VIROLOGY CORE A

病毒学核心 A

• 批准号: 8470004
• 负责人: Yury A Bochkov
• 金额: $ 19.05万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8470004
• 关键词: Acute; Adherent Culture; Adult; Affect; Antibodies; Asthma; Biological Assay; Biology; Cell Line; Cells; Child; Clinical; Cloning; Collection; Complementary DNA; Consult; Consultations; Detection; Development; Diagnostic; Epithelial Cells; Gene Proteins; Generations; Growth; Hela Cells; Hospitalization; Human; Human Resources; Immune response; In Vitro; Individual; Infant; Infection; Instruction; Lead; Modeling; Molecular; Molecular Biology; Nose; Organ Culture Techniques; Plaque Assay; Play; Preparation; Process; Production; Radiolabeled; Reagent; Replicon; Reporter; Research; Respiratory Tract Infections; Reverse Transcriptase Polymerase Chain Reaction; Rhinovirus; Role; Sampling; Serotyping; Severities; Severity of illness; Sinus; Specimen; Viral; Viral Load result; Virus; Virus Diseases; Wheezing; airway obstruction; genome sequencing; human subject; programs; radiotracer; recombinant virus; research study; respiratory; respiratory virus; viral RNA; virology

PROJECT SUMMARY (See instructions): Respiratory viral infections, particularly by human rhinoviruses (HRV), play important roles in inception, development, exacerbation and chronicity of asthma. This program is focused on defining interactions between factors regulating HRV replication and host immune response to viral infection that affect the severity of associated respiratory illness. Each of the projects will use viral diagnostics, and Projects II and 111 require specialized preparations of purified viruses and molecular reagents related to viral biology. The Virology Core is established to provide viral diagnostics and HRV molecular reagents to each of the three proposed projects. Project I will be supported with a repertoire of sensitive and accurate molecular detection assays to identify all common respiratory viruses and the individual species/type/strain of HRV. The viral loads will be quantified by qPCR in a subset of HRV-positive nasal samples from human subjects of Project I for subsequent virus isolation, complete genome sequencing, cDNA cloning and use in Projects ll-lll. In addition, this core will support the in vitro experiments of Projects l-lll by supplying (1) high-titer and uniform HRV virus stocks, including purified viruses (native, radiolabeled and inactivated) of selected HRV serotypes and strains, infected cell lysates of 100 classical HRV-A and HRV-B serotypes and newly identified HRV-C strains, (2) various anti-HRV antibodies, (3) cDNA clones (infectious, chimeric, reporter, replicons) of selected HRV-A, HRV-B and HRV-C types/strains, (4) performing infectivity assays (plaque and TCID50), and (5) culture of airway epithelial cells.

项目概述（见说明书）：呼吸道病毒感染，特别是人鼻病毒（HRV）感染，在哮喘的发病、发展、恶化和慢性化中起着重要作用。这个项目的重点是确定调节HRV复制的因素和宿主对影响相关呼吸系统疾病严重程度的病毒感染的免疫反应之间的相互作用。每个项目都将使用病毒诊断，项目II和项目111需要专门制备纯化病毒和与病毒生物学相关的分子试剂。建立病毒学核心是为了向三个拟议项目提供病毒诊断和HRV分子试剂。项目一将得到一系列敏感和准确的分子检测分析的支持，以确定所有常见的呼吸道病毒和HRV的单个物种/类型/毒株。在项目1的人类受试者的一部分hrv阳性鼻样本中，将通过qPCR定量检测病毒载量，用于后续的病毒分离、全基因组测序、cDNA克隆和项目2 - 3中使用。此外，该核心将通过提供(1)高滴度和统一的HRV病毒库，包括选定HRV血清型和毒株的纯化病毒（原生、放射性标记和灭活），100种经典HRV- a和HRV- b血清型和新鉴定的HRV- c毒株的感染细胞裂解物，(2)各种抗HRV抗体，(3)选定HRV- a、HRV- b和HRV- c型/毒株的cDNA克隆（感染性、嵌合性、报告性、复制子），支持项目l-ll的体外实验。(4)进行感染性实验（斑块和TCID50），(5)气道上皮细胞培养。