The cell biology of Theiler's virus persistence in CNS

泰勒病毒在中枢神经系统中持续存在的细胞生物学

• 批准号: 7244401
• 负责人: Karla Kirkegaard
• 金额: $ 33.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7244401
• 关键词: 5-(6)-carboxyfluorescein diacetate succinimidyl ester; Address; Animal Model; Appearance; Astrocytes; Blood; Bone Marrow; Brain; Cells; Cellular biology; Chimera organism; Chronic; Coculture Techniques; Defect; Demyelinations; Disease; Ensure; Face; Family Picornaviridae; Female; Fluorescent Dyes; Frequencies; Gray unit of radiation dose; Heterozygote; Hypersensitivity; Immune; Immune response; Immune system; In Vitro; Inbred C3H Mice; Individual; Infection; Inflammation; Inflammatory; Label; Lesion; Link; Mediating; Methods; Modeling; Monitor; Mus; Mutant Strains Mice; Mutation; Myelin; Myelin Sheath; Neuroglia; Neurons; Oligodendroglia; Peripheral; Population; Predisposition; Rate; Recruitment Activity; Resistance; Role; Spinal Cord; TMEV; Testing; Time; Tissues; Viral; Viral Genome; Virus; Virus Diseases; Week; Wild Type Mouse; Work; X Inactivation; base; cell type; in vivo; macrophage; migration; monocyte; myelination; permissiveness; pressure; research study; spinal cord white matter; trafficking; white matter

DESCRIPTION (provided by applicant): Theiler's virus, a picornavirus, causes a persistent infection of the spinal cord of genetically susceptible mice that is accompanied by chronic infection and primary demyelination. As such, it provides one of the best animal models for immune-mediated demyelination. Theiler's virus first infects neurons, but later persists in oligodendrocytes and macrophage/macroglial cells. This shift from gray to white matter coincides with the appearance of virus-specific immune responses, suggesting that immune pressure keeps the virus out of neurons but is unable to clear it from glial cells. In this application, we propose to test a model in which the virus traffics sequentially from neurons to oligodendrocytes to macrophages, with oligodendrocytes providing an obligatory intermediate in the face of immune pressure. First, we will monitor the time course of Theiler's infection of various cell types in wild-type C3H and in shiverer and rumpshaker mice, which have different severe myelin defects and are resistant to persistent infection. We will examine whether oligodendrocytes, macrophages or both are never infected, or infected only transiently, in the resistant mutant mice. Bone marrow transfer experiments will reveal whether CNS- or bone marrow-derived cells from shiverer and rumpshaker mice confer resistance to persistent infection and in vitro culture of oligodendrocytes and macrophages will reveal whether the intrinsic permissiveness of these cells to viral infection is altered by the mutations. Second, we will determine whether myelin is the portal of entry of the virus into the white matter. We will express viral genomes specifically in neurons in mixed cultures of oligodendrocytes and neurons, and track viral spread into oligodendrocytes in the presence and absence of successful myelination. Experiments performed with female rumpshaker heterozygotes will afford us the possibility to work with natural chimeras in vivo, because the rumpshaker mutation is X-linked. Finally, we will use in vivo labeling methods to monitor the migration of blood-borne macrophages into inflammatory CNS lesions, and determine the frequency with which these newly arrived cells become infected. This will test the possibility that persistence is achieved, in part, by the chronic inflammation that ensures the existence of a renewable host population. Together, this model may prove a paradigm for the functional transfer of infectious material from neurons to oligodendrocytes, and from thence to cells of the immune system.

描述（由申请方提供）：泰勒病毒是一种小核糖核酸病毒，可引起遗传易感小鼠脊髓的持续感染，并伴有慢性感染和原发性脱髓鞘。因此，它为免疫介导的脱髓鞘提供了最好的动物模型之一。Theiler病毒首先感染神经元，但随后在少突胶质细胞和巨噬细胞/大胶质细胞中持续存在。这种从灰色到白色物质的转变与病毒特异性免疫反应的出现相吻合，这表明免疫压力使病毒远离神经元，但无法将其从神经胶质细胞中清除。在这个应用中，我们建议测试一个模型，其中病毒依次从神经元到少突胶质细胞再到巨噬细胞，少突胶质细胞在面对免疫压力时提供了一个强制性的中间体。首先，我们将监测野生型C3 H和颤抖和摇摆小鼠中各种细胞类型的Theiler感染的时间过程，这些小鼠具有不同的严重髓鞘缺陷并且对持续感染具有抵抗力。我们将研究是否少突胶质细胞，巨噬细胞或两者都从未感染，或感染只是短暂的，在耐药突变小鼠。骨髓转移实验将揭示是否CNS或骨髓来源的细胞从shiverer和rumpshaker小鼠赋予持久性感染的抵抗力和少突胶质细胞和巨噬细胞的体外培养将揭示是否这些细胞的内在许可病毒感染的突变被改变。其次，我们将确定髓磷脂是否是病毒进入白色物质的入口。我们将在少突胶质细胞和神经元的混合培养物中的神经元中特异性地表达病毒基因组，并在存在和不存在成功髓鞘形成的情况下跟踪病毒扩散到少突胶质细胞中。用雌性rumpshaker杂合子进行的实验将为我们提供在体内研究天然嵌合体的可能性，因为rumpshaker突变是X连锁的。最后，我们将使用体内标记方法来监测血液传播的巨噬细胞迁移到炎症性CNS病变，并确定这些新到达的细胞被感染的频率。这将测试持久性的可能性，部分是通过确保可再生宿主种群存在的慢性炎症来实现的。总之，这个模型可以证明一个范例的功能转移的感染性物质从神经元到少突胶质细胞，并从那里到免疫系统的细胞。