IMMUNOLOGICAL RESPONSE TO PRIMARY & SECONDARY INFLUENZA INFECTION IN NH PRIMATES

原发性免疫反应

• 批准号: 8172762
• 负责人: MICHAEL G KATZE
• 金额: $ 15.51万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8172762
• 关键词: Acute; Animals; Attention; Blood; Cell Separation; Cells; Communicable Diseases; Communities; Computer Retrieval of Information on Scientific Projects Database; Flow Cytometry; Funding; Gene Expression; Genomics; Goals; Grant; Harvest; Immune; Immune response; Immunology; Infection; Influenza; Institution; Investigation; Laboratories; Lymph; Methods; Modeling; Performance; Population; Primates; Progress Reports; Proteins; Proteomics; RNA; Reporting; Research; Research Personnel; Resources; Sorting - Cell Movement; Source; Techniques; Time; United States National Institutes of Health; cell type; 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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 该项目的目标是使用非人类灵长类动物感染模型，结合从受感染动物的血液和淋巴中分离的特定细胞类型的基因组分析，来定义流感病毒与宿主的相互作用以及对感染的先天和适应性免疫反应。在本报告期内，我们的重点是建立流式细胞仪分选细胞的技术和参数（使用新购置的 FACS Aria 仪器），以及优化从分离细胞中收获 RNA 和蛋白质的时间和方法，以便随后分析细胞基因表达和蛋白质丰度。不幸的是，在从分选的细胞群中获取合适的 RNA 的过程中发现了大量的技术障碍。尽管经过多次努力，仍只能从分选的细胞中获得低质量的RNA。我们选择检查流感感染模型很大程度上是基于免疫学核心对分选的免疫细胞子集进行详细表征的兴趣。卡茨实验室已经对其他非人类灵长类流感感染模型进行了总体表征，包括史密斯实验室正在进行的蛋白质组学研究。因此，获得适合 RNA 分离的分选细胞的困难大大削弱了这项研究的潜在科学影响。出于这个原因，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，其中整体科学问题并不那么高度依赖于细胞分选的性能。考虑到科学界未满足的需求，我们已将资源重新用于开展急性 SIV 感染研究；进展报告中功能基因组学和传染病部门的总体叙述中提供了详细信息。