VZV in the enteric nervous system: pathogenesis and consequences

肠神经系统中的水痘带状疱疹病毒：发病机制和后果

• 批准号: 8153660
• 负责人: MICHAEL D GERSHON
• 金额: $ 40万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8153660
• 关键词: Afferent Neurons; Animal Model; Animals; Axon; Axonal Transport; Behavior Control; Blood; Brain; Breathing; Cavia; Cells; Chickenpox; Chickenpox Vaccine; Chronic; Coculture Techniques; Complex; Cranial Nerves; Cutaneous; Differential Diagnosis; Disease; Dorsal; Enteral; Enteric Nervous System; Enterocytes; Epithelium; Exanthema; Exocytosis; Fibroblasts; Frequencies; Functional disorder; Ganglia; Gastrointestinal Motility; Gastroparesis; Growth; Herpes zoster disease; Herpesvirus Type 3; Human; Immunity; Immunocompromised Host; Immunologic Deficiency Syndromes; In Situ; In Vitro; Individual; Infection; Infection prevention; Inflammation; Inflammatory Bowel Diseases; Injection of therapeutic agent; Intestinal Pseudo-Obstruction; Intestines; Investigation; Irritable Bowel Syndrome; Label; Life; Link; Lymphocyte; Lytic; Lytic Phase; Mus; Nerve; Nerve Block; Nerve Fibers; Neurons; Open Reading Frames; Operative Surgical Procedures; Paint; Pathogenesis; Peripheral Blood Mononuclear Cell; Phase; Proteins; Reverse Transcriptase Polymerase Chain Reaction; Route; Sensory Nerve Endings; Severities; Simplexvirus; Skin; Spinal Cord; Sympathectomy; Synapses; T-Lymphocyte; Testing; Tracer; Travel; Vesicle; Viral; Viral Proteins; Viremia; Virion; Virus; Virus Diseases; Virus Latency; Viscera; afferent nerve; base; design; experience; immunocytochemistry; in vitro Model; in vivo; intradermal injection; keratinocyte; killings; late endosome; latent infection; man; mutant; particle; preference; protein expression; research study; secondary infection; spinal nerve posterior root; true blue; viral DNA; visceral afferent nerve

DESCRIPTION (provided by applicant): The enteric nervous system (ENS) can control the behavior of the bowel without input from brain or spinal cord. A functioning ENS is essential for life and, when abnormal, causes discomfort and may contribute to the pathophysiology or severity of disorders of gastrointestinal motility, secretion, and inflammation. We have recently discovered that varicella zoster virus (VZV) establishes latency within human enteric neurons in most individuals who have experienced natural varicella or received varicella vaccine. VZV, moreover, has been linked to the occurrence of lethal pseudoobstruction in immunocompromised individuals. Neither the route by which VZV gains access to the ENS, nor the frequency or consequences of its reactivation in enteric neurons ("enteric zoster") has previously been explored. The current proposal is designed to test the hypotheses that transport in visceral afferent nerves conducts VZV to the ENS, that cell- free virions establish latency in enteric neurons, and that the non-structural VZV ORF61 protein must be expressed in neurons to enable VZV to manifest lytic infection or to reactivate from latency. Although VZV displays a marked preference for human cells, we have developed animal models that permit VZV infection of the ENS to be studied in vitro and in vivo. Depending on conditions, VZV recapitulates latent, lytic, and reactivating infection in enteric neurons isolated from guinea pigs or mice and, when introduced to the bowel, VZV establishes latency in the guinea pig ENS in situ. The proposal has 3 specific aims: (1) Can VZV travel from the skin to the ENS in sensory nerve fibers? Preliminary studies have identified neurons in dorsal route ganglia that project both to skin and gut. We will determine whether latent infection is established in the ENS when VZV is introduced to the skin and whether VZV-infected nerve terminals release infectious cell-free VZV that crosses synaptic gaps to transfer latent infection to target neurons. (2) Can a viremia establish latent VZV infection of enteric neurons? We will determine whether VZV-infected T lymphocytes release infectious cell-free VZV and whether they can establish latency directly in enteric neurons or indirectly via infections of the mucosal epithelium or skin. Preliminary studies have shown that VZV DNA is present in guinea pig enteric neurons following the iv injection of VZV-infected peripheral blood mononuclear cells. (3) Is VZV ORF61 protein expression necessary for the manifestation of lytic infection in enteric neurons? We will study the ability of a VZV mutant that lacks ORF61 to establish lytic infection of enteric neurons or reactivate from latency. The significance of understanding VZV infection of the ENS is enhanced by the possibility that unsuspected reactivation of VZV in enteric neurons might contribute to the pathogenesis of GI disorders such as irritable bowel syndrome, inflammatory bowel disease, idiopathic gastroparesis, and chronic intestinal pseudoobstruction. PUBLIC HEALTH RELEVANCE: The gut contains a large and complex network of nerve cells, known as the enteric nervous system (ENS), which is able to control the behavior of the bowel without input from the brain or spinal cord. The ENS contributes to the underlying basis of a number of disorders that disturb the functioning of ENS, such as irritable bowel syndrome, inflammatory bowel disease, idiopathic gastroparesis, and chronic intestinal pseudoobstruction are currently not understood. We have recently discovered that varicella zoster virus (VZV) establishes latency within nerve cells of the human ENS. Neither the cause nor the consequences of this phenomenon have previously been explored because latent VZV in enteric nerve cells was not known to occur. The current proposal is designed to determine how VZV gains access to the ENS and we will utilize enteric nerve cells to test the hypotheses that only the cell-free particle form of VZV is able to establish of latency in enteric or other nerve cells and that a viral protein that is produced in infected cells but which is not incorporated into viral particles (ORF61p) is required for VZV to give rise to an infection that produces more virus and kills infected cells (lytic infection) or to reactivate from latency in nerve cells.

