Project 2 Impact of Innate and Adaptive Immunity At the Airway Epithelium in Severe Asthma

项目 2 先天性和适应性免疫对严重哮喘气道上皮的影响

• 批准号: 8853017
• 负责人: Sally E Wenzel
• 金额: $ 38.21万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-06-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8853017
• 关键词: Address; Adrenal Cortex Hormones; Antigen-Antibody Complex; Antioxidants; Asthma; Autoimmune Process; Bronchoalveolar Lavage; Cell Culture Techniques; Cells; Clinical; DNA Binding; Data; Development; Disease; Dose; Epithelial; Epithelial Cells; Epithelium; Equilibrium; Gene Expression; Granulomatous; Human; Hypersensitivity; IgE; Immune; Immunity; In Vitro; Inflammation; Inflammatory; Interferon Type II; Interferons; Interleukin-10; Interleukin-13; Investigation; Measures; Mediating; Messenger RNA; Natural Immunity; Outcome; Oxidative Stress; Participant; Pathway interactions; Patients; Pattern; Phenotype; Process; Proteins; Publishing; Refractory; Reporting; Role; STAT1 gene; STAT3 gene; Sampling; Steroids; Stress; Work; adaptive immunity; airway epithelium; asthmatic; asthmatic patient; base; clinical phenotype; cytokine; improved; molecular phenotype; mouse model; novel; response; targeted treatment; transcriptome sequencing

Severe asthma remains poorly understood and treated. Nearly half of severe asthma patients have little evidence for traditional IgE mediated allergy but strong evidence for a mixed adaptive immune-inflammatory process refractory to corticosteroids (CS). Published data from our group show increased interferon gamma (IFN)/Th1 pathway activation at the protein and mRNA level in bronchoalveolar lavage (BAL) cells in 70% of patients with severe asthma, commonly associated with low level Type-2 pathway activation. This complex immune process associates with increased expression of the innate-responding cytokine IL-27 enhanced nitro-oxidative stress and low IL-10 protein levels (Project 1). In vitro, combination these cytokines on primary human airway epithelial cells (HAECs) augments nitro-oxidative stress and inflammatory pathways through enhanced activation of STAT1 and diminished activation of STAT3, contributing to nitro-oxidative stress and persistent inflammation. Based on this background, we hypothesize that active Type- 1 and -2 immune pathways, in association with enhanced IL- 27, alter the STAT1 and 3 activation balance in the airway epithelium leading to augmented nitro-oxidatvie stress, worsened inflammation and the presence of a poorly CS responsive disease. To address this hypothesis, 3 specific aims are proposed as follows: Aim 1. Establish in human participants that Type-1 immunity combined with Type-2 immune processes identifies a molecular phenotype associated with severe poorly CS responsive asthma and that the presence of IL-27 further worsens the phenotype. Aim 2. Identify the combined effect of Type-1 and -2 cytokines, in the presence/absence of IL-27, on HAEC phenotype in vitro. Determine the role of STAT1/STAT3 balance and nitro-oxidative stress on these effects and Aim 3. Determine the CS responsiveness of the Type-1/-2 epithelium, in the presence and absence of IL-27 and whether IL-10 or anti-oxidant approaches can improve the CS responsiveness. This proposal focuses on human samples/disease using hypothesis driven and unbiased (‘omics’) approaches and mechanistic cell culture work integrated with mouse models in Project 1. These studies should identify novel pathways at the intersection of innate and adaptive immunity to better define and characterize severe asthma and its phenotypes, as well as to provide new targets for the therapy.

对严重哮喘的认识和治疗仍然很少。近一半的重症哮喘患者 几乎没有证据表明传统的IgE介导的过敏反应，但有强有力的证据表明混合适应性过敏 对皮质类固醇(CS)无效的免疫炎症过程。来自我们小组的已发布数据 在蛋白质和基因水平显示干扰素/Th1途径的激活增加 在70%的重症哮喘患者的支气管肺泡灌洗(BAL)细胞中，通常 与低水平的2型通路激活有关。这一复杂的免疫过程与 与生俱来的细胞因子IL-27的表达增加促进了硝基氧化 压力和低IL-10蛋白水平(项目1)。在体外，将这些细胞因子结合在一起 人呼吸道上皮细胞(HAECs)增加硝基氧化应激和炎症反应 通过STAT1的增强激活和STAT3的减弱激活的通路， 导致硝基氧化应激和持续性炎症。基于这样的背景， 我们假设，活跃的1型和2型免疫通路，与增强的IL-1- 27，改变气道上皮中STAT1和STAT3的激活平衡，导致增强 硝基-氧化应激，炎症加重，CS反应性差 疾病。针对这一假设，本文提出了三个具体目标：目标1。 在人类参与者中建立1型免疫与2型免疫过程相结合 确定与严重的CS反应不良哮喘相关的分子表型 IL-27的存在进一步恶化了表型。目标2.确定以下各项的综合效果 IL-27存在或不存在时，1型和2型细胞因子对体外HAEC表型的影响。 确定STAT1/STAT3平衡和硝基氧化应激在这些效应中的作用 目的3.测定1/2型上皮细胞对CS的反应性 IL-27的缺失及IL-10或抗氧化剂能否改善CS 响应性。该建议使用假设驱动，重点关注人类样本/疾病 和不偏不倚的(组学)方法和机械细胞培养工作与小鼠相结合 项目1中的模型。这些研究应该确定先天的交叉点上的新途径 和获得性免疫，以更好地定义和表征严重哮喘及其表型，如 并为治疗提供新的靶点。