MOLECULAR PATHOGENESIS OF JC VIRUS & PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY

两种病毒外周血淋巴细胞中JCV的存在 诱发进行性多灶性白质脑病与非进行性多灶性白质脑病 患者强化了JCV被带到大脑的概念 通过一种血源性途径。同时，它也集中了我们的一部分 对这些人的JCV水平进行定量分析的研究 细胞和其他组织。它可能在帮助评估 非PML患者的风险及其影响核苷酸类似物的评估 用于治疗PML。非竞争性的、竞争性的或数量的 多聚酶链式反应(QPCR)分析程序正在开发中，以确定该水平 JCV在不同组织中均有表达。控制JCV DNA模板正在进行中 被构造为用作 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，消除了 不同品种间差异杂交的可能动力学 底漆套装。一种快速而灵敏的化学发光程序正在进行中 用于分析这些研究中的PCR产物。一项关于 存在于原型(Mad1)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23个基点 从脑中分离的JCV存在于大多数毒株中的序列 PML患者的活组织检查。这个23bp的序列有一个假定的结合 转录因子SP1的位置。然而，我们发现SP1 与这个序列的结合非常弱，通过竞争结合和 DNase1保护性检测。此外，CAT记者的建设 含有Mad1或Mad8B调节区的质粒已显示 Mad1序列比Mad8B序列更活跃。进一步 目前正在进行研究，以了解23-bp序列在生物多样性中的作用。 Mad8B株在不同组织中的表达。

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.