MOLECULAR PATHOGENESIS OF JC VIRUS & PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY

JC 病毒的分子发病机制

• 批准号: 3760211
• 负责人: E O MAJOR
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3760211
• 关键词: DNA footprinting; Polyomavirus hominis 2; analytical method; biopsy; gene expression; genetic regulatory element; genetic strain; host organism interaction; human tissue; molecular pathology; neurotropic virus; nucleic acid sequence; polymerase chain reaction; progressive multifocal leukoencephalopathy; reporter genes; transcription factor; virus DNA; virus genetics

The presence of JCV in peripheral blood lymphocytes (PBLs) in both viral induced progressive multifocal leukoencephalopathy (PML) and non-PML patients has strengthened the concept that JCV is carried to the brain by a hematogenous route. At the same time, it has focused a part of our studies to develop a quantitative analysis of the level of JCV in these cells and other tissues. It may be important in helping to assess the risk of non-PML patients and to evaluate the affect nucleotide analogs for the treatment of PML. Noncompetitive and competitive or quantitative PCR (QPCR) analysis procedures are being developed to ascertain the level of JCV present in different tissues. Control JCV DNA templates are being constructed to serve as external or internal standards for the quantitation of the virus. These procedures utilize the same primer set used to detect JCV in patient samples by PCR analysis which eliminates the possible differential hybridization kinetics when using different primer sets. A rapid and sensitive chemiluminescent procedure is being used to analyze the PCR products in these studies. An examination of the nucleotide sequences present in the prototype (Mad1) and brain-type (Mad 8B) JCV is being made to assess the role of these sequences in the pathogenicity of JCV. One major difference is the presence of a 23bp sequence which is present in most strains of JCV isolated from brain biopsies from PML patients. This 23bp sequence has a putative binding site for the transcription factor SP1. However, we have found that SP1 binds to this sequence very weakly as seen by competitive binding and DNase 1 protection assays. In addition, construction of CAT reporter plasmids containing either the Mad1 or Mad8B regulatory region has shown that the Mad1 sequence is much more active that from Mad8B. Further studies are underway to understand the role of the 23-bp sequence in the expression of the Mad8B strain in different tissues.

两种病毒感染者的外周血淋巴细胞（PBL）中JCV的存在均为阴性。 诱导的进行性多灶性白质脑病（PML）和非PML 患者加强了JCV被携带到大脑的概念， 通过血行途径 与此同时，它集中了我们的一部分， 研究开发定量分析的水平JCV在这些 细胞和其他组织。 这可能有助于评估 非PML患者的风险，并评估受影响的核苷酸类似物 用于治疗PML。非竞争性和竞争性或定量 正在制定PCR（QPCR）分析程序，以确定 JCV存在于不同的组织中。 对照JCV DNA模板正在 作为外部或内部标准， 病毒的定量。 这些程序使用相同的引物组 用于通过PCR分析检测患者样本中的JCV， 当使用不同的杂交剂时， 引物组。一种快速灵敏的荧光检测方法 用于分析这些研究中的PCR产物。 的检查 存在于原型（Mad 1）和脑型（Mad）中的核苷酸序列 8B）JCV正在评估这些序列在 JCV的致病性 一个主要的区别是存在一个23 bp的 该序列存在于从脑中分离的大多数JCV菌株中 PML患者的活检。 该23 bp的序列具有假定的结合 转录因子SP1的位点。 但是，我们发现SP1 通过竞争性结合可以看出，与该序列的结合非常弱， DNA酶1保护测定。此外，CAT报告器的构建 含有Mad 1或Mad 8B调节区的质粒已经显示 Mad 1序列比Mad 8B序列更活跃。 进一步 研究正在进行中，以了解23-bp序列在 Mad 8B菌株在不同组织中的表达。