Role of the Respiratory Syncytial Virus Fusion Protein in Airway Mucus Induction

呼吸道合胞病毒融合蛋白在气道粘液诱导中的作用

基本信息

批准号：
8462893
负责人：
Martin Lawrence Moore
金额：
$ 35.98万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-15 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8462893
关键词：
Amino Acids Automobile Driving Basophils Biological Assay Bronchiolitis Cell Line Cells Cessation of life Chimeric Proteins Data Epithelial Epithelial Cells Functional disorder Genes Giant Cells Goals Hospitalization Human Hypoxia Immune response In Vitro Inbred BALB C Mice Infant Infection Inflammatory Interleukin-13 Interleukin-4 Laboratories Lead Lower respiratory tract structure Lung Measures Mechanical ventilation Mediating Modeling Molecular Models Mucins Mucous body substance Mus Mutagenesis Neutrophilia Pathogenesis Play Production Proteins Respiratory Syncytial Virus Infections Respiratory syncytial virus Role System T-Lymphocyte Transfection Viral Viral Bronchiolitis Viral Fusion Proteins Viral Load result Virulence Virulent Virus Virus Diseases airway hyperresponsiveness airway obstruction base cell injury cell type cytokine in vivo innovation molecular modeling mutant neutrophil positional cloning public health relevance research study response virus pathogenesis

项目摘要

DESCRIPTION (provided by applicant): Our long term goal is to elucidate mechanisms by which respiratory syncytial virus (RSV) causes lung dysfunction. In the USA, RSV hospitalizes >100K infants/year. Pulmonary mucus is a hallmark of RSV disease. Mucus mixed with epithelial cell debris blocks the airways. Mechanisms by which RSV infection induces airway mucus are not known. Goals of this proposal are to define the role of the viral fusion (F) proteins of mucus-inducing RSV strains in pathogenesis and to define the roles of neutrophils and basophils in RSV-induced mucus. In contrast to laboratory RSV strains, RSV strains line 19 and 2-20 induce mucus and bronchiolitis in mice. Using a RSV reverse genetics system, we showed that the F protein of line 19 plays a role in mucus induction. We identified five amino acids in the line 19 F protein as candidates for involvement in mucus induction. We will use F mutant viruses to define line 19 F residues that are important for mucus expression in vivo. RSV 2- 20 is virulent in mice. We hypothesize that RSV 2-20 F contributes to airway mucus and virulence. We will define the role of 2-20 F in pathogenesis using 2-20F-chimeric RSV. Line 19 and 2-20 cause greater epithelial cell cytopathic effect (CPE) and desquamation in vivo than lab RSV strains. We will use F mutants to define effects of RSV line 19 F and 2-20 F on epithelial cell CPE, syncytia, and desquamation in vivo and in vitro. We will define the fusogenicity of RSV line 19, 2-20, and lab strain F proteins in epithelial cells in vitro using a fusion assay. We hypothesize that line 19 F and 2-20 F induce greater syncytia and fusion in vivo and in vitro. The study will also determine the roles of neutrophils and basophils in RSV bronchiolitis. Concurrently wih epithelial cell damage, RSV line 19 and 2-20 induce neutrophilia in the lungs of mice. We hypothesize that neutrophils contribute to viral clearance and mucus expression in line 19 and 2-20 pathogenesis. Line 19- induced mucus is dependent on IL-13, a TH2 cytokine. IL 4 is the primary TH2-driving cytokine. We showed that basophils are the major IL 4-expressing cell type in RSV-infection. We hypothesize that basophils contribute to RSV line 19- and 2-20-induced IL-13 and mucus expression. These RSV bronchiolitis models will advance our understanding of RSV strain-dependent pathogenesis and may lead to much-needed therapies.

描述（由申请人提供）：我们的长期目标是阐明呼吸综合病毒（RSV）引起肺功能障碍的机制。在美国，RSV每年住院> 100k婴儿。肺粘液是RSV疾病的标志。与上皮细胞碎片混合的粘液阻断了气道。 RSV感染引起气道粘液的机制尚不清楚。该建议的目标是定义粘液诱导RSV菌株在发病机理中的病毒融合（F）蛋白的作用，并定义中性粒细胞和嗜碱性粒细胞在RSV诱导的粘液中的作用。与实验室RSV菌株相反，RSV菌株19和2-20诱导小鼠粘液和细支气管炎。使用RSV反向遗传学系统，我们表明第19行的F蛋白在粘液诱导中起作用。我们在第19 f蛋白中鉴定出五个氨基酸是参与粘液诱导的候选物。我们将使用F突变病毒来定义对体内粘液表达很重要的线19 f残基。 RSV 2-20在小鼠中有力。我们假设RSV 2-20 F有助于气道粘液和毒力。我们将使用2-20F-芝麻RSV来定义2-20 F在发病机理中的作用。第19行和第2-20行引起的上皮细胞细胞病变效应（CPE）和体内脱离比实验室RSV菌株。我们将使用F突变体来定义RSV线19 F和2-20 F对上皮细胞CPE，合成症和体外和体外的脱离的影响。我们将使用融合测定法定定义RSV系19、2-20和LAB菌株F蛋白的融合性菌株F蛋白。我们假设这条线19 F和2-20 F会引起更大的合成和体内融合。该研究还将确定嗜中性粒细胞和嗜碱性粒细胞在RSV细支气管炎中的作用。同时使用上皮细胞损伤，RSV第19和2-20系在小鼠的肺中诱导嗜中性粒细胞。我们假设中性粒细胞在第19行和2-20发病机理中有助于病毒清除和粘液表达。线19诱导的粘液取决于IL-13，Th2细胞因子。 IL 4是主要的Th2驱动细胞因子。我们表明，嗜碱性嗜碱性是RSV感染中主要的IL 4表达细胞类型。我们假设嗜碱性粒细胞有助于RSV线19和2-20诱导的IL-13和粘液表达。这些RSV支气管炎模型将提高我们对RSV菌株依赖性发病机理的理解，并可能导致急需的疗法。