Development of a Mouse Model of Progressive Multifocal Leukoencephalopathy

进行性多灶性白质脑病小鼠模型的建立

基本信息

批准号：
8649144
负责人：
Elizabeth Leigh Frost
金额：
$ 2.83万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8649144
关键词：
AIDS/HIV problem Acute Affect Affinity Allografting Amino Acid Substitution Animal Model Antibodies Antiviral Agents Autoimmune Diseases Autoimmune Process BCL6 gene BK Virus Binding Binding Sites Bone Marrow Brain CD4 Positive T Lymphocytes CD8B1 gene Capsid Capsid Proteins Cells Central Nervous System Diseases Central Nervous System Infections Cerebrospinal Fluid Cytolysis DNA Sequence Rearrangement Demyelinations Depressed mood Development Disabled Persons Disease Disease Progression Evaluation Flow Cytometry Functional RNA Functional disorder Goals Hematologic Neoplasms Hematological Disease Histopathology Human Immune Immune system Immunocompetent Immunofluorescence Microscopy Immunologic Monitoring Immunosuppression Immunosuppressive Agents Incidence Individual Infection Infection Control Inflammatory Integrin alpha4beta1 JC Virus Kidney Kidney Diseases Laboratories Lesion Leucocytic infiltrate Life Lytic Maintenance Maps Mediating Memory Modeling Monoclonal Antibodies Monoclonal Antibody Therapy Multiple Sclerosis Mus Mutation Neuraxis Neurodegenerative Disorders Neuroglia Neurologic Oligodendroglia Organ Pathogenesis Patients Pharmaceutical Preparations Play Point Mutation Polymerase Chain Reaction Polyomavirus Polyomavirus Infections Population Positioning Attribute Predisposition Progressive Multifocal Leukoencephalopathy Relapsing-Remitting Multiple Sclerosis Risk Risk Factors Role Satellite Viruses Severities Sialic Acids Site Species Specificity T cell response T memory cell T-Lymphocyte Testing Tissues Transplantation Tropism Urinary tract Urine Viral Viral Load result Virulence Virus Virus-like particle Work adverse outcome base cell type central nervous system demyelinating disorder central nervous system injury handicapping condition humanized monoclonal antibodies immune function immunosuppressed kidney allograft leukemia migration mortality mouse model mouse polyomavirus mutant natalizumab neuropathology neurotropic neurovirulence pathogen prevent public health relevance receptor binding trafficking treatment duration

项目摘要

DESCRIPTION (provided by applicant): Human JC polyomavirus (JCV) is the causative agent of progressive multifocal leukoencephalopathy (PML), a life-threatening demyelinating disease of the central nervous system (CNS) resulting from infection of oligodendrocytes. JCV infection is prevalent in the human population, where it persists lifelong as an asymptomatic infection in healthy individuals. JCV-induced PML manifests in individuals immunosuppressed by HIV/AIDS, leukemia, allograft transplantation, or, more recently, monoclonal antibody-mediated therapy for autoimmune and inflammatory diseases. Natalizumab, a monoclonal antibody therapy for treatment of multiple sclerosis, carries a risk for PML; incidence of PML in these patients rises with duration of treatment. This therapy is intended to prevent migration of autoimmune T cells to their target tissues, but blockade of trafficking of JCV-specific T cells into the CNS is generally considered its adverse consequence. JCV isolated from PML patients were found to have mutations in the major capsid protein VP1 and the non-coding control region (NCCR), implicating virus-associated risk factors in addition to host immune compromise. Several specific VP1 mutations have been identified, all of which are single amino acid substitutions that map to sialic acid binding sites, which are involved in attachment and entry of virus into host cells. Species-specificity of polyomaviruses has severely impeded development of a tractable JCV-PML animal model, and as a result our understanding of the pathogenesis of PML is limited. For example, it remains to be determined if mutations in VP1 alter virulence, and the role that immunosuppression plays in disease progression. The goal of this project is to develop a model of PML in mice using mouse polyomavirus (MPyV) to investigate how virus capsid mutations, adaptive immune system deficiencies, and treatment with natalizumab contribute to induction of polyomavirus- induced CNS disease. Introduction of a PML-associated substitution into an analogous site in VP1 of MPyV is hypothesized to increase the severity of CNS disease in mice by making MPyV neurotropic, allowing infection and damage of glial cells, including oligodendrocytes. Shifts in tropism will be determined by quantitative polymerase chain reaction (PCR) to assess viral replication efficiency, immunofluorescence microscopy to define infected CNS cells, and histopathologic evaluation of CNS injury. As in human PML, CNS infection is likely exacerbated by immunosuppression. Using therapy similar to natalizumab in an MPyV CNS infection model, flow cytometry-based phenotypic and functional analyses will be used to determine the mechanisms by which immunosuppression contributes to pathogenesis. Successful development of this MPyV infection model would provide the opportunity to explore mechanisms of polyomavirus CNS pathogenesis in a natural host.

描述（由申请人提供）：人类JC多瘤病毒（JCV）是进行性多焦点白细胞术（PML）的病因，这是一种威胁生命的中枢神经系统（CNS）的脱髓鞘疾病，该疾病是由少突endrocytes感染引起的。 JCV感染在人群中普遍存在，在健康个体中，它一直是无症状的感染。 JCV诱导的PML在因HIV/AIDS，白血病，同种异体移植移植或最近的单克隆抗体介导的自身免疫性和炎症性疾病的疗法的免疫抑制的个体中表现出来。 Natalizumab是一种用于治疗多发性硬化症的单克隆抗体疗法，它具有PML的风险。这些患者中PML的发病率随着治疗时间而增加。该疗法旨在防止自身免疫性T细胞迁移到其靶组织，但是将JCV特异性T细胞运输到中枢神经系统中的封锁通常被认为是其不良后果。发现从PML患者中分离出的JCV在主要的衣壳蛋白VP1和非编码控制区域（NCCR）中具有突变，除了宿主免疫妥协外，还暗示了与病毒相关的危险因素。已经鉴定出了几种特定的VP1突变，所有这些突变都是映射到唾液酸结合位点的单个氨基酸取代，这些氨基酸取代与病毒的附着和进入宿主细胞有关。多瘤病毒的物种特异性严重阻碍了可拖动的JCV-PML动物模型的发展，因此，我们对PML发病机理的理解是有限的。例如，VP1中的突变是否改变了毒力，以及免疫抑制在疾病进展中起的作用是否尚待确定。该项目的目的是使用小鼠多瘤病毒（MPYV）在小鼠中开发PML模型，以研究病毒衣壳突变，适应性免疫系统缺乏症以及用纳他蛋白单抗治疗如何有助于诱导多瘤病毒诱导的CNS疾病。假设将与PML相关的取代引入MPYV VP1中的类似位点可以通过使MPYV神经性化，从而增加小鼠中枢神经系统疾病的严重程度，从而允许感染和神经胶质细胞的损害，包括少突胶质细胞。向流主义的转移将由定量聚合酶链反应（PCR）确定，以评估病毒复制效率，免疫荧光显微镜以定义感染的CNS细胞以及CNS损伤的组织病理学评估。与人类PML一样，CNS感染可能会因免疫抑制而加剧。在MPYV CNS感染模型中，使用类似于Natalizumab的治疗，将使用基于流式细胞术的表型和功能分析来确定免疫抑制有助于发病机理的机制。该MPYV感染模型的成功开发将为探索自然宿主中多瘤病毒CNS发病机理的机制提供机会。