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：
9244865
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/9244865
关键词：
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 CD8-Positive T-Lymphocytes CD8B1 gene Capsid Capsid Proteins Cells Central Nervous System Diseases Central Nervous System Infections Complex Complication Cutaneous Demyelinating Diseases Demyelinating Encephalopathy Depressed mood Disease Elements Environment Extravasation Family Flow Cytometry Functional disorder Genome Genomic DNA Genotype Goals Hematologic Neoplasms Heterogeneity High Prevalence Human Humoral Immunities Hybrids Immune Immunologic Monitoring Immunologics Immunosuppression Immunosuppressive Agents Impairment Individual Infection Inflammatory Integrin alpha4 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 Mutant Strains Mice 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 immunological status immunoregulation immunosuppressed in vivo kidney allograft mouse model mouse polyomavirus multiple sclerosis patient mutant natalizumab neurotropic neurovirulence neutralizing antibody next generation sequencing novel pathogen pre-clinical prevent public health relevance residence seropositive therapy development virology 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（由申请人提供）：渐进性多灶性白血病（PML）是由人类JC多瘤病毒引起的一种脱髓性脑病，是Natalizumab疗法的生命并发症，用于复发多发性硬化症。 Natalizumab治疗的JCV，先前的免疫抑制和> 24个月> 24个月的血清阳性是已知的唯一确定的因素，这会增加PML的风险，但由于JCV感染的高流行和免疫抑制治疗方案患者的异质性，其效用受到限制。没有抗JCV代理。需要检查PML和免疫监视机制的发病机制，以使JCV复制进行检查以充分评估接受Natalizumab的MS个体的PML风险。多瘤病毒是高度物种特异性的，人类是JCV的唯一宿主储层。迫切需要一种可拖动的动物模型，以了解JCV诱导的PML的免疫和病毒学决定因素，并提供一个临床前平台来评估新型抗JCV化合物的体内功效。使用小鼠多瘤病毒（MPYV），我们开发了一种多瘤病毒诱导的CNS疾病的小鼠模型。我们建议将此MPYV-CNS感染模型应用于以下假设来解决两个目的：（1）通过将跨血脑屏障的T细胞渗入到CNS实质中，Natalizumab可阻止维持抗病毒免疫免疫性抗病性免疫抗性所需的脑部耐脑脑遗留病毒CD8 T细胞；（2）由于PML患者的JCV在其衣壳蛋白中具有新型突变，因此是体液介导的免疫防御的主要靶标，因此病毒中和抗体选择获得神经旋转的病毒变体。对于AIM 1，我们构建了MHC I类和II类四聚体，通过流式细胞仪可视化MPYV特异性CD8和CD4 T细胞，并开发了一种MPYV-特异性TCR转基因小鼠，以监测CNS中抗MPYV CD8 T细胞的命运。对于AIM 2，我们将通过基于PCR的随机诱变来创建一个capsid突变型MPYV的库，并通过将体内的迭代病毒与下一代测序结合起来，确定中和MPYV抗体是否选择赋予神经毒害的突变。我们还将尝试创建一种表达JCV的衣壳蛋白（赋予JCV宿主细胞特异性）的混合病毒，并具有非编码控制元件，复制的起源和MPYV的非结构蛋白（在小鼠中启用复制）。这种新型的杂种病毒将使我们能够确定JCV-PML变体病毒中衣壳突变的功能意义。