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主要犬过敏原 Can f 1 上构象 IgE 表位的聚类。

Clustering of conformational IgE epitopes on the major dog allergen Can f 1.

机构信息

Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.

Institute of Molecular Biosciences, University of Graz, Graz, Austria.

出版信息

Sci Rep. 2017 Sep 22;7(1):12135. doi: 10.1038/s41598-017-11672-5.

Abstract

Immunoglobulin E (IgE)-associated allergy affects more than 25% of the population. Can f 1 is the major dog allergen associated with respiratory symptoms but the epitopes recognized by allergic patients IgE on Can f 1 are unknown. To characterize IgE epitopes of Can f 1 recognized by dog allergic patients, six overlapping peptides spanning the Can f 1 sequence were synthesized. In direct IgE epitope mapping experiments peptides were analyzed for IgE reactivity by dot blot and Enzyme-linked immunosorbent assay (ELISA) with sera from dog allergic patients. For indirect epitope-mapping, rabbits were immunized with the peptides to generate specific IgG antibodies which were used to inhibit allergic patients' IgE binding to Can f 1. IgE binding sites were visualized on a model of the Can f 1 three-dimensional structure. We found that Can f 1 does not contain any relevant sequential IgE epitopes. However, IgE inhibition experiments with anti-peptide specific IgGs showed that Can f 1 N- and C-terminal portion assembled a major conformational binding site. In conclusion, our study is the first to identify the major IgE epitope-containing area of the dog allergen Can f 1. This finding is important for the development of allergen-specific treatment strategies.

摘要

免疫球蛋白 E(IgE)相关过敏影响超过 25%的人口。Can f 1 是与呼吸道症状相关的主要犬过敏原,但过敏患者 IgE 识别的 Can f 1 表位尚不清楚。为了鉴定过敏患者 IgE 识别的 Can f 1 的 IgE 表位,合成了六个跨越 Can f 1 序列的重叠肽。在直接 IgE 表位作图实验中,通过斑点印迹和酶联免疫吸附试验(ELISA)分析肽与犬过敏患者血清的 IgE 反应性。对于间接表位作图,用肽免疫兔子以产生特异性 IgG 抗体,并用其抑制过敏患者的 IgE 与 Can f 1 的结合。在 Can f 1 三维结构模型上观察 IgE 结合位点。我们发现 Can f 1 不含任何相关的连续 IgE 表位。然而,用抗肽特异性 IgG 进行的 IgE 抑制实验表明,Can f 1 的 N-和 C-末端组装了一个主要的构象结合位点。总之,我们的研究首次鉴定了犬过敏原 Can f 1 中包含主要 IgE 表位的区域。这一发现对开发过敏原特异性治疗策略很重要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dcf/5610169/244770fff9b6/41598_2017_11672_Fig1_HTML.jpg

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