Contribution of IL-32 gene expression to viral persistence

IL-32 基因表达对病毒持久性的贡献

• 批准号: 10057016
• 负责人: CRAIG E. CAMERON
• 金额: $ 24.24万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-03 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057016
• 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

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 正在为MEV持久性在终身免疫力发展方面的效用而提出案例。想法 除了免疫细胞外，上皮细胞的持续感染还可能产生病毒或病毒 抗原促进抗体的亲和力成熟，以在足够的持续时间内 产生最高的亲和力。是什么控制上皮细胞中表型的建立 建立持续感染的导电尚不清楚。但是，这样的单元应该能够 要抑制但不能消除病毒繁殖，并且必须通过非散型发生病毒释放 避免慢性炎症状态的机制。建立这种持久的存在 - 感染竞争性上皮细胞及其基因表达，尤其是与控制的基因 病毒繁殖是理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）感染的单细胞分析表明，HELA细胞群体非常 感染后36小时，熟练支持PV繁殖，而无需裂解，这是 实验的持续时间。我们制定了隔离这种持续的亚群的策略 - 通过标准细胞培养方法感染细胞，通过将PV进入的抑制剂添加到培养物中 媒体。我们在感染后六天观察到PV持久性，再次持续 实验。对这些持续感染的细胞的基因表达分析显示出一个近乎完整的 抑制病毒和其他病原体的传感器，包括收费受体和“无电话” （胞质）受体18。诱导IL-32的基因7倍（p值<0.001）。 IL-32已经 在独立于I型和II型干扰素（IFN）的抗病毒反应中实施；然而， IL-32依赖性，抗病毒途径的效应因子尚未发现。总体目标 我们将在此R21赠款的主持下启动的程序是阐明基因，途径， 病毒持久性所需的机制。该应用程序的第一个目标是确定基因 可能有助于建立人类心肌细胞中CVB3的持久性。第二个目标 该应用是为了激发IL-32动作机制的假设。这些目标将被追求 如以下目的所示：（1）发现有助于人类CVB3持续存在的基因 心肌细胞； （2）发现IL-32依赖性抗病毒态在HELA细胞中的影响。