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.

严重的哮喘知识和治疗程度仍然很差。近一半的严重哮喘患者 几乎没有证据表明传统的IGE介导的过敏，但有力的证据表明混合适应性 免疫炎性过程对皮质类固醇（CS）的难治性。来自我们小组的数据 显示在蛋白质和mRNA水平上激活干扰素γ（IFN）/Th1途径的增加 在70％的严重哮喘患者中，在支气管肺泡灌洗（BAL）中，通常 与低级2型途径激活相关。这个复杂的免疫过程伴侣 随着先天反应的细胞因子IL-27的表达增加增强了硝基氧化 压力和低IL-10蛋白水平（项目1）。体外，这些细胞因子在原发性上的结合 人气道上皮细胞（HAEC）增加了硝基氧化应激和炎症 通过增强STAT1激活的途径，STAT3的激活减少， 有助于硝基氧化应激和持续注射。基于此背景 我们假设主动1和-2免疫病与IL-相关的IL- 27，改变气道上皮的STAT1和3激活平衡，导致增强 硝基氧化应激，炎症恶化和CS响应不良的存在 疾病。为了解决这一假设，提出了3个具体目标，如下所示：目标1。 在人类参与者中建立1型免疫原结合2型免疫过程 确定与严重CS不良反应性哮喘相关的分子表型，并且 IL-27的存在进一步恶化了表型。目标2。确定的综合效果 在体外HAEC表型上，在存在/不存在IL-27的情况下，1型和-2个细胞因子。 确定STAT1/STAT3平衡和硝基氧化应力对这些影响的作用， 目标3。在存在下确定1/-2上皮的CS响应性 没有IL-27以及IL-10还是抗氧化剂方法可以改善CS 响应能力。该提案的重点是使用假设驱动的人类样本/疾病 和无偏见的方法和机械细胞培养物与小鼠整合 项目1中的模型。这些研究应确定先天交集的新途径 以及适应性免疫学以更好地定义和表征严重的哮喘及其表型，例如 并为治疗提供新的靶标。