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

严重哮喘仍然知之甚少，治疗。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，但有强有力的证据表明混合适应性过敏， 皮质类固醇（CS）难治性免疫炎症过程。我们小组公布的数据 显示在蛋白质和mRNA水平上增加的干扰素γ（IFN γ）/Th 1途径活化 在70%的严重哮喘患者的支气管肺泡灌洗（BAL）细胞中， 与低水平的2型途径激活有关。这个复杂的免疫过程 随着先天性应答细胞因子IL-27表达的增加， 应激和低IL-10蛋白水平（项目1）。在体外，联合这些细胞因子对原发性 人气道上皮细胞（HAECs）增加硝基氧化应激和炎症反应 通过增强STAT 1的激活和减少STAT 3的激活， 导致硝基氧化应激和持续性炎症。基于这一背景， 我们假设，与增强的IL-1相关的活跃的1型和2型免疫途径， 27，改变气道上皮中STAT 1和STAT 3的激活平衡，导致气道上皮细胞中STAT 1和STAT 3的表达增加。 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3个具体目标如下：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 鉴定了与严重CS反应性差的哮喘相关的分子表型， IL-27的存在进一步改变了表型。目标2.确定以下因素的综合影响 在存在/不存在IL-27的情况下，1型和2型细胞因子对体外HAEC表型的影响。 确定STAT 1/STAT 3平衡和硝基氧化应激在这些效应中的作用， 目标3：在存在和不存在CS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 响应能力。该提案重点关注使用假设驱动的人类样本/疾病 和无偏见的（“组学”）方法和机械细胞培养工作与小鼠 项目1中的模型。这些研究应该确定新的途径，在先天的交叉点， 和适应性免疫，以更好地定义和表征严重哮喘及其表型， 并为治疗提供新的靶点。

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.