Radhakrishnan Sujatha, Otte Jessica, Enam Sahnila, Del Valle Luis, Khalili Kamel, Gordon Jennifer
Center for Neurovirology and Cancer Biology, College of Science and Technology, Temple University, Philadelphia, Pennsylvania 19122, USA.
J Virol. 2003 Oct;77(19):10638-44. doi: 10.1128/jvi.77.19.10638-10644.2003.
Cell-type-specific transcription of the JC virus (JCV) promoter in glial cells initiates a series of events leading to viral replication in the brain and the development of the fatal demyelinating disease progressive multifocal leukoencephalopathy (PML) in patients with neurologic complications due to infection with human immunodeficiency virus type 1. Here we employed an in vitro infection of primary cultures of human astrocytes to compare the transcriptional profile of cellular genes after JCV infection by using an oligonucleotide-based microarray of 12600 genes. Transcription of nearly 355 genes was enhanced and expression of 130 genes was decreased to various degrees. Many transcripts that were increased upon JCV infection were found to encode proteins with properties that suggest their involvement in cell proliferation, including cyclin A and cyclin B1; signaling pathways, such as transforming growth factor beta receptor 1, platelet-derived growth factor receptor and fibroblast growth factor family receptor; and other regulatory events, such as inflammatory responses, including cyclo-oxygenase-2 (Cox-2). Microarray-based data for several cell cycle-regulatory genes were further examined by using Western blot analysis of in vitro infected astrocytes harvested early and late during the infection. Results demonstrate that protein levels of all upregulated genes were found to increase at some point during the infection time course. In parallel, immunohistochemical assessment of cell cycle proteins, including cyclins A, B1, E, and Cdk2, showed positive staining of astrocytes within PML lesions of brain tissue from patients with neuro-AIDS. Microarray analysis was found to be a useful predictor of gene expression in infected cells; however, it may not directly correlate with protein levels during infection with JCV.
在神经胶质细胞中，JC病毒（JCV）启动子的细胞类型特异性转录引发了一系列事件，导致病毒在大脑中复制，并在因感染1型人类免疫缺陷病毒而出现神经系统并发症的患者中引发致命的脱髓鞘疾病——进行性多灶性白质脑病（PML）。在此，我们利用人星形胶质细胞原代培养物进行体外感染，通过使用包含12600个基因的基于寡核苷酸的微阵列，比较JCV感染后细胞基因的转录谱。近355个基因的转录增强，130个基因的表达不同程度降低。JCV感染后增加的许多转录本编码的蛋白质具有提示其参与细胞增殖的特性，包括细胞周期蛋白A和细胞周期蛋白B1；信号通路，如转化生长因子β受体1、血小板衍生生长因子受体和成纤维细胞生长因子家族受体；以及其他调节事件，如炎症反应，包括环氧化酶-2（Cox-2）。通过对感染早期和晚期收获的体外感染星形胶质细胞进行蛋白质印迹分析，进一步检测了几个细胞周期调节基因基于微阵列的数据。结果表明，在感染时间进程的某个时间点，所有上调基因的蛋白质水平均升高。同时，对包括细胞周期蛋白A、B1、E和细胞周期蛋白依赖性激酶2在内的细胞周期蛋白进行免疫组织化学评估，显示神经艾滋病患者脑组织PML病变内的星形胶质细胞呈阳性染色。发现微阵列分析是感染细胞中基因表达的有用预测指标；然而，它可能与JCV感染期间的蛋白质水平没有直接相关性。
