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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.

项目摘要一个案件正在为实用程序的MeV持久性发展的终身免疫力。思想除免疫细胞外，上皮细胞的持续感染可产生病毒或病毒性免疫缺陷。抗原以促进抗体的亲和力成熟足够的持续时间，产生最高的亲合力。上皮细胞中表型的形成是由什么决定的是否有助于建立持续性感染尚不清楚。然而，这样的细胞应该能够以抑制但不消除病毒增殖，并且病毒释放必须通过非裂解性的避免慢性炎症状态的机制。在这种情况下，持续存在的- 感染感受态上皮细胞及其基因表达，特别是参与控制病毒增殖，是理解病毒持久性的分子基础的重要的第一步。我们对脊髓灰质炎病毒（PV）感染的单细胞分析显示，HeLa细胞群相当活跃，在感染后长达36小时不裂解的情况下，能够熟练地支持PV增殖，实验的持续时间。我们开发了一种策略来隔离这些持续性- 通过向培养物中加入PV进入抑制剂，使用标准细胞培养方法感染细胞媒体我们观察到PV持续时间长达6天感染后，再次的持续时间，实验对这些持续感染细胞的基因表达分析显示，抑制病毒和其他病原体的传感器，包括Toll样受体和“免费” （胞质）受体18. IL-32的基因被诱导7倍（P值<0.001）。IL-32已被与独立于I型和II型干扰素（IFN）的抗病毒应答有关;然而， IL-32依赖性抗病毒途径的效应物尚未发现。的首要目标我们将在R21基金的赞助下启动的项目是阐明基因，途径，和病毒持久性所需的机制。本申请的第一个目标是鉴定可能有助于建立CVB 3在人心肌细胞中的持久性。的第二个目标该应用激发了IL-32作用机制的假说。将努力实现这些目标本研究的主要目的是：（1）发现与CVB 3在人体内持续存在有关的基因（2）发现HeLa细胞中IL-32依赖性抗病毒状态的效应子。