA and ELISA inhibition experiments.Conclusion:Based on the binding rate of the dust mite allergen Der p 7 to the IgE of allergic patients in Hefei City,Anhui Province,East China,the study of new IgE antigenic epitopes provides a theoretical basis for the prevention,diagnosis,and treatment of allergic diseases,and is of reference valu

关 键 词：尘螨过敏 Der p 7 IgE结合率 IgE抗原表位 交叉过敏 

分 类 号：R446.6[医药卫生—诊断学] R593.1[医药卫生—临床医学]