IMMUNOLOGICAL RESPONSE TO INFLUENZA INFECTION IN NH PRIMATES

NH 灵长类动物对流感感染的免疫反应

• 批准号: 8357600
• 负责人: MICHAEL G KATZE
• 金额: $ 37.79万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8357600
• 关键词: Acute; Animals; Attention; Blood; Cell Separation; Cells; Communicable Diseases; Communities; Flow Cytometry; Funding; Gene Expression; Gene Proteins; Genomics; Goals; Grant; Harvest; Immune; Immune response; Immunology; Infection; Influenza; Investigation; Laboratories; Lymph; Methods; Modeling; National Center for Research Resources; Performance; Population; Primates; Principal Investigator; Progress Reports; Proteins; Proteomics; RNA; Reporting; Research; Research Infrastructure; Resources; SIV; Sorting - Cell Movement; Source; Techniques; Time; United States National Institutes of Health; cell type; cost; functional genomics; influenzavirus; instrument; interest; nonhuman primate; response; virus host interaction

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The goal of this project is to use a nonhuman primate infection model, combined with genomic analyses of specific cell types isolated from the blood and lymph of infected animals, to define influenza virus-host interactions and the innate and adaptive immune responses to infection. During the current reporting period, we focused on establishing the techniques and parameters for sorting cells by flow cytometry (using a newly acquired FACS Aria instrument) and for optimizing the timing and methods for harvesting RNA and protein from isolated cells for subsequent analysis of cellular gene expression and protein abundance. Unfortunately, a large number of technical hurdles were uncovered in the effort to obtain suitable RNA from sorted cell populations. Despite repeated efforts, only low-quality RNA could be obtained from sorted cells. Our choice to examine an influenza infection model was largely premised on the interest of the Immunology Core to do a detailed characterization of sorted immune cell subsets. Other nonhuman primate influenza-infection models have already been generally characterized by the Katze Laboratory, including ongoing proteomics investigations with the Smith Laboratory. Consequently, the difficulty in obtaining sorted cells suitable for RNA isolation greatly diminished the potential scientific impact of this investigation. For this reason, we have chosen to focus attention on a different nonhuman primate model where the overall scientific questions are not so highly dependent on the performance of cell sorting. In consideration of unmet needs in the scientific community, we have redirected our resources to performing an acute SIV-infection study; details are given elsewhere in the progress report, under the general narrative for the Division of Functional Genomics and Infectious Disease.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 该项目的目的是使用非人类灵长类动物感染模型，并结合从感染动物的血液和淋巴中分离出的特定细胞类型的基因组分析，定义流感病毒 - 霍斯特 - 霍斯特 - 霍斯特 - 霍斯特 - 霍斯特相互作用以及对感染的先天和适应性免疫反应。在当前的报告期间，我们专注于通过流式细胞仪（使用新获得的FACS ARIA仪器）分类细胞的技术和参数，并优化从分离的细胞中收集RNA和蛋白质的时间和方法，以随后分析细胞基因表达和蛋白质的丰富度。不幸的是，在从分类的细胞群中获得合适的RNA的努力，发现了许多技术障碍。尽管一再进行了努力，但只能从分类的细胞中获得低质量的RNA。我们选择检查流感感染模型的选择基本上是基于免疫学核心的利益，以对分类的免疫细胞亚群进行详细表征。其他非人类灵长类动物感染模型通常以Katze实验室为特征，包括与Smith实验室进行的蛋白质组学研究。因此，获得适合RNA分离的分类细胞的困难大大减少了这项研究的潜在科学影响。因此，我们选择将注意力集中在不同的非人类灵长类动物模型上，因为整体科学问题不太依赖细胞分选的性能。考虑到科学界未满足的需求，我们将资源重定向到进行急性SIV感染研究。在进度报告中的其他地方给出了细节，该详细信息是在功能基因组学和传染病的一般叙述下。