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Contribution of IL-32 gene expression to viral persistence

IL-32 基因表达对病毒持久性的贡献

基本信息

批准号：
10177863
负责人：
CRAIG E. CAMERON
金额：
$ 18.95万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-03 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10177863
关键词：
Affinity Antibodies Antiviral Agents Antiviral Response Avidity Biomedical Engineering Cardiac Myocytes Cardiomyopathies Cell Culture Techniques Cell Line Cell Nucleus Cells Chronic Collaborations Complementary DNA Coxsackie Viruses Coxsackievirus Infections Culture Media Cytolysis Cytoplasm Development Epithelial Cells Gene Expression Gene Expression Profiling Genes Genotype Goals Grant Hela Cells Human Human poliovirus Immune Immunity Infection Inflammation Innate Immune Response Interferon-alpha Interferon-beta Interferons Knock-out Knowledge Measles virus Mediator of activation protein Modeling Molecular Nonlytic Pathway interactions Pharmacology Phenotype Population Predisposition Protein Isoforms Proteomics Public Health RNA Viruses Regimen System Therapeutic Time Toll-like receptors Up-Regulation Viral Viral Antigens Virus Virus Diseases Virus Replication Work acute infection base chronic infection experimental study gene discovery induced pluripotent stem cell inhibitor/antagonist pathogen personalized medicine programs prophylactic response sensor single cell analysis transcriptome

项目摘要

Project Abstract A case is being made for the utility of MeV persistence in development of lifelong immunity. The thought is that persistent infection of epithelial cells, in addition to immune cells, may produce virus or viral antigens to promote affinity maturation of antibodies for a sufficient duration of time that antibodies of the highest avidity are produced. What governs establishment of a phenotype in an epithelial cell conducive to establishment of a persistent infection is not known. However, such a cell should be able to suppress but not eliminate virus multiplication, and virus release must occur by a non-lytic mechanism to avoid a state of chronic inflammation. Establishing the existence of such persistent- infection-competent epithelial cells and their gene expression, especially genes involved in control of virus multiplication, is an essential first step in understanding the molecular basis of viral persistence. Our single-cell analysis of poliovirus (PV) infection revealed a population of HeLa cells that were quite proficient at supporting PV multiplication without lysis for as long as 36 h post-infection, which was the duration of the experiment. We developed a strategy to isolate this sub-population of persistently- infected cells using standard cell-culture approaches by adding an inhibitor of PV entry to the culture media. We observed PV persistence for as long as six days post-infection, again the duration of the experiment. Gene-expression analysis of these persistently-infected cells revealed a near-complete suppression of sensors of viruses and other pathogens, including Toll-like receptors and “toll-free” (cytosolic) receptors18. The gene for IL-32 was induced 7 fold (P-value <0.001). IL-32 has been implicated in antiviral responses that are independent of type I and II interferons (IFNs); however, effectors of the IL-32-dependent, antiviral pathway have yet to be discovered. The overarching goal of the program that we will initiate under the auspices of this R21 grant is elucidation of genes, pathways, and mechanisms required for viral persistence. The first goal of this application is to identify genes that may contribute to establishment of persistence of CVB3 in human cardiomyocytes. The second goal of this application is to inspire hypotheses for the mechanism of IL-32 action. These goals will be pursued as indicated by the following aims: (1) Discover genes contributing to CVB3 persistence in human cardiomyocytes; and (2) Discover effectors of the IL-32-dependent antiviral state in HeLa cells.

项目摘要人们正在论证 MeV 持久性在终生免疫力发展中的效用。这个想法上皮细胞的持续感染，除了免疫细胞外，还可能产生病毒或病毒抗原以促进抗体的亲和力成熟足够长的时间，使抗体产生最高的亲和力。上皮细胞表型的建立受什么控制是否有助于建立持续感染尚不清楚。然而，这样的细胞应该能够抑制但不能消除病毒增殖，并且病毒释放必须通过非裂解性方式发生避免慢性炎症状态的机制。确定这种持久性的存在有感染能力的上皮细胞及其基因表达，特别是参与控制感染的基因病毒增殖是了解病毒持久性分子基础的重要第一步。我们对脊髓灰质炎病毒 (PV) 感染的单细胞分析揭示了一群 HeLa 细胞，它们与感染后长达 36 小时，能够熟练地支持 PV 增殖而无需裂解，这是实验的持续时间。我们制定了一项策略来隔离这个持续存在的亚群使用标准细胞培养方法通过在培养物中添加 PV 进入抑制剂来感染细胞媒体。我们观察到感染后 PV 持续存在长达六天，这也是感染的持续时间。实验。这些持续感染细胞的基因表达分析揭示了近乎完整的抑制病毒和其他病原体的传感器，包括 Toll 样受体和“toll-free” （胞质）受体18。 IL-32 基因被诱导7倍（P值<0.001）。 IL-32 已参与独立于 I 型和 II 型干扰素 (IFN) 的抗病毒反应；然而， IL-32 依赖性抗病毒途径的效应器尚未被发现。总体目标是我们将在 R21 拨款资助下启动的计划是阐明基因、途径、以及病毒持续存在所需的机制。该应用程序的第一个目标是识别基因可能有助于建立人类心肌细胞中 CVB3 的持久性。第二个目标是该应用旨在激发对 IL-32 作用机制的假设。这些目标将被追求其目标如下：（1）发现有助于人类CVB3持久性的基因心肌细胞； (2) 发现 HeLa 细胞中 IL-32 依赖性抗病毒状态的效应器。