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.

严重哮喘的了解和治疗仍知之甚少。近一半的严重哮喘患者 几乎没有证据表明传统 IgE 介导的过敏，但有强有力的证据表明混合适应性 皮质类固醇（CS）难治的免疫炎症过程。我们组公布的数据 显示蛋白质和 mRNA 水平上干扰素 γ (IFN)/Th1 通路激活增加 70% 的严重哮喘患者的支气管肺泡灌洗 (BAL) 细胞中，通常存在 与低水平的 2 型通路激活相关。这种复杂的免疫过程与 随着先天反应细胞因子 IL-27 表达的增加，增强硝基氧化作用 压力和低 IL-10 蛋白水平（项目 1）。在体外，将这些细胞因子组合在原代细胞上 人气道上皮细胞（HAEC）增强硝基氧化应激和炎症 通过增强 STAT1 的激活和减少 STAT3 的激活的途径， 导致硝基氧化应激和持续炎症。基于这样的背景， 我们假设活跃的 1 型和 2 型免疫途径与增强的 IL- 27、改变气道上皮中的 STAT1 和 3 激活平衡，导致增强 硝基氧化应激、炎症恶化和 CS 反应不良 疾病。为了解决这一假设，提出了以下 3 个具体目标： 目标 1。 在人类参与者中建立 1 型免疫与 2 型免疫过程相结合的情况 确定了与严重的 CS 反应性差的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. 确定以下因素的综合影响 1 型和-2 型细胞因子，在存在/不存在 IL-27 的情况下，对 HAEC 表型的体外影响。 确定 STAT1/STAT3 平衡和硝基氧化应激对这些影响的作用，并 目标 3. 在存在和存在的情况下确定 1/-2 型上皮的 CS 反应性 缺乏 IL-27 以及 IL-10 或抗氧化剂方法是否可以改善 CS 反应能力。该提案侧重于使用假设驱动的人类样本/疾病 以及与小鼠集成的无偏见（“组学”）方法和机械细胞培养工作 项目 1 中的模型。这些研究应该确定先天性和先天性交叉点的新途径。 和适应性免疫，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。