Varicella-zoster Virus: Tegument Proteins in Pathogenesis

水痘带状疱疹病毒：发病机制中的皮层蛋白

• 批准号: 8121089
• 负责人: Ann Arvin
• 金额: $ 7.4万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-16 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8121089
• 关键词: Address; Adult; Afferent Neurons; Animal Model; Apoptosis; Attenuated; Autopsy; Binding; Binding Sites; Cell Culture Techniques; Cells; Chemical Agents; Chickenpox; Child; Disease; Elderly; Elements; Ensure; Equilibrium; Evaluation; Event; Exhibits; Ganglia; Generations; Genes; Genetic Transcription; Genome; Glycoproteins; Goals; Grant; Heating; Herpes zoster disease; Herpesviridae; Herpesvirus Type 3; Histone Deacetylase Inhibitor; Human; Immune; Immune system; In Vitro; Infection; Inhibition of Apoptosis; Interferons; Licensing; Life; Lytic Phase; Maps; Mediating; Membrane Proteins; Methods; Modeling; Molecular; Morbidity - disease rate; Mus; Mutate; Mutation; Myxoid cyst; Nerve Growth Factors; Neuroglia; Neurons; Neuropathogenesis; Neurotropism; Nuclear; Nuclear Envelope; Pathogenesis; Peripheral; Phosphorylation; Phosphotransferases; Population; Process; Production; Progress Reports; Protein Kinase; Proteins; Public Health; Recombinant Vaccines; Recombinants; Regulation; Reporter; Role; Sensory Ganglia; Severe Combined Immunodeficiency; Signal Transduction; Simplexvirus; Site; Skin; Spinal Ganglia; Staging; Stimulus; Study models; T-Lymphocyte; Testing; Time; Tissues; Trans-Activators; Transcript; Tropism; United States; Vaccines; Variant; Viral; Viral Genes; Viral Proteins; Virion; Virulence; Virus; Virus Diseases; Virus Replication; Work; Xenograft procedure; design; genetic regulatory protein; high risk; improved; in vivo; in vivo Model; mutant; neuron apoptosis; neurovirulence; prevent; promoter; protein expression; reactivation from latency; research study; response; satellite cell; skin xenograft; stressor; success; transcription factor; viral DNA; virus pathogenesis

Varicella zoster virus (VZV) causes varicella during primary infection, persists in sensory ganglia and may reactivate from latency to cause zoster. VZV pathogenesis depends upon its tropisms for T cells, skin and sensory ganglia. VZV vaccines to prevent varicella and to reduce zoster morbidity in the elderly are very effective. However, VZV control could be improved with a 2nd generation recombinant vaccine that has attenuated replication in skin, but limited infectivity for T cells and ganglia. VZV infection of sensory ganglia ensures its survival in the human population through reactivations from latency that result in zoster. We have developed a model for studying VZV neuropathogenesis by making xenografts of human dorsal root ganglia (DRG) in mice with severe combined immunodeficiency (SCID). Our analyses of VZV tegument/regulatory proteins will be extended to examine their functions in VZV neurotropism. We will focus on IE63, an important immediate early regulatory protein encoded by ORF63, and the ORF66 kinase protein using VZV recombinants that have targeted mutations in these genes. The SCIDhu DRG model also offers unique opportunities to investigate how cell transactivators that modulate viral gene promoters may control VZV neurotropism and to identify what perturbations of persistently infected neurons may trigger of VZV reactivation. The roles of IE63 and ORF66 at early and late stages of VZV infection of ganglia will be examined in SCIDhu DRG. Experiments will address four general hypotheses: 1) initial VZV replication is required for, or influences the level of persistent VZV DNA copies in neurons; 2) VZ virions must be made and released efficiently; 3) VZV proteins that inhibit neural cell apoptosis facilitate persistence; 4) VZV infection of DRG reflects an equilibrium with innate cellular responses, mediated by interferons (IFN), which optimizes persistence. VZV gene promoters, like those of all herpesviruses, have elements that are recognized by ubiquitous host cell regulatory proteins. Our hypothesis is that cellular proteins regulate transcription from the critical ORF63 promoter during initial infection and are needed for the transition to latency in sensory neurons. Experiments to understand what cellular stressors may induce VZV reactivation from persistently infected DRG will examine heat, chemical agents, histone deacetylase inhibitors, interference with nerve growth factor signaling and other triggers known to enhance herpes simplex virus reactivation. The work proposed for Yr. 1 and Yr. 2 (see Specific Aims) is expected to yield new information about the molecular mechanisms of VZV neuropathogenesis in differentiated peripheral neurons and satellite cells within human sensory ganglia and to identify options for designing a 2nd generation VZV vaccine with genetic changes that have been proved to reduce virulence in the SCIDhu DRG model in vivo.

水痘带状疱疹病毒（VZV）在原发感染期间引起水痘，持续存在于感觉神经节， 可能从潜伏期重新激活导致带状疱疹。VZV的发病机制取决于其对T细胞、皮肤 和感觉神经节。VZV疫苗，以防止水痘和减少带状疱疹发病率在老年人是 非常有效.然而，VZV控制可以通过第二代重组疫苗得到改善， 减弱皮肤中的复制，但对T细胞和神经节的感染性有限。感觉神经节VZV感染 通过从导致带状疱疹的潜伏期重新激活来确保其在人群中的存活。我们 通过制作人背根异种移植物，开发了一种研究VZV神经发病机制的模型 在患有严重联合免疫缺陷（SCID）的小鼠中的神经节（DRG）。我们对VZV的分析 被膜/调节蛋白将被扩展以检查它们在VZV嗜神经性中的功能。我们将重点 对IE63，一种由ORF 63编码的重要的立即早期调节蛋白，和ORF 66激酶 使用在这些基因中具有靶向突变的VZV重组体的蛋白质。SCIDhu DRG模型还 提供了独特的机会来研究细胞反式激活因子如何调节病毒基因启动子， 控制VZV嗜神经性，并确定持续感染的神经元的扰动可能引发 VZV重新激活。IE63和ORF 66在VZV感染神经节的早期和晚期阶段的作用将是 在SCIDhu DRG中检查。实验将解决四个一般假设：1）初始VZV复制是 需要或影响神经元中持续VZV DNA拷贝的水平; 2）VZ病毒体必须被制造 3）VZV蛋白抑制神经细胞凋亡，促进VZV的持久性; DRG的感染反映了与干扰素（IFN）介导的先天性细胞应答的平衡， 优化持久性。VZV基因启动子，像所有疱疹病毒的启动子一样， 被普遍存在的宿主细胞调节蛋白识别。我们的假设是细胞蛋白调节 在初始感染期间从关键的ORF 63启动子转录，并且是过渡到 感觉神经元的潜伏期。了解哪些细胞应激源可能诱导VZV再激活的实验 从持续感染的DRG将检查热，化学试剂，组蛋白脱乙酰酶抑制剂， 干扰神经生长因子信号传导和其他已知增强单纯疱疹病毒的触发因素 重新激活该工作提出了YR。1，Y。2（见具体目标）预计将产生新的信息 关于VZV在分化的外周神经元中神经发病的分子机制， 人类感觉神经节内的卫星细胞，并确定设计第二代VZV的选择 具有已被证明在体内SCIDhu DRG模型中降低毒力的遗传改变的疫苗。