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A Mouse Model to Define Immunovirologic Determinants of Polyomavirus CNS Disease

定义多瘤病毒中枢神经系统疾病免疫病毒学决定因素的小鼠模型

基本信息

批准号：
8853962
负责人：
Aron Eliot Lukacher
金额：
$ 33.47万
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：
2014-06-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8853962
关键词：
A Mouse AIDS/HIV problem Acute Address Adhesions Amino Acid Substitution Animal Model Antibodies Antibody Response Antiviral Agents Autoimmune Process BK Virus Binding Blood Blood - brain barrier anatomy Bone Marrow Brain Brain Diseases CD4 Positive T Lymphocytes CD8B1 gene Capsid Capsid Proteins Cells Central Nervous System Diseases Complex Complication Cutaneous Demyelinating Diseases Demyelinating Encephalopathy Depressed mood Disease Elements Environment Extravasation Family Flow Cytometry Functional disorder Genome Genomic DNA Genotype Goals Health Hematologic Neoplasms Heterogeneity High Prevalence Human Humoral Immunities Hybrids Immune Immunologic Monitoring Immunologics Immunosuppression Immunosuppressive Agents Individual Infection Inflammatory Integrin alpha4beta1 Integrins JC Virus Kidney Diseases Libraries Life Lung MHC Class I Genes Maintenance Mediating Memory Modeling Monitor Monoclonal Antibodies Multiple Sclerosis Mus Mutagenesis Mutation Neuroglia Neurotropism Nonstructural Protein Organ Pathogenesis Patients Play Polyomavirus Polyomavirus Infections Polysaccharides Progressive Multifocal Leukoencephalopathy Receptor Cell Recording of previous events Regimen Relapse Relapsing-Remitting Multiple Sclerosis Replication Origin Risk Risk Factors Role Sialic Acids Skin Tissue Specificity T memory cell T-Lymphocyte Testing Therapeutic immunosuppression Tissues Transgenic Mice Transplant Recipients Tropism Tysabri Urinary tract Variant Viral Viral Tumor Antigens Viral load measurement Virulence Virus base brain tissue cell motility central nervous system injury exhaustion humanized monoclonal antibodies immune function immunosuppressed in vivo kidney allograft mouse model mouse polyomavirus mutant natalizumab neurotropic neurovirulence neutralizing antibody next generation sequencing novel pathogen pre-clinical prevent residence seropositive white matter

项目摘要

DESCRIPTION (provided by applicant): Progressive Multifocal Leukoencephalopathy (PML), a demyelinating encephalopathy caused by the human JC polyomavirus, is a life-threatening complication of natalizumab therapy for relapsing multiple sclerosis. Seropositivity for JCV, prior immunosuppression, and > 24 month of natalizumab treatment are the only identified factors known to increase risk for PML, but their utility is limited because of the high prevalence of JCV infection and the heterogeneity of immunosuppressive regimens patients receive before starting natalizumab. No anti-JCV agents are available. Elucidation of the pathogenesis of PML and immunosurveillance mechanisms that keep JCV replication in check are required to adequately assess risk for PML in MS individuals receiving natalizumab. Polyomaviruses are highly species specific, with humans being the only host reservoir for JCV. A tractable animal model is urgently needed to understand the immunologic and virologic determinants of JCV-induced PML, and to provide a preclinical platform to assess the in vivo efficacy of novel anti-JCV compounds. Using mouse polyomavirus (MPyV), we have developed a mouse model of polyomavirus-induced CNS disease. We propose applying this MPyV-CNS infection model to address two Aims based on the following hypotheses: (1) by blocking T cell extravasation across the blood-brain barrier into the CNS parenchyma, natalizumab prevents maintenance of brain-resident virus-specific CD8 T cells required for antiviral immunosurveillance; and (2) because JCV in PML patients harbor novel mutations in their capsid protein, the dominant target for humoral-mediated immune defense, virus-neutralizing antibodies select viral variants that acquire neurotropism. For Aim 1, we have constructed MHC class I and class II tetramers to visualize MPyV-specific CD8 and CD4 T cells by flow cytometry, and have developed an MPyV- specific TCR transgenic mouse to monitor the fate of anti-MPyV CD8 T cells in the CNS. For Aim 2, we will create a library of capsid mutant MPyVs by PCR-based random mutagenesis, and by combining iterative virus passaging in vivo with next-generation sequencing, determine whether neutralizing MPyV antibodies select mutations that confer neurovirulence. We will also attempt to create a hybrid virus expressing the capsid proteins of JCV (to confer JCV host cell specificity) and having the noncoding control elements, origin of replication, and nonstructural proteins of MPyV (to enable replication in the mouse). This novel hybrid virus will allow us to determine the functional significance of the capsid mutations in JCV-PML variant viruses.

描述(申请人提供)：进行性多灶性白质脑病(PML)是一种由人类JC多瘤病毒引起的脱髓鞘脑病，是复发性多发性硬化症那他珠单抗治疗的一种危及生命的并发症。JCV的血清阳性、既往的免疫抑制和接受Natalizumab治疗24个月是唯一已知的增加PML风险的因素，但由于JCV感染的高流行率和患者在开始接受Natalizumab之前接受的免疫抑制方案的异质性，它们的作用有限。没有可用的抗JCV试剂。需要阐明PML的发病机制和控制JCV复制的免疫监测机制，以充分评估接受Natalizumab治疗的MS患者PML的风险。多瘤病毒具有高度的物种特异性，人类是JCV的唯一宿主宿主。迫切需要一个易于处理的动物模型来了解JCV诱导的PML的免疫学和病毒学决定因素，并提供一个临床前平台来评估新型抗JCV化合物的体内疗效。利用小鼠多瘤病毒(MPyV)建立了多瘤病毒诱导的中枢神经系统疾病的小鼠模型。我们建议应用这种MPyV-CNS感染模型来解决基于以下假设的两个目标：(1)通过阻止T细胞跨越血脑屏障渗入中枢神经系统实质，Natalizumab阻止抗病毒免疫监视所需的脑驻留病毒特异性CD8 T细胞的维持；以及(2)由于PML患者的JCV在其衣壳蛋白中含有新的突变，衣壳蛋白是体液介导免疫防御的主要靶点，病毒中和抗体选择获得神经趋向性的病毒变体。对于目的1，我们构建了MHC I类和II类四聚体，通过流式细胞术显示MPyV特异性的CD8和CD4T细胞，并建立了MPyV特异性TCR转基因小鼠来监测抗MPyV CD8T细胞在中枢神经系统中的命运。对于目标2，我们将通过基于聚合酶链式反应的随机突变建立衣壳突变MPyV文库，并通过体内迭代病毒传代和下一代测序相结合，确定中和MPyV抗体是否选择赋予神经毒力的突变。我们还将尝试创建一种表达JCV衣壳蛋白的杂交病毒(以赋予JCV宿主细胞特异性)，并具有MPyV的非编码控制元件、复制起点和非结构蛋白(以在小鼠中复制)。这种新型的杂交病毒将使我们能够确定JCV-PML变异病毒衣壳突变的功能意义。