Molecular basis for anaphylaxis due to galactose-alpha-1,3-galactose (alpha-gal)

半乳糖-α-1,3-半乳糖（α-gal）引起的过敏反应的分子基础

基本信息

批准号：
10040328
负责人：
Scott Alan Smith
金额：
$ 24.03万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-02 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10040328
关键词：
Adaptive Immune System Affinity Allergens Allergic Allergic Disease Anaphylaxis Antibodies Antibody Response Antigens B-Lymphocytes Basophils Binding Biological Assay Blood Circulation Cells Clinical Clone Cells Column Chromatography Data Set Developed Countries Developing Countries Development Dideoxy Chain Termination DNA Sequencing Disease Epidemic Epitopes Exhibits Exposure to Food Gene Amplification Generations Genes Genetic Germ Lines Glycoproteins Goals Helminths Human Hybridomas Hypersensitivity IgE Immediate hypersensitivity Immunoglobulin Class Switching Immunoglobulin G Immunoglobulin Somatic Hypermutation Immunoglobulin Switch Recombination Immunohistochemistry Immunoprecipitation Incidence Individual Kinetics Knowledge Length Maps Mass Spectrum Analysis Measurement Meat Mediating Mediator of activation protein Methods Molecular Molecular Target Monoclonal Antibodies Mutate Oligosaccharides Pathogenesis Pathogenicity Pathologic Patients Peripheral Blood Mononuclear Cell Phylogeny Polysaccharides Population Process Reaction Recombinants Research Subjects Risk Serum Specificity Surface Plasmon Resonance Techniques Technology Testing Ticks Tissues Transgenic Mice Variant Western Blotting alpha-gal syndrome associated symptom base crosslink experience functional group galactosyl-(1-3)galactose human monoclonal antibodies immunoaffinity chromatography insight mast cell neoantigens novel peripheral blood receptor response vector tick

项目摘要

PROJECT SUMMARY The IgE molecule lies at the center of the pathogenesis of allergic diseases. In sensitized individuals, re-exposure to the offending allergen results in IgE engagement, causing Fcε receptor cross-linking and activation of mast cells and basophils. This triggers the release of mediators into the local tissue, resulting in the vast array of symptoms associated with allergic diseases, including anaphylactic shock. Studies of the human IgE molecule, and its targeted allergens, have been very limited. Nearly all of our knowledge of this process has come from studies using allergic patient serum, which contains a mixture of many antibodies, with many specificities, directed toward many different epitopes, and having many different affinities; thus studies of the molecular interactions of IgE with target allergens are greatly flawed. The ideal way to study this process is to use naturally- occurring human IgE monoclonal antibodies (mAbs), isolated from allergic subjects. Unfortunately, due to many impassible intrinsic technical hurdles no such antibodies have previously ever been made. We have established a method to grow, identify and immortalize IgE encoding B cells by making human hybridomas from the peripheral blood of allergic individuals. In this proposal, we develop the first panel of naturally-occurring alpha- gal-specific human IgE mAbs from subjects with red meat allergy. We intend to define the precise molecular basis for IgE-mediated reactions to red meat, from both the allergen and the antibody perspective. We will begin by generating human hybridomas from peripheral blood B cells of a few highly characterized research subjects. Purified IgE mAbs then will be used to precisely define critical molecular details of human anti-alpha-gal IgE binding. We will employ this panel of antibodies in various molecular assays to assist in localizing, identifying, and ultimately characterizing the allergen(s) found in ticks, responsible for sensitization of humans. As these IgE mAbs represent the B cells which are induced to undergo class-switch recombination, we will use their variable gene sequences to interrogate Illumina sequencing datasets from the research subjects from which the mAbs were obtained. This will allow for a keen understanding of the B cell population(s) underlying this allergic disease and may provide insights needed to predict individuals’ risk of sensitization. Together, this proposal will begin studies to define the exact glycoprotein targets of the human IgE B cell response, and the origin of the B cells themselves, which will provide much needed details underlying the sensitization and development of hypersensitivity to this common oligosaccharide.

项目摘要 IgE分子位于过敏性疾病发病机理的中心。在敏感的个体中，重新曝光对违规过敏原导致IgE参与，从而导致Fcε受体交联和桅杆的激活细胞和嗜碱性粒细胞。这触发了介体向当地组织的释放，导致了大量的与过敏性疾病有关的症状，包括过敏性休克。人类IgE分子的研究，它的目标过敏原非常有限。我们几乎所有对这一过程的了解都来自使用过敏患者精华素的研究，其中包含许多抗体的混合物，并具有许多特异性针对许多不同的表位，并具有许多不同的亲和力；因此研究分子 IgE与靶过敏原的相互作用存在很大缺陷。研究此过程的理想方法是自然使用 - 发生的人IgE单克隆抗体（mAb），从过敏性受试者中分离出来。不幸的是，由于很多以前从未有过这种抗体，固有的技术障碍没有。我们已经建立了一种生长，识别和永生化编码B细胞的方法，通过使人类杂交从过敏个体的外周血。在此提案中，我们开发了第一个天然α- 带有红肉过敏的受试者的gal特异性人Ige mab。我们打算定义精确的分子从过敏原和抗体的角度来看，IgE介导的对红肉的反应的基础。我们将开始通过从一些高度表征的研究对象的外周血B细胞中产生人类杂交瘤。然后将使用纯化的IgE mAb来精确定义人类抗阿尔帕 - 吉尔IgE的临界分子细节结合。我们将在各种分子测定中使用该小组抗体来帮助定位，识别，识别，并最终描述了在tick中发现的过敏原，负责人类的敏感性。如这些 mAb代表被诱导的B细胞进行类切换重组，我们将使用它们的变量从研究主题中询问Illumina测序数据集的基因序列获得了。这将使对这种过敏性疾病的B细胞种群有敏锐的了解并可能提供预测个人敏感性风险所需的见解。共同开始这个建议将开始定义人IgE B细胞反应的确切糖蛋白靶标的研究和B细胞的起源他们本身将提供急需的细节对这种常见的寡糖的高敏性。