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Antigenic determinants of asthma-associated allergens for design of immunotherapy

用于免疫治疗设计的哮喘相关过敏原的抗原决定簇

基本信息

批准号：
10196971
负责人：
MARTIN D. CHAPMAN
金额：
$ 58.48万
依托单位：
INDOOR BIOTECHNOLOGIES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-25 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10196971
关键词：
Accident and Emergency department Admission activity Affect Allergens Allergic Allergic Disease Amino Acids Antibodies Antibody Repertoire Antibody Response Asthma Award B-Lymphocyte Epitopes B-Lymphocytes Binding Biological Assay Canis familiaris Caspase Cells Characteristics Child Clinical Research Complex Crystallization Data Data Set Databases Dermatophagoides farinae antigen f 1 Dermatophagoides pteronyssinus antigen p 1 Dermatophagoides pteronyssinus antigen p 2 Development Dictyoptera Disease Engineering Epitope Mapping Epitopes Felis catus Funding Future Goals Grant Health Homologous Gene House Dust Mite Allergens Human Hybridomas Hypersensitivity IgE Immune Immune response Immunoassay Immunoglobulin G Immunologics Immunotherapeutic agent Immunotherapy Innate Immune System Libraries Light Lung Inflammation Mammalian Cell Mediator of activation protein Mites Molecular Molecular Conformation Monoclonal Antibodies Multiple Myeloma Mus Mutate Nuclear Magnetic Resonance Patients Phage Display Phase Play Prevalence Property Pyroglyphidae Recombinant Antibody Recombinant Proteins Recombinants Reporting Risk Factors Role Site-Directed Mutagenesis Source Specificity Structure T-Cell Activation T-Cell Proliferation T-Lymphocyte Technology Testing Time Toll-like receptors Vaccine Design Vaccines Variant X-Ray Crystallography asthmatic asthmatic patient base cockroach allergen combinatorial cytokine design dust mite allergy immunoreactivity immunotherapy trials impaired capacity improved in vivo indoor allergen inner city innovation lipopolysaccharide-binding protein mutant response three dimensional structure tool vaccine immunotherapy

项目摘要

TITLE: Antigenic determinants of asthma-associated allergens for design of immunotherapy PROJECT SUMMARY/ABSTRACT House dust mite allergy is an important health problem worldwide, affecting up to 85% of asthmatic children, and a risk factor for emergency room admission with asthma. Group 1 and 2 mite allergens account for more than 50% of total house dust mite specific IgE reactivity in mite allergic patients. Recently, Der p 23 has also been identified as a major mite allergen, but its contribution to the total house dust mite specific IgE is small. Group 1 allergens are cysteine proteases that contribute to lung inflammation in asthma, whereas group 2 allergens are lipopolysaccharide binding proteins. Der p 2 has been reported to mimic a human structural- homolog that activates the innate immune system through toll like receptors. Despite these important molecular differences between proteolytic group 1 and non-proteolytic group 2, a high IgE prevalence of 83% to allergens from both groups has been observed in mite allergic patients in the U.S. However, the IgE repertoire and antigenic determinants associated with these two major allergens are not known. The main goal of this project is to investigate the antigenic structure of both groups of mite allergens for the design of immunotherapy. Allergen-specific IgE monoclonal antibodies (mAb) will be produced for the first time with the correct pairing of the heavy and light chains as they occur in vivo, using hybridomas obtained from the fusion of B cells from allergic donors with a myeloma partner that confers immortality. Allergens will be co-crystallized with recombinant IgE antibody constructs. The key amino acids involved in IgE antibody binding will be identified and modified. The specific aims are: 1) Isolation of IgE mAb specific for asthma-associated allergens by hybridoma technology; 2) mapping of antigenic determinants on groups 1 and 2 dust mite allergens by X-ray crystallography and analysis of IgE antibody binding epitopes; and 3) site-directed mutagenesis of IgE antibody epitopes for expression of hypoallergenic mutants with T cell reactivity as candidates for immunotherapy. An analysis of the association between mite allergen-specific IgE antibodies from the human repertoire and the epitopes recognized by these IgE antibodies will be performed with the information obtained in the first two aims. Aim #2 will generate experimental data sets of three-dimensional structures of B-cell epitopes, which are currently missing in databases used for developing tools for B cell epitope prediction. Most importantly, this project will define IgE antibody responses to mite allergens and will provide the structural basis for rational design of hypoallergens. In Aim #3, IgE antibody binding to the epitope mutants will be analyzed by immunoassays and cell mediator release assays, and T cell reactivity will be evaluated. Mutants will be compared and hypoallergenic forms will be selected for the design of vaccines for immunotherapy of mite allergy.

标题：用于免疫治疗设计的哮喘相关变应原的抗原决定簇项目总结/摘要屋尘螨过敏是世界范围内的一个重要健康问题，影响高达85%的哮喘儿童，也是哮喘患者进急诊室的危险因素组1和2螨过敏原占更多尘螨过敏患者中总屋尘螨特异性IgE反应性的50%以上。最近，Der p 23还已被确定为一个主要的螨过敏原，但其贡献的总屋尘螨特异性IgE是小的。第1组过敏原是半胱氨酸蛋白酶，可导致哮喘患者肺部炎症，而第2组过敏原是半胱氨酸蛋白酶，过敏原是脂多糖结合蛋白。据报道，Derp 2模拟了人类的结构- 通过Toll样受体激活先天免疫系统的同源物。尽管这些重要的分子蛋白水解组1和非蛋白水解组2之间的差异，过敏原的高IgE患病率为83% 在美国的螨过敏患者中观察到了来自两组的IgE。与这两种主要过敏原相关的抗原决定簇尚不清楚。这个的主要目标项目是研究两组螨过敏原的抗原结构，免疫疗法过敏原特异性IgE单克隆抗体（mAb）将首次使用当重链和轻链在体内发生时，使用从以下融合获得的杂交瘤，来自过敏性捐赠者的B细胞与骨髓瘤伴侣，赋予永生。过敏原会被共结晶重组IgE抗体构建体。参与IgE抗体结合的关键氨基酸将是识别和修改。具体目的是：1）分离特异性针对哮喘相关变应原的IgE mAb 通过杂交瘤技术; 2）通过X射线对第1组和第2组尘螨变应原的抗原决定簇作图 IgE抗体结合表位的晶体学和分析;和3）IgE抗体的定点诱变表位用于表达具有T细胞反应性的低变应原性突变体作为免疫治疗的候选物。一个分析来自人类库的螨变应原特异性IgE抗体与这些IgE抗体识别的表位将用前两个步骤中获得的信息进行目标。目标#2将生成B细胞表位的三维结构的实验数据集，其是目前在用于开发B细胞表位预测工具的数据库中缺失。最重要的是这该项目将定义对螨过敏原的IgE抗体反应，并将为合理的低过敏原的设计。在目标#3中，将通过以下方法分析IgE抗体与表位突变体的结合：将评估免疫测定和细胞介质释放测定以及T细胞反应性。变种人将会比较和低变应原性形式将被选择用于设计用于螨的免疫治疗的疫苗过敏