Rhinovirus Pathogenesis and Host Range

鼻病毒发病机制和宿主范围

• 批准号: 7893161
• 负责人: VINCENT R RACANIELLO
• 金额: $ 40.68万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7893161
• 关键词: Address; Adult; Amino Acids; Animal Model; Animal Viruses; Antiviral Agents; Architecture; Asthma; Biology; CDPdiacylglycerol-inositol 3-phosphatidyltransferase; Cell Culture Techniques; Cells; Cleaved cell; Common Cold; Complex; Cultured Cells; Cytoplasm; DNA; DNA Sequence Rearrangement; Dendritic Cells; Dominant-Negative Mutation; Enzymes; Family; Genome; Goals; Golgi Apparatus; Growth; Human; Human poliovirus; Infection; Interferons; Internal Ribosome Entry Site; Laboratories; Lead; Life; Lung; Lytic; Mediating; Medical; Membrane; Molecular; Mus; Neonatal; Oral Poliovirus Vaccine; Organ; Pathogenesis; Pattern Recognition; Pattern recognition receptor; Play; Polioviruses; Primates; Protein Secretion; Proteins; RNA Viruses; Reporter; Research; Research Personnel; Respiratory System; Respiratory tract structure; Rhinovirus; Role; Signal Transduction; Site; Structure; Surface; TLR3 gene; TLR7 gene; Therapeutic Intervention; Transgenic Mice; Translations; Tropism; Vesicle; Viral; Viral Genome; Viral Proteins; Virus; Virus Diseases; attenuation; base; cytokine; design; human disease; inhibitor/antagonist; lipid biosynthesis; macromolecule; mouse model; mutant; pathogen; poliovirus receptor; prevent; programs; research study; response; sensor; trafficking

DESCRIPTION (provided by applicant): Human rhinoviruses (HRVs) are major causative agents of the common cold, one of the most frequent human infections. They are also the most common pathogens associated with asthma exacerbations. Despite the medical importance of rhinoviruses, the pathogenesis of rhinovirus infection is poorly understood because a convenient animal model is not available. The experiments in this proposal address a wide range of experimental questions on the biology of rhinovirus, encompassing replication in cell culture, the molecular basis of host range, innate responses to infection, and establishing replication in mice. These goals will be accomplished through the following four specific aims. 1. Determine the role in rhinovirus replication of cell proteins that interact with the viral 2B and 3A proteins. We have shown that the 3A protein of HRV39 interacts with GCP60 and FinGERS, proteins involved in vesicle traffic, and with an enzyme involved in lipid biosynthesis, CDIPT (phosphatidylinositol synthase). Cell proteins that interact with 2B will also be identified. We will assess the role of these cell proteins in viral replication, alteration of membrane ultrastructure, and inhibition of ER-to-Golgi traffic by reducing the levels of these proteins in the cell, interfering with their function with dominant negative forms of the proteins, or by producing non-interacting altered viruses. 2. Determine the role of the rhinovirus IRES in viral host range. We have found that the IRES of HRV2 does not mediate translation of a reporter protein in most organs of adult mice, including the lung. However, HRV2 IRES-mediated translation takes place in many organs of neonatal mice. Experiments in this specific aim are designed to determine whether the IRES of RV16, RV39, and RV1A can function in cells of the mouse lung. These cell cultures will be used to determine if specific structured regions of the IRES prevent internal initiation in primary lung cells from adult mice. Experiments are also planned to determine whether the lack of HRV2 IRES-mediated activity in cultured mouse cells is due to an inhibitor or absence of a required protein, and experiments to identify such factors are proposed. 3. Determine the role of pattern recognition molecules in rhinovirus replication. Experiments in this specific aim are designed to determine the role of specific pattern recognition molecules in rhinovirus replication. We will determine the roles of RIG-I, MDA-5, TLR3, and TLR7 in sensing rhinovirus replication in cultured cells. We have found that RIG-I and MDA-5 are cleaved in cells infected with rhinoviruses. Experiments are planned to determine whether cleavage of these sensor molecules benefits viral replication. In addition, we will determine whether rhinoviruses can replicate in dendritic cells, and whether TLR3 or TLR7 plays a role in sensing the viral genome in these cells. The identification of cell proteins that play a role in rhinovirus replication may reveal new targets for therapeutic intervention of the common cold.

描述（由申请方提供）：人鼻病毒（HRV）是普通感冒的主要病原体，普通感冒是最常见的人类感染之一。它们也是与哮喘急性发作相关的最常见病原体。尽管鼻病毒在医学上具有重要意义，但由于缺乏方便的动物模型，鼻病毒感染的发病机制仍知之甚少。本提案中的实验涉及鼻病毒生物学的广泛实验问题，包括细胞培养中的复制、宿主范围的分子基础、对感染的先天反应以及在小鼠中建立复制。这些目标将通过以下四个具体目标来实现。1.确定与病毒2B和3A蛋白相互作用的细胞蛋白在鼻病毒复制中的作用。我们已经表明，3A蛋白的HRV 39与GCP 60和FinGERS，参与囊泡交通的蛋白质，并与参与脂质生物合成，CDIPT（磷脂酰肌醇合酶）的酶相互作用。还将鉴定与2B相互作用的细胞蛋白。我们将评估这些细胞蛋白在病毒复制，膜超微结构的改变，并通过降低这些蛋白在细胞中的水平，干扰它们的功能与显性负形式的蛋白质，或通过生产非相互作用的改变病毒抑制ER到高尔基体交通的作用。2.确定鼻病毒IRES在病毒宿主范围内的作用。我们已经发现，在成年小鼠的大多数器官中，包括肺，HRV 2的IRES不介导报告蛋白的翻译。然而，HRV 2 IRES介导的翻译发生在新生小鼠的许多器官中。在此特定目的中的实验被设计成确定RV 16、RV 39和RV 1A的IRES是否可以在小鼠肺的细胞中起作用。这些细胞培养物将用于确定IRES的特定结构区域是否阻止成年小鼠原代肺细胞的内部启动。还计划进行实验以确定培养的小鼠细胞中缺乏HRV 2 IRES介导的活性是否是由于抑制剂或缺乏所需的蛋白质，并提出了鉴定此类因子的实验。3.确定模式识别分子在鼻病毒复制中的作用。在这一特定目标的实验设计，以确定特定的模式识别分子在鼻病毒复制的作用。我们将确定RIG-I，MDA-5，TLR 3和TLR 7在培养细胞中感知鼻病毒复制中的作用。我们已经发现RIG-I和MDA-5在被鼻病毒感染的细胞中被切割。计划进行实验以确定这些传感器分子的切割是否有利于病毒复制。此外，我们将确定鼻病毒是否可以在树突状细胞中复制，以及TLR 3或TLR 7是否在这些细胞中感知病毒基因组中发挥作用。对在鼻病毒复制中起作用的细胞蛋白的鉴定可能揭示普通感冒治疗干预的新靶点。