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：哮喘相关过敏原的抗原决定因素用于设计免疫疗法项目摘要/摘要房屋尘螨过敏是全球重要的健康问题，影响多达85％的哮喘儿童，以及哮喘急诊室入院的危险因素。第1组和第2组螨过敏原占更多螨过敏患者中房屋尘螨特异性的总体粉尘特异性反应性的50％。最近，d d p 23也有被确定为主要的螨过敏原，但它对房屋尘螨特异性IgE的贡献很小。第1组过敏原是半胱氨酸蛋白酶，导致哮喘肺部炎症，而第2组过敏原是脂多糖结合蛋白。据报道，der p 2是模仿人类结构的同源物通过类似受体的收费来激活先天免疫系统。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％在美国的螨虫过敏患者中都观察到了这两组，但是，IgE曲目和与这两个主要过敏原相关的抗原决定因素尚不清楚。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）将首次生产使用从融合中获得的杂交瘤在体内纠正重链时，它们在体内发生来自过敏供体的B细胞与骨髓瘤伴侣赋予永生。过敏原将共结晶与重组IgE抗体构建体。 IgE抗体结合的关键氨基酸将是确定和修改。具体目的是：1）分离哮喘相关过敏原的IgE mAb特异性杂交瘤技术； 2）通过X射线在第1组和2组尘螨过敏原上映射抗原决定因素 IgE抗体结合表位的晶体学和分析； 3）IgE抗体的位置定向诱变表达具有T细胞反应性的低过敏性突变体的表达，作为免疫疗法的候选者。一个分析人类曲目的螨过敏原特异性IgE抗体与这些IgE抗体识别的表位将使用前两个获得的信息进行目标。 AIM＃2将生成B细胞表位三维结构的实验数据集，这是目前在用于开发用于B细胞表位预测工具的数据库中缺少。最重要的是，这项目将定义对螨过敏原的IgE抗体反应，并为理性提供结构性基础低过敏原的设计。在AIM＃3中，IgE抗体与表位突变体的结合将由免疫测定和细胞介质释放测定法，将评估T细胞反应性。突变体会将选择和低过敏性形式选择用于设计用于螨的免疫疗法的疫苗过敏。