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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：
10413107
负责人：
MARTIN D. CHAPMAN
金额：
$ 54.79万
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/10413107
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 Mammalian Cell Mediator of activation protein Mites Molecular Molecular Conformation Monoclonal Antibodies Multiple Myeloma Mus Mutate Nuclear Magnetic Resonance Patients Persons Phage Display Phase Play Prevalence Property Pulmonary Inflammation 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 rational design 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过敏原的抗原决定因素项目概要/摘要屋尘螨过敏是全球一个重要的健康问题，影响高达 85% 的哮喘儿童，以及因哮喘入院急诊的危险因素。第 1 类和第 2 类螨类过敏原占比较多螨过敏患者中屋尘螨特异性 IgE 反应性超过 50%。最近，Der p 23 还已被确定为主要的螨过敏原，但其对屋尘螨特异性 IgE 总量的贡献很小。第 1 组过敏原是半胱氨酸蛋白酶，会导致哮喘肺部炎症，而第 2 组过敏原过敏原是脂多糖结合蛋白。据报道，Der p 2 可以模仿人类结构- 通过收费样受体激活先天免疫系统的同系物。尽管有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 IgE 患病率高达 83% 在美国的螨过敏患者中都观察到了两组的 IgE 谱和与这两种主要过敏原相关的抗原决定簇尚不清楚。此次活动的主要目标项目是研究两组螨虫过敏原的抗原结构，以设计免疫疗法。过敏原特异性 IgE 单克隆抗体 (mAb) 将首次生产使用从融合获得的杂交瘤在体内发生重链和轻链的正确配对来自过敏性捐赠者和骨髓瘤伴侣的 B 细胞可赋予永生。过敏原将共结晶与重组 IgE 抗体构建体。参与 IgE 抗体结合的关键氨基酸是已识别并修改。具体目标是： 1) 分离针对哮喘相关过敏原的 IgE mAb 通过杂交瘤技术； 2) 通过 X 射线绘制第 1 组和第 2 组尘螨过敏原上的抗原决定簇 IgE 抗体结合表位的晶体学和分析； 3) IgE抗体的定点诱变表达具有 T 细胞反应性的低变应原性突变体的表位作为免疫治疗的候选者。一个人类库中螨虫过敏原特异性 IgE 抗体与螨虫过敏原之间关联的分析这些 IgE 抗体识别的表位将根据前两项中获得的信息进行目标。目标 #2 将生成 B 细胞表位三维结构的实验数据集，这些数据集是目前在用于开发 B 细胞表位预测工具的数据库中缺失。最重要的是，这项目将定义 IgE 抗体对螨虫过敏原的反应，并为合理的免疫反应提供结构基础。低过敏原的设计。在目标 #3 中，将通过以下方式分析与表位突变体结合的 IgE 抗体：将评估免疫测定和细胞介质释放测定以及 T 细胞反应性。突变体将是将选择比较和低变应原性形式用于螨免疫治疗疫苗的设计过敏。