Epitope validation of fungal allergens in severe asthma

严重哮喘中真菌过敏原的表位验证

• 批准号: 10321625
• 负责人: Pandurangan Vijayanand
• 金额: $ 34.95万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321625
• 关键词: Admission activity; Adrenal Cortex Hormones; Adult; Allergens; Allergic; Allergic Bronchopulmonary Aspergillosis; Alternaria; Antigens; Aspergillus; Aspergillus fumigatus; Asthma; Automobile Driving; Bioinformatics; Biological Assay; Biological Response Modifier Therapy; Bronchiectasis; CD4 Positive T Lymphocytes; Cattle; Cells; Cessation of life; Child; Clinical; Cohort Studies; Coin; Common Epitope; Data Set; Dictyoptera; Enrollment; Epitopes; Exposure to; Failure; Food; Funding; Gene Expression Profile; Hospitalization; Inflammatory; Inhalation; Institutes; Label; Lead; Life; Milk; Molecular; Molecular Profiling; Mus; National Institute of Allergy and Infectious Disease; Nature; Oral; Outcome; Pathogenesis; Pathogenicity; Patients; Pertussis; Phenotype; Play; Production; Publishing; Pyroglyphidae; Recruitment Activity; Reproduction spores; Research; Risk; Role; Sampling; Severities; Severity of illness; Steroid Resistance; Steroids; Subgroup; T cell response; T-Lymphocyte; TNFSF5 gene; Testing; Tetanus; Th2 Cells; Time; United States National Institutes of Health; Vaccination; Validation; Visit; adverse outcome; airway inflammation; asthma exacerbation; asthmatic; asthmatic airway; base; chemokine; cohort; cytokine; data management; genome-wide; inner city; insight; neutrophil; pulmonary function; response; single-cell RNA sequencing; transcriptome sequencing; transcriptomics; validation studies

PROJECT SUMMARY In this application, we will characterize CD4 T cells reactive to epitopes of the fungal allergens Aspergillus fumigatus (ASP) and Alternaria alternata (ALT) to understand the molecular basis of severe asthma. Sensitization and/or exposure to antigens (spores) from fungal species such as ASP and ALT have been strongly associated with risk for a number of asthma-severity-related adverse outcomes. Based on this premise, we will test the hypotheses that (i) Fungal allergen-specific CD4 T cells are more pathogenic and qualitatively different when compared to allergen-specific CD4 T cells directed towards non-fungal inhaled allergens such as house dust mite (HDM), cockroach (CR), mouse (MO) or potential oral allergens such as cow milk (CM). (ii) Fungal allergen-epitope-specific CD4 T cells in patients with severe asthma have more pathogenic features than those from mild asthmatics. We will capitalize on subjects enrolled in three large asthma cohorts for these studies. In Aim 1, we will define epitopes recognized by Aspergillus (ASP) and Alternaria (ALT)-sensitized asthmatics categorized based on disease severity. The reactivity of predicted ASP and ALT epitopes will be tested in 20 sensitized subjects with mild and severe asthma and in 20 non-sensitized subjects without asthma to assess the nature of allergic versus healthy responses. By comparing the epitope-reactivity of the three cohorts, we will define epitopes that are common or specific to each subgroup: severe asthma, mild asthma and non- sensitized non-asthmatics. In Aim 2, we will determine the molecular profile of fungal (ASP and ALT)-specific CD4 T cells in patients with different asthma severity. We will utilize fungal epitope mega-pools to define the phenotype and transcriptional profile (at bulk and single-cell level) of ASP and ALT epitope-specific CD4 T cell responses in 10-20 fungal-sensitized subjects with severe and mild-to-moderate asthma, allergic bronchopulmonary aspergillosis (ABPA) and in control subjects without asthma. In parallel, we will also assess the molecular profile of fungal-specific CD4 T cells present in the airways of asthmatic subjects. To determine if the molecular features of fungal allergen-specific CD4 T cells are different from allergen-specific CD4 T cells directed to HDM, CR, MO, CM or antigen-specific Th2 cells induced by vaccination (pertussis and tetanus), we will perform additional transcriptomic analysis of these cells, and through the integrated Data Management Core specializing in bioinformatics cross compare data sets with Project 1 and 2. Together these fungal epitope validation studies in subjects with varying asthma severity will provide insights into the nature of CD4 T cell responses that drive pathogenesis of severe asthma.

项目摘要 在本申请中，我们将表征对真菌过敏原曲霉菌的表位反应的CD 4 T细胞。 烟曲霉菌（ASP）和互隔交链孢菌（ALT）以了解严重哮喘的分子基础。 致敏和/或暴露于来自真菌物种的抗原（孢子），如ASP和ALT， 与许多哮喘严重程度相关的不良后果的风险密切相关。基于此 在此前提下，我们将检验以下假设：（i）真菌变应原特异性CD 4 T细胞更具致病性， 与针对非真菌吸入的过敏原特异性CD 4 T细胞相比， 过敏原如屋尘螨（HDM）、蟑螂（CR）、小鼠（MO）或潜在的口腔过敏原如 牛奶（CM）(ii)严重哮喘患者的真菌变应原表位特异性CD 4 T细胞比正常人多 与轻度哮喘患者相比，我们将利用在三个大型研究中心注册的受试者， 这些研究的哮喘队列。 目的1：确定曲霉菌（ASP）和链格孢菌（ALT）致敏哮喘患者识别的表位 根据疾病严重程度分类。预测的ASP和ALT表位的反应性将在20个 轻度和重度哮喘的致敏受试者和20名无哮喘的非致敏受试者，以评估 过敏反应与健康反应的本质。通过比较三个队列的表位反应性，我们 将定义每个亚组的共同或特异性表位：重度哮喘，轻度哮喘和非哮喘。 敏感的非哮喘患者。在目标2中，我们将确定真菌（ASP和ALT）特异性的分子谱， 不同严重程度哮喘患者CD 4 T细胞的变化我们将利用真菌表位大库来定义 ASP和ALT表位特异性CD 4 T细胞的表型和转录谱（在整体和单细胞水平） 在10-20名真菌致敏的重度和轻中度哮喘受试者中的反应，过敏性 支气管肺曲霉菌病（ABPA）和对照组无哮喘。与此同时，我们还将评估 哮喘受试者气道中存在的真菌特异性CD 4 T细胞的分子谱。以确定是否 真菌变应原特异性CD 4 T细胞的分子特征不同于变应原特异性CD 4 T细胞 针对HDM、CR、MO、CM或疫苗接种（百日咳和破伤风）诱导的抗原特异性Th 2细胞，我们 将对这些细胞进行额外的转录组学分析，并通过集成的数据管理 核心专业生物信息学交叉比较数据集与项目1和2。这些真菌 在不同哮喘严重程度的受试者中进行的表位验证研究将提供对CD 4 T细胞性质的深入了解， 导致严重哮喘发病的细胞反应。