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大豆过敏的免疫组学:表位的全面鉴定和特征分析。

The immunome of soy bean allergy: Comprehensive identification and characterization of epitopes.

机构信息

Ligand Development Unit, Fraunhofer IZI, Leipzig, Germany.

Molecular Biotechnology Division, Fraunhofer IME, Aachen, Germany.

出版信息

Clin Exp Allergy. 2019 Feb;49(2):239-251. doi: 10.1111/cea.13285. Epub 2018 Dec 3.

Abstract

BACKGROUND

The precise mapping of multiple antibody epitopes recognized by patients' sera allows a more detailed and differentiated understanding of immunological diseases. It may lead to the development of novel therapies and diagnostic tools.

OBJECTIVE

Mapping soy bean specific epitopes relevant for soy bean allergy patients and persons sensitized to soy bean, and analysis of their IgE/IgG binding spectrum.

METHODS

Identification of epitopes using sera, applying an optimized peptide phage display library followed by next-generation sequencing, specially designed in silico data analysis and subsequent peptide microarray analysis.

RESULTS

We were able to identify more than 400 potential epitope motifs in soy bean proteins. More than 60% of them have not yet been described as potential epitopes. Eighty-three peptides, representing the 42 most frequently found epitope candidates, were validated by microarray analysis using 50 sera from people who have been tested positive in skin prick test (SPT). Of these peptides, 56 were bound by antibodies, 55 by serum IgE, 43 by serum IgG and 30 by both. Person-specific epitope patterns were found for each individual and protein.

CONCLUSIONS

For individuals with clinical symptoms, epitope resolved analyses reveal a high prevalence of IgE binding to a few soy bean specific epitopes. Evaluation of individual immune profiles of patients with soy bean sensitization allows the identification of peptides that do facilitate studying individual IgE/IgG epitope binding patterns. This enables discrimination of sensitization from disease, such assay test has the potential to replace SPT assays.

摘要

背景

精确绘制患者血清中识别的多个抗体表位,可以更详细和差异化地了解免疫性疾病。这可能会导致新疗法和诊断工具的发展。

目的

绘制与大豆过敏患者和对大豆敏感的人相关的大豆特异性表位,并分析其 IgE/IgG 结合谱。

方法

使用血清,应用经过优化的肽噬菌体展示文库,然后进行下一代测序,专门设计的计算数据分析和随后的肽微阵列分析,来识别表位。

结果

我们能够在大豆蛋白中鉴定出 400 多个潜在的表位模体。其中超过 60%的尚未被描述为潜在的表位。通过使用 50 份经皮点刺试验(SPT)呈阳性的血清进行微阵列分析,验证了 83 个肽,代表 42 个最常发现的表位候选物。这些肽中有 56 个与抗体结合,55 个与血清 IgE 结合,43 个与血清 IgG 结合,30 个与两者都结合。为每个个体和蛋白质找到了个体特异性表位模式。

结论

对于有临床症状的个体,表位解析分析显示对少数大豆特异性表位的 IgE 结合具有高患病率。对大豆致敏患者的个体免疫谱进行评估,可以识别出有助于研究个体 IgE/IgG 表位结合模式的肽。这使得能够区分致敏和疾病,这种检测方法有可能取代 SPT 检测。

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