VIROLOGY CORE A

病毒学核心A

• 批准号: 8790529
• 负责人: Yury A Bochkov
• 金额: $ 24.08万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8790529
• 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

项目摘要（参见说明）：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、恶化和慢性化中发挥重要作用。该计划的重点是确定调节 HRV 复制的因素与宿主对病毒感染的免疫反应之间的相互作用，从而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相关的分子试剂。病毒学核心的建立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 将得到一系列灵敏且准确的分子检测分析的支持，以识别所有常见呼吸道病毒和 HRV 的单个物种/类型/菌株。将通过 qPCR 对项目 I 人类受试者的 HRV 阳性鼻腔样本子集中的病毒载量进行定量，以便随后进行病毒分离、完整基因组测序、cDNA 克隆并在项目 II-III 中使用。此外，该核心将通过提供（1）高滴度和统一的HRV病毒库存，包括选定的HRV血清型和毒株的纯化病毒（天然的、放射性标记的和灭活的）、100种经典HRV-A和HRV-B血清型的感染细胞裂解物以及新鉴定的HRV-C毒株，（2）各种抗HRV，支持L-III项目的体外实验。 抗体，(3) 所选 HRV-A、HRV-B 和 HRV-C 类型/株的 cDNA 克隆（感染性、嵌合性、报告基因、复制子），(4) 进行感染性测定（噬菌斑和 TCID50），以及 (5) 气道上皮细胞培养