VIROLOGY CORE A

项目摘要(见说明)：呼吸道病毒感染，特别是人鼻病毒(HRV)感染，在哮喘的发生、发展、恶化和慢性化方面发挥着重要作用。该计划的重点是确定调节HRV复制的因素和宿主对病毒感染的免疫反应之间的相互作用，这些因素影响相关呼吸道疾病的严重程度。每个项目都将使用病毒诊断，项目II和111需要专门制备纯化病毒和与病毒生物学有关的分子试剂。建立病毒学核心是为了向三个拟议项目中的每一个项目提供病毒诊断和HRV分子试剂。项目I将得到一系列灵敏和准确的分子检测分析的支持，以识别所有常见的呼吸道病毒和HRV的个别物种/类型/毒株。病毒载量将在项目I的人类受试者的HRV阳性鼻样的子集中通过qPCR进行量化，以用于后续的病毒分离、完整的基因组测序、cDNA克隆和在项目11-11中使用。此外，该中心还将支持L项目的体外实验，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(天然、放射性标记和灭活)，100个经典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)选定的HRV-A、HRV-B和HRV-C型/毒株的cDNA克隆(感染性、嵌合体、报告基因、复制子)，(4)进行感染性分析(空斑和TCID50)，(5)呼吸道上皮细胞的培养

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