Rhinovirus and Airway Epithelial Cell Responses

鼻病毒和气道上皮细胞反应

• 批准号: 7588762
• 负责人: Marc B. Hershenson
• 金额: $ 36.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-15 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7588762
• 关键词: 1-Phosphatidylinositol 3-Kinase; Accident and Emergency department; Accounting; Acute; Adult; Air; Allergic; Asthma; Biochemical; Bronchial Lavages; CXC Chemokines; Cell Culture Techniques; Cells; Child; Common Cold; Data; Disease; Enhancers; Epithelial; Epithelial Cells; Event; Extracellular Signal Regulated Kinases; Family Picornaviridae; Growth; Human; ICAM1 gene; Infection; Inflammation; Inflammatory; Inflammatory Response; Intercellular adhesion molecule 1; Interleukin-13; Interleukin-8; Lead; Life Cycle Stages; Ligation; Liquid substance; Mediating; Membrane Microdomains; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Mus; NADPH Oxidase; Nose; Oncogenes; Pathway interactions; Patients; Phosphorylation; Phosphotransferases; Pilot Projects; Production; RNA Viruses; Research Personnel; Rhinovirus; Signal Pathway; Signal Transduction; Site; Testing; Transactivation; Transcription Factor AP-1; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Viral; Virus Diseases; airway inflammation; base; chemokine; chemokine receptor; cytokine; in vivo; leucylarginine; macrophage inflammatory protein 2; neutrophil; p65; programs; promoter; response

Rhinovirus (RV) infection accounts for a large fraction of asthma exacerbations. Airway neutrophils and IL-8 levels are increased in RV-induced exacerbations, suggesting that RV stimulates exacerbations by inducing epithelial cell expression of (El_R)+ C-X-C chemokines, leading to an exaggerated inflammatory response. In pilot studies, we have shown that RV39 induces IL-8, ENA-78 and GRO-ct expression in primary, mu- cociliary-differentiated human tracheal epithelial cells. In 16HBE14o- cells, RV39 infection activates Src, PI 3-kinase, Akt and ERK minutes after infection, and activation of these kinases is required for IL-8 expression. RV increases C-X-C chemokine expression induced by two pro-asthmatic cytokines, IL-13 and TNFa. Fi- nally, RV1B infection of C57/BL6 mice increases airway neutrophils and levels of MIP-2, a murine ELR(+) C- X-C chemokine. Wetherefore hypothesize that RV is sufficient to activate biochemical signalingpathways involved in the asthmatic response, providing a mechanism for RV-induced asthma exacerbations. Specific Aim 1: Characterize upstream activators and downstream effectors of PI 3-kinase required for RV-induced ELR(+) C-X-C chemokine expression. We hypothesize that: 1) RV colocalizes with Src, PI 3- kinase, Akt and Grb2 in lipid rafts; 2) Src is required for activation of the PI 3-kinase/Akt pathway; 3) Class IA, II and III PI 3-kinases are required for maximal RV-induced expression of IL-8, ENA-78 and GROot; and 4) maximal NF-KB activation requires PI 3-kinase-dependent activation of NADPH oxidase. Specific Aim 2: Determine the biochemical signaling mechanisms responsible for cooperative effects of RV and pro-asthmatic cytokines on airway epithelial cell IL-8 expression. We hypothesize that: 1) ERKand JNK regulate IL-8 expression via activation of the AP-1 promoter site, which functions as a basal level en- hancer; 2) additive effects of RV39 and TNFa are mediated by increased p65 RelA phosphorylation and NF- transactivation; 3) synergistic effects of RV39 and IL-13 are mediated by increased AP-1 transactivation. Specific Aim 3: Determine the steps in the viral life cycle required or sufficient for RV-induced signaling and chemokine responses and, conversely, determine the requirement of host cell signal transduction for viralinfection. We hypothesize that: 1) ICAM1 ligation is required and sufficient for activation of Src, PI 3- kinase, Akt, ERK and JNK; 2) viral replication is not required for activation of these signaling intermediates; and 3) PI 3-kinase activation is required for RV39 internalization. Specific Aim 4: Determine the requirements of PI 3-kinase signaling and ELR(+) C-X-C chemokines for RV-inducedresponses in vivo. We hypothesize that: 1) RV1B infection is sufficient for airway inflammation and epithelial cell signaling in vivo; 2) PI 3-kinase is required for RV1B-induced airway inflammation in vivo; and 3) C-X-C chemokine receptor (CXCR)-2 regulates RV1B-induced airway inflammation in vivo. Understandina RV-induced asthma exacerbations will lead to improvements in the treatment of this disease.

