Proteins Mediating Interaction of HIV-1 and JCV in CNS

介导中枢神经系统中 HIV-1 和 JCV 相互作用的蛋白质

• 批准号: 7382577
• 负责人: EDWARD M. JOHNSON
• 金额: $ 42.35万
• 依托单位: EASTERN VIRGINIA MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-02-01 至 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7382577
• 关键词: Acquired Immunodeficiency Syndrome; Affect; Automobile Driving; Binding; Biological Assay; Brain; Categories; Cells; Conditioned Culture Media; DNA biosynthesis; Data; Development; Environment; Exposure to; Gene Chips; Genes; Genetic Transcription; Goals; HIV; HIV tat Protein; HIV-1; Immunocompromised Host; Immunohistochemistry; Immunologic Deficiency Syndromes; Immunomodulators; Immunosuppression; Incidence; Individual; Infection; Intervention; JC Virus; Knock-out; LacZ Genes; Late Gene Transcriptions; Lesion; Leukoencephalopathy; Localized; Mediating; Mediation; Methods; Microglia; Mus; Neurodegenerative Disorders; Neuroglia; Nuclear; Oligodendroglia; PCNA gene; Pathway interactions; Patients; Plasmids; Play; Procedures; Process; Production; Progressive Multifocal Leukoencephalopathy; Proteins; PurA protein; Regulatory Pathway; Reporter Genes; Research Personnel; Role; Signal Pathway; Signal Transduction; Smad Proteins; Smad protein; T-Lymphocyte; Testing; Therapeutic immunosuppression; Tissues; Transgenic Organisms; Virus Activation; Virus Diseases; brain tissue; chromatin immunoprecipitation; cyclin T1; cytokine; in vivo; macrophage; monocyte; mouse model; programs; promoter; research study; tat Protein; viral DNA

DESCRIPTION (provided by applicant): The overall goal of this project is to elucidate the mechanisms by which activation of JC virus in glial cells of the brain is influenced by HIV-1 infection. JC virus (JCV) is the etiologic agent of the neurodegenerative disease, progressive multifocal leukoencephalopathy (PML). JCV is normally latent in non-immunocompromised people, but it is activated in brains of individuals with AIDS. Because of the high incidence of PML in AIDS, we have hypothesized that HIV-1 plays a role greater than that expected of immunosuppression alone. We shall capitalize upon our extensive results of the last few years, which can be grouped in two categories. In one we have demonstrated the role of the Tat protein of HIV-1 in stimulating JCV late gene transcription and JCV DMA replication. In the other we have demonstrated that HIV-1 infection alters pathways of production and signal transduction of immunomodulators, including TGF-01, in the CNS. We propose to determine how the Smad nuclear effectors of TGF-31 interact with Tat and its cellular partner proteins Pura and Cyclin T1/ Cdk9 at JCV and PCNA promoter sequences. Our new double chromatin immunoprecipitation method will be employed in conjunction with functional studies on JCV DMA replication and gene transcription. We shall employ a microarray to identify genes in glial cells regulated by exposure to cytokines produced by HIV-1-infected cells. We shall employ a transgenic mouse model to test the hypothesis that factors produced by HIV-1-infected cells can influence JCV promoters in the CNS and play a role in early steps of PML development. Tissue from the Manhattan HIV Brain Bank will be used to determine whether TGF-P1 or its nuclear effectors, Smad3, Smad4 or Fasti, are localized or activated in specific cells of PML lesions. Results will help elucidate pathways of activation of JCV in the brain and will help target particular molecular interactions for therapy.

描述（由申请人提供）：本项目的总体目标是阐明大脑神经胶质细胞中JC病毒活化受HIV-1感染影响的机制。JC病毒（JC virus，JCV）是神经退行性疾病--进行性多灶性白质脑病（Progressive Multifocal leukoencephalopathy，PML）的病原体。JCV通常潜伏在非免疫功能低下的人中，但它在艾滋病患者的大脑中被激活。由于艾滋病中PML的高发病率，我们假设HIV-1发挥的作用大于单独的免疫抑制。我们将利用过去几年取得的广泛成果，这些成果可分为两类。在一个实验中，我们已经证明了HIV-1的达特蛋白在刺激JCV晚期基因转录和JCV DMA复制中的作用。另一方面，我们已经证明HIV-1感染改变了CNS中免疫调节剂（包括TGF-01）的产生和信号转导途径。我们建议确定TGF-31的Smad核效应子如何与达特及其细胞伴侣蛋白Pura和Cyclin T1/Cdk 9在JCV和PCNA启动子序列处相互作用。我们新的双染色质免疫沉淀方法将与JCV DMA复制和基因转录的功能研究相结合。我们将采用微阵列来鉴定神经胶质细胞中受HIV-1感染细胞产生的细胞因子调节的基因。我们将采用转基因小鼠模型来检验HIV-1感染细胞产生的因子可以影响CNS中的JCV启动子并在PML发展的早期步骤中发挥作用的假设。来自曼哈顿HIV脑库的组织将用于确定TGF-β 1或其核效应物Smad 3、Smad 4或Fasti是否在PML病变的特定细胞中定位或活化。结果将有助于阐明JCV在大脑中的激活途径，并有助于靶向特定的分子相互作用进行治疗。