Human cytomegalovirus latency in cultured monocytes

培养单核细胞中的人巨细胞病毒潜伏期

• 批准号: 8416378
• 负责人: THOMAS E SHENK
• 金额: $ 33.71万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-02-15 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8416378
• 关键词: Antiviral Therapy; Biological; Biological Assay; CD14 gene; Cell Culture Techniques; Cell surface; Cells; Clinical; Code; Cultured Cells; Cytomegalovirus; Cytomegalovirus Infections; Dendritic Cells; Genes; Genetic; Grant; Herpesviridae; Immune; Immune system; Individual; Infection; Investigation; Laboratories; Latent Virus; Maintenance; Messenger RNA; MicroRNAs; Modeling; Molecular; Molecular Biology; Monitor; Morbidity - disease rate; Myeloid Cells; Pharmaceutical Preparations; Phenotype; Play; Population; Proteomics; Role; Symptoms; System; Testing; Time; Viral; Viral Drug Resistance; Viral Genes; Viral Proteins; Virus; Virus Diseases; cell type; cytokine; deep sequencing; design; latent infection; macrophage; monocyte; mortality; mutant; peripheral blood; programs; public health relevance

DESCRIPTION (provided by applicant): Although they reside in different cell types, all herpes viruses can enter a state of quiescence termed latency. When these viruses are latent, they are resistant to antiviral therapies that attack replicating viruses. As a result, existing drugs treat symptoms of active infections, but do not clear latent virus, and, therefore, do not cure herpes virus infections. Consequently, it is critically important to advance our understanding of the mechanisms underlying latency; indeed, this is one of the major outstanding questions in our field. Human cytomegalovirus (HCMV) resides in a latent state in cells of the myeloid lineage. Although the virus likely reactivates frequently from latency, it is readily controlled with no known consequences to healthy people. However, reactivation can cause severe morbidity and mortality when it re-emerges from latency in people with compromised or immature immune systems. Thus, it is critical to develop the ability to control and eventually eradicate latent virus, and this capability will come from a deeper understanding of latency. My long term objective is to employ a validated cell culture model of latency -- peripheral blood monocytes -- to investigate the molecular mechanisms underlying the initiation, maintenance and exit from latency. We will carry out our investigations primarily with clinical isolates of HCMV; and our technical approach will combine genetics, molecular biology and proteomics. My specific aims are designed to employ our cell culture model of latency to (i) investigate the effect of cell phenotype on HCMV latency; (ii) study the role of viral protein-coding genes with the potential to function during latency; and (iii) test the hypothesis that viral and/or cell-coded miRNAs function in HCMV latency. This is an opportune time to study HCMV latency, because we now appreciate many of the critical differences between clinical versus laboratory isolates of the virus that cause them to behave differently in experimental infections, experimental systems have been developed to investigate latency in cultured cells, and new biological components likely to play important roles in latency have been recently identified.

描述（由申请人提供）：尽管它们居住在不同的细胞类型中，但所有疱疹病毒均可进入称为延迟的静止状态。当这些病毒潜在时，它们对攻击复制病毒的抗病毒疗法具有抵抗力。结果，现有药物治疗活性感染的症状，但不能清除潜在病毒，因此不能治愈疱疹病毒感染。因此，促进我们对潜伏潜在机制的理解至关重要。确实，这是我们领域的主要问题之一。人类巨细胞病毒（HCMV）位于髓样谱系细胞中的潜在状​​态。尽管该病毒可能经常从潜伏期中重新激活，但它很容易控制，对健康人没有已知后果。但是，当患有受损或不成熟的免疫系统患者潜伏期中重新出现潜伏期时，重新激活可能会导致严重的发病率和死亡率。因此，发展控制能力并最终消除潜在病毒至关重要，并且这种能力将来自对潜伏期的深刻理解。我的长期目标是采用经过验证的潜伏细胞培养模型（外周血单核细胞）来研究开始，维持和退出潜伏期的分子机制。我们将主要使用HCMV的临床分离株进行调查；我们的技术方法将结合遗传学，分子生物学和蛋白质组学。我的具体目的旨在利用我们的细胞培养模型的潜伏期来研究细胞表型对HCMV潜伏期的影响； （ii）研究病毒蛋白编码基因在潜伏期内的潜力的作用； （iii）检验以下假设：病毒和/或细胞编码的miRNA在HCMV潜伏期中的功能。这是研究HCMV潜伏期的时机，因为我们现在欣赏病毒的临床与实验室分离株之间的许多关键差异，这使它们在实验感染中的行为不同，已经开发了实验系统来研究培养细胞的潜伏期，而新的生物学成分可能在潜伏期中发挥了重要作用。