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 人们正在论证 MeV 持久性在终生免疫力发展中的效用。 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 产生最高亲和力的因素决定了上皮细胞中表型的建立。 是否有利于建立持续感染尚不清楚，但是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。 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 我们制定了一项策略来隔离这个持续存在的亚群。 通过在培养物中添加 PV 进入抑制剂，使用标准细胞培养方法感染细胞 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。 对这些持续感染细胞的基因表达分析揭示了接近完整的结果。 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）。 然而，与 I 型和 II 型干扰素 (IFN) 无关的抗病毒反应有关； IL-32依赖性抗病毒途径的效应器尚未被发现。 我们将在 R21 拨款资助下启动的计划是阐明基因、途径、 以及病毒持久性所需的机制。该应用的第一个目标是识别基因。 可能有助于建立 CVB3 在人类心肌细胞中的持久性。 该应用旨在激发对 IL-32 作用机制的假设，以实现这些目标。 如下目标所示：（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。