Respiratory Enteroviruses, Inflammasome Activation and Innate Immune Cells

呼吸道肠道病毒、炎症小体激活和先天免疫细胞

• 批准号: 10299951
• 负责人: Marc B. Hershenson
• 金额: $ 26.1万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-10 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10299951
• 关键词: Acute; Acute respiratory infection; Adoptive Transfer; Adult; Aging; Allergens; Cells; Chemicals; Child; Clathrin; Data; Development; Disease Outbreaks; Endocytosis; Enterovirus; Enterovirus 68; Flow Cytometry; Fluorescence; Human; IL1R1 gene; Immune; Immune response; Immunoblotting; Immunofluorescence Immunologic; In Situ Hybridization; Infection; Inflammasome; Inflammation; Interleukin-1 beta; Interleukin-12; Interleukin-13; Interleukin-17; Interleukin-5; Knockout Mice; Lead; Link; Lung; Lymphoid Cell; Mediating; Modeling; Mus; Nose; Outcome; Play; Process; Production; Reporter; Respiratory Tract Infections; Rhinovirus; Rhinovirus infection; Role; TLR2 gene; TSLP gene; Testing; Time; Viral; Viral Genome; Viral Proteins; Virus; Virus Diseases; Work; airway hyperresponsiveness; airway inflammation; allergic airway disease; asthma exacerbation; cytokine; eosinophilic inflammation; inhibitor/antagonist; interleukin-23; macrophage; man; monocyte; neutrophil; receptor; respiratory; response; viral RNA

Project Summary Respiratory enteroviruses, including rhinoviruses (RVs) and enterovirus D68 (EV-D68), cause acute respiratory tract infections and asthma exacerbations. We previously identified the role of exudative macrophages in the development of airways inflammation and hyperresponsiveness (AHR) following RV-A1B infection. However, mechanisms by which macrophages contribute to airway responses have not been fully explored. We recently found that RV-A1B triggers Nod-like receptor protein 3 (NLRP3) inflammasome activation and IL-1β production in naïve and allergen-sensitized and challenged mice. Macrophages were required and sufficient for inflammasome activation, and NLRP3 null mice showed reduced airway inflammation and AHR, suggesting that macrophage NLRP3 inflammasome activation is required for RV-A1B-induced responses. EV-D68 caused an outbreak of severe respiratory illness in 2014 and is responsible for 14% of acute respiratory illnesses in children biannually. At the same time, RV-C was linked to severe respiratory infections and asthma exacerbations. Our pilot data show that, compared to RV-A1B, inflammasome activation and type 3 innate lymphoid cells (ILC3s) are increased in the lungs of EV-D68-infected mice. In contrast, inflammasome activation is decreased, and type 2 innate lymphoid cells (ILC2s) increased, in RV-C15-infected mice. We propose that the level of macrophage NLRP3 inflammasome activation and IL-1β produced by different respiratory enteroviruses determines their divergent innate immune cell responses and airway outcomes. EV-D68 triggers intense inflammasome activation and IL-1β-driven ILC3s, leading to IL-17-dependent neutrophilic inflammation. RV-C15 infection elicits minimal inflammasome activation, thus the predominant response is ILC2 expansion and IL-13-dependent eosinophilic inflammation. RV-A1B triggers an intermediate response. In Specific Aim 1, we will determine the mechanisms by which respiratory enteroviruses EV-D68, RV-A1B and RV-C15 differentially activate the macrophage NLRP3 inflammasome. We hypothesize that: 1) NLRP3 is required for EV-D68-induced inflammasome activation; 2) inflammasome priming is dependent on TLR2 and viral protein 4 (VP4); 3) RV-C inflammasome priming requires clathrin-mediated endocytosis; 4) EV-D68 and RV-A1B inflammasome activation is triggered by viral RNA; 5) RV-C15 viral genome is insufficient for inflammasome activation; and 6) EV-D68 viral protein 2B is sufficient for inflammasome activation. In Specific Aim 2, we will determine the role of the NLRP3 inflammasome in the activation of lung innate immune cells. We hypothesize that: 1) NLRP3 inflammasome activation is required for EV-D68-induced expansion of lung ILC3s, neutrophilic inflammation and AHR; 2) RV-C15-induced eosinophilic inflammation requires ILC2s; 3) in the context of allergic airways disease, IL-1β, in combination with IL-25 and IL-33, promotes enterovirus-induced ILC2 activation and eosinophilic inflammation; and 4) human infection with respiratory enteroviruses induces virus-specific inflammasome activation and expansion of nasal ILCs.

项目摘要 呼吸道肠道病毒，包括鼻病毒(RV)和肠道病毒D68(EV-D68)，可引起急性呼吸道 呼吸道感染和哮喘加重。我们先前确定了渗出性巨噬细胞在 RV-A1B感染后呼吸道炎症和高反应性(AHR)的发展然而， 巨噬细胞促进呼吸道反应的机制尚未完全探讨。 我们最近发现，RV-A1B可触发NLRP3的炎症小体激活和 幼稚和过敏原致敏和挑战的小鼠产生IL-1β。巨噬细胞是炎症小体激活所必需的，并且NLRP3基因缺失的小鼠表现出减少的呼吸道炎症和AHR，这表明巨噬细胞NLRP3炎症小体激活是RV-A1B诱导的反应所必需的。 EV-D68在2014年引起严重呼吸道疾病的暴发，每两年导致14%的儿童急性呼吸道疾病。与此同时，RV-C与严重的呼吸道感染和 哮喘加重。我们的试验数据显示，与RV-A1B相比，炎症体激活和3型 EV-D68感染小鼠肺内固有淋巴样细胞(ILC3)增多。相比之下，炎症性小体 在RV-C15感染的小鼠中，活性降低，2型固有淋巴样细胞(ILC2s)增加。 我们认为，不同的细胞因子对巨噬细胞炎性小体活化和IL-1β水平的影响 呼吸道肠道病毒决定了它们不同的先天免疫细胞反应和呼吸道结局。EV-D68触发强烈的炎症体激活和IL-1β驱动的ILC3，导致IL-17依赖的中性粒细胞 发炎。RV-C15感染引起轻微的炎性小体激活，因此主要反应是 ILC2扩张和IL-13依赖的嗜酸性炎症。RV-A1B可触发中间反应。 在具体目标1中，我们将确定呼吸道肠道病毒EV-D68， RV-A1B和RV-C15对巨噬细胞NLRP3炎症体的激活作用不同。我们假设： 1)NLRP3是EV-D68诱导的炎症体激活所必需的；2)炎症体的启动依赖于 TLR2和病毒蛋白4(VP4)；3)RV-C炎症体启动需要网状蛋白介导的内吞作用；4)EV-D68和RV-A1B炎症体激活是由病毒RNA触发的；5)RV-C15病毒基因组不足以 6)EV-D68病毒蛋白2B足以激活炎症小体。 在特定的目标2中，我们将确定NLRP3炎症体在肺激活中的作用 先天免疫细胞。我们假设：1)EV-D68诱导的肺ILC3的扩张、中性粒细胞炎症和AHR需要NLRP3炎症体的激活；2)RV-C15诱导的嗜酸性炎症需要ILC2s；3)在过敏性呼吸道疾病的背景下，IL-1β与IL-25和IL-33相结合， 促进肠道病毒诱导的ILC2激活和嗜酸性炎症；以及4)人类感染呼吸道肠道病毒诱导病毒特异性炎症小体激活和鼻部ILC扩张。