描述（由申请人提供）：肠神经系统（ENS）可以控制肠道行为，而无需大脑或脊髓的输入。功能性的ENS对于生命至关重要，当异常时会引起不适，并可能导致胃肠道运动，分泌和炎症的病理生理学或严重程度。我们最近发现，在大多数经历过自然水痘或接受水疗疫苗的个体中，水痘带状疱疹病毒（VZV）在人类肠神经元内建立潜伏期。此外，VZV已与免疫功能低下个体中致死性假期结构的发生有关。先前曾探索过VZV访问ENS的途径，也没有在肠神经元中（“肠goster”）重新激活的频率或后果。当前的提案旨在测试内脏传入神经中转运的假设向ENS进行VZV，无细胞病毒体在肠神经元中产生潜伏期，并且必须在神经元中表达非结构性VZV ORF61蛋白，以使VZV启用VZV以表现出裂纹感染或从延伸延伸。尽管VZV表现出对人类细胞的明显偏好，但我们开发了允许在体外和体内研究的ENS的VZV感染的动物模型。根据条件，VZV概括了从豚鼠或小鼠分离的肠神经元中的潜在，裂解和重新激活的感染，当引入肠中时，VZV在豚鼠ensu中建立潜伏期。该提案具有3个具体目标：（1）VZV可以在感觉神经纤维中从皮肤传播到ENS吗？初步研究已经确定了在皮肤和肠道上都投射的背途径神经节中的神经元。我们将确定当将VZV引入皮肤时是否在ENS中建立潜在感染，以及VZV感染的神经末端是否释放了无感染性细胞的VZV，从而跨越突触间隙以将潜在感染传递到靶神经元。 （2）病毒血症可以建立肠神经元的潜在VZV感染吗？我们将确定VZV感染的T淋巴细胞是释放无感染性细胞的VZV，以及它们是否可以直接在肠神经元中或通过粘膜上皮细胞或皮肤的感染而间接地在肠神经元中建立潜伏期。初步研究表明，静脉注射VZV感染的外周血单核细胞后豚鼠肠神经元中存在VZV DNA。 （3）VZV ORF61蛋白表达对于肠神经元中裂解感染的表现是否需要？我们将研究缺乏ORF61建立肠神经元裂解感染或从潜伏期重新激活的VZV突变体的能力。了解ENS的VZV感染的重要性增强了肠神经元中VZV的重新激活的可能性，可能有助于诸如肠胃疾病，炎症综合征，炎症性肠病，特分病毒性胃癌和慢性遗传性遗传性遗传性的疾病疾病的发病机理。 公共卫生相关性：肠道包含一个大型且复杂的神经细胞网络，称为肠神经系统（ENS），该网络能够控制肠道行为，而无需大脑或脊髓的输入。 ENS促成了许多干扰ENS功能的疾病的基础，例如肠易激综合征，炎症性肠病，特发性胃轻瘫和慢性肠道假性结构尚不清楚。我们最近发现，水痘带状疱疹病毒（VZV）在人ENS的神经细胞内建立潜伏期。以前都没有探索过这种现象的原因和后果，因为肠道神经细胞中的潜在VZV尚未发生。当前的建议旨在确定VZV如何获得ENS的访问权限，我们将利用肠神经细胞来测试仅VZV的无细胞颗粒形式能够在肠或其他神经细胞中建立潜伏期的假设，并且在感染中产生的病毒蛋白不合时间就产生了aNE的病毒蛋白，从而导致了Infire and Fire and Fire and Fire（ORF61P）IS（ORF61p）IS INS INS INS INS INS INS INS INS INS INS INS INS INS INS INS INS INS-VZ VZ VZ。感染的细胞（裂解感染）或因神经细胞潜伏期而重新激活。