鼻病毒（RV）感染占哮喘恶化的很大一部分。气道中性粒细胞和 IL-8 在 RV 诱导的病情加重中，其水平升高，表明 RV 通过诱导病情加重来刺激病情加重 (El_R)+ C-X-C 趋化因子的上皮细胞表达，导致炎症反应过度。 在初步研究中，我们已经表明 RV39 诱导原代、mu-中 IL-8、ENA-78 和 GRO-ct 表达 纤毛分化的人气管上皮细胞。在16HBE14o-细胞中，RV39感染激活Src、PI 感染后几分钟内出现 3-激酶、Akt 和 ERK，这些激酶的激活是 IL-8 表达所必需的。 RV 增加由两种促哮喘细胞因子 IL-13 和 TNFa 诱导的 C-X-C 趋化因子表达。菲- 最后，C57/BL6 小鼠的 RV1B 感染增加了气道中性粒细胞和 MIP-2（一种小鼠​​ ELR(+) C-）的水平 X-C趋化因子。因此我们假设 RV 足以激活生化信号通路 参与哮喘反应，为 RV 诱导的哮喘恶化提供机制。 具体目标 1：表征 PI 3-激酶所需的上游激活子和下游效应子 RV 诱导的 ELR(+) C-X-C 趋化因子表达。我们假设：1) RV 与 Src、PI 3- 共定位 脂筏中的激酶、Akt 和 Grb2； 2) Src是PI 3-激酶/Akt途径激活所必需的； 3) 班级 RV 诱导的 IL-8、ENA-78 和 GROot 最大表达需要 IA、II 和 III PI 3 激酶；和 4) 最大程度的 NF-KB 激活需要 NADPH 氧化酶的 PI 3 激酶依赖性激活。 具体目标 2：确定负责协同效应的生化信号机制 RV 和促哮喘细胞因子对气道上皮细胞 IL-8 表达的影响。我们假设：1）ERK 和 JNK 通过激活 AP-1 启动子位点来调节 IL-8 的表达，该位点作为基础水平的酶发挥作用。 汉瑟； 2) RV39 和 TNFa 的累加效应是通过增加 p65 RelA 磷酸化和 NF-介导的 反式激活； 3) RV39 和 IL-13 的协同作用是由 AP-1 反式激活增加介导的。 具体目标 3：确定病毒生命周期中 RV 诱导信号传导所需或充分的步骤 和趋化因子反应，并反过来确定宿主细胞信号转导的需求 病毒感染。我们假设：1) ICAM1 连接是必需的并且足以激活 Src、PI 3- 激酶、Akt、ERK 和 JNK； 2）这些信号中间体的激活不需要病毒复制； 3) RV39 内化需要 PI 3-激酶激活。 具体目标 4：确定 PI 3 激酶信号传导和 ELR(+) C-X-C 趋化因子的要求 RV 诱导的体内反应。我们假设：1) RV1B 感染足以引起气道炎症 和体内上皮细胞信号传导； 2) PI 3-激酶是RV1B诱导的体内气道炎症所必需的； 3) C-X-C趋化因子受体(CXCR)-2在体内调节RV1B诱导的气道炎症。 了解 RV 引起的哮喘加重将有助于改善这种疾病的治疗。