CHRONIC VIRAL INFECTIONS--MOLECULAR BIOLOGY OF HUMAN JC VIRUS

我们继续对JC病毒和人脑胶质细胞的分子病理学进行研究 培养中的细胞，以及最近从人脑组织切片中 进行性多灶性白质脑病(PML)感染患者。 实验有细胞内水平的重点描述的性质 病毒在中枢神经系统的持续存在，PML作为脱髓鞘的发病机制 疾病，以及对病毒和细胞的通用调节机制 基因参与其中。我们的成果包括解决了最大的问题 与JC合作已提出，即宿主和组织限制 原代培养人胎儿神经胶质细胞的生长。我们已经建立了 利用突变体获得永生化的人胎儿星形胶质细胞系 SV40的转化基因。这些细胞能够产生具有传染性的JCV。 并使我们能够研究CV DNA复制和T的动力学 蛋白质合成。我们还发现，SV40T蛋白在 人神经胶质细胞与神经胶质细胞蛋白P53的复合体 细胞中的这种蛋白质。JCVT蛋白似乎不是复合体 与P53蛋白的关系，可能是由于确认了 SV40和JCV的T蛋白。这种差异的更多证据出现了 我们的实验表明JCV的环丁残基发生了烷基化反应 T蛋白是分离免疫沉淀T细胞所必需的。 蛋白质变性凝胶电泳法。这并不是必须的 或者是SV40的T蛋白，或者是相关的人类BK病毒。的研究 来自PML患者的石蜡包埋或冰冻脑组织显示 用原位杂交技术检测JCV DNA的存在及其与 用免疫细胞化学试验检测病毒衣壳抗原。这些 研究还表明，对偶树突胶质细胞是 JCV基因表达，但PML斑块中存在奇怪的星形细胞 显示JCV DNA和衣壳抗原。后者发现问题的作用是 JC病毒在恶性胶质瘤的发病机制中可能有一定作用 其他人提出的总体人口。

Our studies continue on the molecular pathology of JC virus and human glial cells in culture, and more recently, with human brain tissue sections from infected patients wth progressive multifocal leukoencephalopathy (PML). Experiments have focues at the intracellular level describing the nature of viral persistance in the CNS, pathogenesis of PML as a demyelinating disease, and the mechanisms of generic regulation of the viral and cellular genes involved. Our results include solving the biggest problems that working with JC has presented, namely the host and tissue restriction for growth to primary human fetal glial cells in culture. We have estalbished an immortalized line of human fetal astroglial cells using a mutant transforming gene of SV40. These cells are able to produce infectious JCV and have allowed us to examine the kinetics of CV DNA replication and T protein synthesis. We have also found that the SV40 T protein made in human glial cells complexes with a glial cell protein, p53, and stablizes this protein in the cell. The JCV T protein does not appear to complex with the p53 protein, perhaps due to confirmational differences between the T proteins of SV40 and JCV. Additional evidence for such differences came from our experiments suggesting alkylation of cycteine residues of the JCV T protein was necessary in order to resolve the immunoprecipitated T protein using denaturing gel electrophoresis. This was not necessary for either the T proteins of SV40 or the related human BK virus. Studies of paraffin embedded or fozen brain tissue from PML patients showed the presence of JCV DNA using in situ hyridization and correlated with the detection of viral capsid antigen using immunocytochemical tests. These studies also revealed that aligodendroglian cells are the main target for JCV gene expression but that bizarre astrocyctes present in PML plaques showed JCV DNA and capsid antigen. This latter finding questions role the of JC virus may have in the pathogenesis of malignant gliomas in the general population that has been suggested by others.