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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变应原的抗原决定因素项目摘要/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，也是哮喘患者急诊室入院的危险因素。组1和组2变应原所占比例更高粉尘过敏患者总屋尘特异性IgE反应性的50%以上。最近，Der p 23也出现了已被确定为一种主要的粉尘变应原，但其对屋尘螨特异性IgE的贡献很小。第1组过敏原是导致哮喘肺部炎症的半胱氨酸蛋白酶，而第2组过敏原过敏原是脂多糖结合蛋白。据报道，DER P 2模仿了人类的结构- 通过Toll样受体激活先天免疫系统的同系物。尽管这些重要的分子蛋白水解组1与非蛋白水解组2的差异，过敏原高IgE患病率为83% 在美国的尘螨过敏患者中观察到了这两组的IgE。然而，IgE谱和与这两种主要过敏原相关的抗原决定簇尚不清楚。这样做的主要目标是本项目是为了研究两组尘螨变应原的抗原结构，为设计免疫疗法。过敏原特异性IgE单抗将首次与在体内发生的重链和轻链的正确配对，使用来自融合的杂交瘤来自过敏性捐献者的B细胞，与骨髓瘤伴侣一起获得永生。过敏原将共结晶用重组的IgE抗体构建。参与IgE抗体结合的关键氨基酸将是识别和修改。其具体目的是：1)分离哮喘相关变应原特异性IgE单抗通过杂交瘤技术；2)X射线定位第1、2组尘螨变应原的抗原决定簇 Ig E抗体结合表位的结晶学和分析；3)Ig E抗体的定点突变表达具有T细胞反应性的低变应原突变体的表位作为免疫治疗的候选者。一个尘螨变应原特异性免疫球蛋白E抗体与人类免疫缺陷的相关性分析这些IgE抗体识别的表位将使用在前两个中获得的信息来执行目标。目标2将生成B细胞表位的三维结构的实验数据集，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，这该项目将定义对尘螨变应原的IgE抗体反应，并将为Rational 低变应原的设计。在目标#3中，结合表位突变体的IgE抗体将通过以下方法进行分析免疫分析和细胞介体释放分析，以及T细胞反应性将被评估。变种人将会是将选择比较和低过敏性的形式来设计用于螨类免疫治疗的疫苗过敏。