Impact of Airway Epithelium on Innate and Adaptive Immunity in the Lung

项目总结研究项目2 呼吸道病毒，如呼吸道合胞病毒(RSV)，是#年哮喘恶化的主要诱因。 无论是孩子还是成年人。该项目的主要假设是，呼吸道上皮细胞(AECs)协调 对呼吸道病毒感染和空气变应原的反应。此外，血管内皮细胞之间的内在差异 哮喘和健康儿童在感染和暴露于 过敏原。我们推测呼吸道合胞病毒或心率变异性哮喘患儿的血管内皮细胞感染 与过敏原暴露相结合，诱导常驻固有细胞，包括树突状细胞(DC)和 先天淋巴样细胞(ILCs)，以触发2型反应。我们将使用从以下来源分离的AEC来测试此想法 哮喘和健康儿童，生长在空气/液体界面(ALI)，并在 过敏原挑战的背景。我们将研究DC和ILCs与这些AEC共培养后的反应，以及 它们通过以下目的在促进2型炎症中发挥作用： 目的1.验证AECs控制局部DC成熟和功能的假说。由于AECS在 哮喘患者可能会因屏障缺陷而“漏气”，我们将首先确定是否有更多的抗原通过 使用改良的RSV株，哮喘患者的AECs比健康的AECs对DC的作用更强。然后，我们将检测哮喘患者的血管内皮细胞 儿童在促进DC分化和功能以及病毒感染和过敏原暴露如何影响方面存在差异 这种互动。最后，我们将评估哮喘儿童接触AECs的DC驱动CD4的能力 共培养实验中T细胞的分化和增殖。 目的2.测试假设在过敏原暴露和病毒感染后，哮喘和健康 AECS通过脂质介质和血管活性肠肽对肺ILC2产生不同程度的影响。我们会 评估血管内皮细胞在调节ILC2中的作用，ILC2是肺中含量最丰富的ILC。我们希望能找到 哮喘患儿的血管内皮细胞在病毒感染和过敏原攻击时增强ILC2功能。在我们的 初步数据，我们发现了两个不同的人肺ILC2亚群，它们表达不同的受体 氧甾醇、维甲酸和血管活性肠肽。因此，我们建议定义这些脂类药物的影响 以及神经肽介质对不同ILC2群体的激活作用。 目的3.验证ICOS ILCs调节肺免疫反应的假说。我们写了这部小说 观察到肺中含有不同寻常的ICOS ILC亚群。这些细胞不适合ILC1、ILC2、 ILC3范例，表达调节性淋巴细胞的标志，并富含免疫抑制 关联的环境。因此，我们假设这些细胞可能具有调节功能。为了测试这一点，我们 建议分离肺ICOS ILCs并确定其体外转录组及其功能 体外能力，包括细胞因子和趋化因子的分泌。此外，我们建议测试他们的 对暴露于HDM和/或病毒的ALI细胞刺激的反应性。

PROJECT SUMMARY RESEARCH PROJECT 2 Respiratory viruses, such as respiratory syncytial virus (RSV), are major triggers of asthma exacerbation in children and adults. The overarching hypothesis of this project is that airway epithelial cells (AECs) coordinate responses to respiratory virus infection and aeroallergens. Moreover, intrinsic differences between AECs of asthmatic and healthy children change both innate and adaptive responses during infection and exposure to allergens. We hypothesize that infection of AECs from asthmatic children with RSV or HRV, in combination with allergen exposure, induces resident innate cells, including dendritic cells (DCs) and innate lymphoid cells (ILCs), to trigger type-2 responses. We will test this idea using AECs isolated from asthmatic and healthy children, grown at air/liquid interface (ALI) and infected with respiratory viruses in the context of allergen challenge. We will examine the response of DCs and ILCs co-cultured with these AECs, and their role in promoting type-2 inflammation through the following aims: Aim 1. Test hypothesis that AECs control local DC maturation and function. Since AECs in asthmatics may be “leaky” due to barrier defects, we will first determine whether more antigen is transferred by asthmatic than healthy AECs to DCs using modified RSV strains. Then, we will test whether AECs from asthmatic children differ in promoting DC differentiation and function, and how virus infection and allergen exposure impact this interaction. Finally, we will assess the ability of DCs exposed to AECs from asthmatic children to drive CD4 T cell differentiation and proliferation in co-culture assays. Aim 2. Test hypothesis that after allergen exposure and viral infections, asthmatic and healthy AECs differentially impact lung ILC2 through lipid mediators and vasoactive intestinal peptide. We will assess the role of AECs in regulating ILC2s, which are the most abundant ILCs in the lung. We expect to find that AECs from asthmatic children enhance ILC2 function during viral infection and allergen challenge. In our preliminary data, we found two distinct subsets of human lung ILC2s that differentially express receptors for oxysterols, retinoic acid, and vasoactive intestinal peptide. Thus, we propose to define the impact of these lipidic and neuropeptide mediators on the activation of different ILC2 populations. Aim 3. Test hypothesis that ICOS+ ILCs regulate lung immune responses. We made the novel observation that the lung contains an unusual subset of ICOS+ ILCs. These cells do not fit into the ILC1, ILC2, ILC3 paradigm, express a marker of regulatory lymphocytes and are enriched in an immunosuppressive environment associated. Thus, we hypothesize that these cells may have regulatory functions. To test this, we propose to isolate lung ICOS+ ILCs and define their transcriptome profile ex vivo as well as their functional capabilities in vitro, including cytokine and chemokine secretion. Moreover, we propose to test their responsiveness to stimuli derived from ALI cultures exposed to HDM and/or viruses.