Fate of neurons following immune-mediated viral clearance

免疫介导的病毒清除后神经元的命运

• 批准号: 8191430
• 负责人: GLENN F RALL
• 金额: $ 26.1万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8191430
• 关键词: Address; Age; Apoptotic; Brain; Cell Survival; Cells; Central Nervous System Viral Diseases; Cessation of life; Chronic; Color; Data; Embryo; Etiology; Event; Fibroblasts; Fluorescence; Gene Expression; Genes; Genetic Transcription; Heterogeneity; Histologic; Human; Immune; Immune response; Immunity; Immunocompetent; Impairment; In Situ Nick-End Labeling; In Vitro; Infection; Inflammatory; Knowledge; Link; Measles; Measles virus; Mediating; Modeling; Molecular Profiling; Monitor; Mus; Neurodegenerative Disorders; Neurons; Neuropathogenesis; Neurotransmitters; Neurovirology; Outcome; Phenotype; Play; Proteins; RNA Viruses; Rabies; Rabies virus; Recombinants; Relative (related person); Reporter; Role; Signal Transduction; Staining method; Stains; Synapses; System; Testing; Therapeutic; Time; Viral; Viral Antigens; Viral Vector; Virus; Virus Diseases; Work; base; cohort; congenic; flexibility; gene therapy; in vivo; laser capture microdissection; mouse model; neuronal survival; neurotropic; neurotropic virus; novel; pathogen; prevent; protein expression; receptor; recombinase; research study; response; viral RNA

DESCRIPTION (provided by applicant): Despite the large number of viruses that can infect the human brain, and the growing suspicion that some neurotropic pathogens play a direct or contributory role in chronic neurodegenerative diseases, little is known about the fate of neurons following infection. It has been surmised that neurons can survive an infection and immunity event, resulting in cured cells that are functionally indistinguishable from the pre-infection phonotype, though this has not been experimentally tested. While there is abundant evidence that immunocompetent mice mount a protective response within the CNS that resolves a viral infection, leading to little overt inflammatory damage and apparent impairment-free survival, two critical questions remain unanswered, which are the basis for this proposal. First: do neurons, in fact, survive the anti- viral immune response? Second: If neurons can be cured of a virus, are they functionally competent? These questions will be addressed using a well-characterized mouse system (congenic Cre reporter mice) to follow the consequences of viral infection of neurons over time. Neurons that have been infected with one of two neurotropic RNA viruses (measles or rabies) will be "marked" by a permanent change in fluorescence signal from red to green. These neurons can then be monitored throughout the infection course in the presence or absence of a competent immune response. Moreover, marked (green) neurons can be isolated by laser capture microdissection for subsequent gene and protein expression analyses. The ability to identify, isolate and characterize actively infected neurons or previously infected neurons that have been cured by the host immune response will allow for more precise studies to define how variables including age, neuronal subtype, virus type, and immune effectors impact on CNS neuron survival and function. Finally, given that this system can be broadly applied to virtually any mouse model of CNS viral infection, we believe there is the potential to enable major advances in the fields of neurovirology and neuropathogenesis. PUBLIC HEALTH RELEVANCE: This proposal will explore the utility of a novel mouse model to explore neuronal survival and function following virus infection. In this model, neurons that have been infected by Cre- recombinase expressing viruses undergo a permanent, readily detectable color change from red to green, serving as a marker of previous infection. Using two well-characterized neurotropic viruses (measles and rabies), neuronal survival and function will be assessed during active infection and at various timepoints post-clearance. The information obtained from these studies will have direct translational links that will guide therapeutic strategies to prevent or resolve neurotropic infections in humans, aid viral vector-based gene therapy strategies, and elucidate what role, if any, viruses play in neurodegenerative diseases of unknown etiology.

描述(申请人提供)：尽管有大量的病毒可以感染人脑，而且越来越多的人怀疑一些嗜神经的病原体在慢性神经退行性疾病中发挥了直接或促成作用，但人们对感染后神经元的命运知之甚少。据推测，神经元可以在感染和免疫事件中幸存下来，导致治愈的细胞在功能上与感染前的音型无法区分，尽管这还没有经过实验测试。虽然有大量证据表明，具有免疫能力的小鼠在中枢神经系统内发起保护性反应，解决病毒感染，导致几乎没有明显的炎症损伤和明显的无损伤存活，但有两个关键问题仍然没有得到回答，这两个问题是这一提议的基础。第一：神经元真的能在抗病毒免疫反应中存活下来吗？第二：如果神经元可以从病毒中治愈，它们的功能是否正常？这些问题将通过一个特征良好的小鼠系统(同源Cre报告小鼠)来解决，以跟踪随着时间的推移病毒感染神经元的后果。被两种嗜神经性RNA病毒(麻疹或狂犬病)中的一种感染的神经元将被永久的荧光信号从红色改变为绿色。然后，这些神经元可以在感染过程中进行监测，无论是否存在有效的免疫反应。此外，标记的(绿色)神经元可以通过激光捕获显微解剖分离出来，用于后续的基因和蛋白质表达分析。能够识别、分离和表征被宿主免疫反应治愈的活跃感染的神经元或先前感染的神经元，将允许更精确的研究，以确定包括年龄、神经元亚型、病毒类型和免疫效应器在内的变量如何影响中枢神经系统神经元的存活和功能。最后，鉴于该系统可以广泛应用于几乎任何中枢神经系统病毒感染的小鼠模型，我们相信有可能在神经病毒学和神经发病机制领域取得重大进展。 公共卫生相关性：这项提案将探索一种新的小鼠模型的用途，以探索病毒感染后神经元的存活和功能。在这个模型中，被表达Cre重组酶病毒感染的神经元经历了永久的、容易检测到的颜色变化，从红色到绿色，作为以前感染的标志。使用两种特性良好的嗜神经性病毒(麻疹和狂犬病)，将评估活跃感染期间和清除后不同时间点的神经元存活和功能。从这些研究中获得的信息将具有直接的翻译联系，将指导预防或解决人类嗜神经性感染的治疗策略，帮助基于病毒载体的基因治疗策略，并阐明病毒在不明原因的神经退行性疾病中扮演的角色(如果有的话)。