Canine Influenza Virus Induction of TNF

犬流感病毒诱导TNF

• 批准号: 8355335
• 负责人: WILLIAM L CASTLEMAN
• 金额: $ 7.09万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8355335
• 关键词: Aerosols; Alveolar; Alveolar Macrophages; Animals; Antigens; Biological Assay; Bronchitis; CCL8 gene; Canis familiaris; Cells; Centers for Disease Control and Prevention (U.S.); Cessation of life; Collaborations; Coughing; Data; Disease; Distant; Eligibility Determination; Epithelial Cells; Equine Influenza Virus; Equus caballus; Feasibility Studies; Florida; Funding; Future; Gene Mutation; Genes; Goals; Grant; Hemorrhage; Human; IL8 gene; Immune response; In Vitro; Infection; Inflammatory Response; Influenza; Influenza A virus; Injury; Kentucky; Letters; Lung; Lung diseases; Maps; Measures; Messenger RNA; Methods; Modeling; Mutation; National Institute of Allergy and Infectious Disease; Pathogenesis; Plaque Assay; Pneumonia; Production; Protein Biosynthesis; Proteins; Pulmonary Edema; Race; Research; Research Project Grants; Ruthenium Ben; System; TNF gene; Time; Tissues; Tracheitis; Tumor Necrosis Factor-alpha; Universities; Veterinary Medicine; Viral; Virus; Virus Activation; Virus Diseases; Virus Replication; chemokine; college; cytokine; influenzavirus; macrophage; man; overexpression; positional cloning; response; transmission process; virus genetics

DESCRIPTION (provided by applicant): The long term goal of this research is to use H3N8 influenza virus as a model to determine viral genetic mutations in single or multiple gene segments of influenza virus A that facilitate interspecies transmission and induction of severe pulmonary disease. The hypothesis being pursued in these studies is: Transmission of equine influenza virus to dogs and adaptation in the new host to induce severe respiratory disease was characterized by mutations in one or several gene segments that allowed the virus to replicate to a greater extent in macrophages and induce higher levels of TNF-¿. The specific aims of this pilot research project are: 1)To compare virus replication and induction of TNF-¿ mRNA and protein synthesis and other cytokines in canine alveolar macrophages following inoculation with H3N8 canine influenza viruses from 2003-2004 with those following inoculation with H3N8 equine influenza virus isolates ranging from 2003 to 1991; 2) To determine whether the canine influenza virus derived with a reverse genetics system (canine/FL/04-rg) replicates to similar extent and induces comparable levels of TNF-¿ mRNA and protein in canine alveolar macrophages as wild type virus (A/canine/Florida/43/2004) does; and 3) To compare the capacity of canine and equine influenza virus isolates to replicate in primary airway epithelial cells and induce chemokines (IL-8 and MCP-2). The project will use primary isolated dog alveolar macrophages inoculated in vitro with equine influenza A viruses isolated from 1991 through 2003 and initial canine influenza isolates from 2003 and 2004. TNF-¿ mRNA and protein will be measured with quantitative real-time PCR and antigen capture assays, respectively. Virus production will be measured by infectious plaque assay and virus matrix gene quantitative real-time PCR. PUBLIC HEALTH RELEVANCE: The long term goal of this research is to understand which genetic changes in influenza virus from horses allowed it to be able to infect dogs and cause serious respiratory disease that could be passed on to other dogs. Equine influenza viruses from 1991 through 2003 will be compared with canine influenza virus first identified in 2003-2004 for their ability to induce a protein (tumor necrosis factor or TNF) when lung cells become infected with virus. Increased tumor necrosis factor secretion by lung cells is believed to cause severe changes in the lung that result in death. Information from this study will be used in other studies to identify the specific gene changes that allowed the virus to become deadly in dogs. The information will help better understand how influenza viruses from animals evolve to become infectious to other animals and man and cause disease.

描述（由申请人提供）：本研究的长期目标是使用H3 N8流感病毒作为模型，以确定甲型流感病毒单个或多个基因片段中的病毒基因突变，这些突变有助于种间传播和诱导严重肺部疾病。在这些研究中所追求的假设是：马流感病毒传播给狗和适应新的主机，以诱导严重的呼吸道疾病的特点是突变的一个或几个基因片段，使病毒复制到更大程度上在巨噬细胞和诱导更高水平的TNF-α。本试验项目的具体目的是：1）比较2003-2004年接种H3 N8犬流感病毒与2003 - 1991年接种H3 N8马流感病毒分离株后，犬肺泡巨噬细胞中病毒复制和TNF-α mRNA和蛋白质合成以及其他细胞因子的诱导; 2）确定犬流感病毒是否通过反向遗传学系统获得（犬/FL/04-rg）复制程度相似，并在犬肺泡巨噬细胞中诱导与野生型病毒相当的TNF-α mRNA和蛋白水平（A/canine/佛罗里达/43/2004）进行;和3）比较犬和马流感病毒分离物在原代气道上皮细胞中复制和诱导趋化因子（IL-8和MCP-2）的能力。该项目将使用1991年至2003年分离的马甲型流感病毒和2003年和2004年的初始犬流感病毒分离株体外接种的原代分离犬肺泡巨噬细胞。将分别采用定量实时PCR和抗原捕获试验测量TNF-α mRNA和蛋白质。将通过感染性空斑试验和病毒基质基因定量实时PCR测定病毒产量。 公共卫生相关性：这项研究的长期目标是了解马流感病毒的哪些遗传变化使其能够感染狗并导致严重的呼吸道疾病，这些疾病可能会传染给其他狗。1991年至2003年的马流感病毒将与2003年至2004年首次发现的犬流感病毒进行比较，以确定它们在肺细胞感染病毒时诱导蛋白质（肿瘤坏死因子或TNF）的能力。肺细胞的肿瘤坏死因子分泌增加被认为会导致肺的严重变化，从而导致死亡。这项研究的信息将用于其他研究，以确定允许病毒在狗中变得致命的特定基因变化。这些信息将有助于更好地了解来自动物的流感病毒如何演变成对其他动物和人类具有传染性并导致疾病。