CYTOKINE GENE EXPRESSION BY VIRUS IN GLIAL CELLS

描述：(申请人摘要)：我们的长期研究目标保持不变 同样：了解炎症和免疫介导的机制 中枢神经系统损伤。我们特别感兴趣的是 免疫效应器、细胞因子和趋化因子是如何在 以及它们在中枢神经系统组织损伤中的作用。这 应用重点是研究嗜神经性的机制 病毒通过以下途径诱导胶质细胞细胞因子和趋化因子基因表达 研究肿瘤坏死因子和RANTES基因。促炎细胞因子肿瘤坏死因子是一种有效的 初级脱髓鞘的效应器。RANTES，一种吸引T细胞的趋化因子 细胞、单核细胞和树突状细胞，在EAE脑中表达，以及 与疾病活动性相关。兰特最近被报道抑制了 HIV在CD4+T细胞中的复制，这一发现可能与 CD4+T细胞对HIV暴露但未感染的相对抵抗力 个人。我们将使用新城疫病毒(NDV)和麻疹病毒 (Mv)。我们以前使用新城疫病毒来研究转录和mRNA的稳定性 细胞因子和MV将特别有用，以描绘 RANTES基因受体配体诱导及dsRNA介导的信号转导 诱导，因为最近定义了MV受体CD46。上一首 我们实验室的研究表明，新城疫病毒可以诱导多种不同的 原代大鼠星形胶质细胞和小胶质细胞中的细胞因子/趋化因子。我们也 结果显示MV可诱导人脑胶质瘤细胞系和人原代细胞株RANTES 单核细胞。我们想要研究：(1)病毒诱导的肿瘤坏死因子的机制 通过鉴定稳定的RNA来稳定星形胶质细胞的mRNA 测序，鉴定RNA结合蛋白，并研究其作用 病毒诱导的蛋白激酶活性在信使核糖核酸的稳定性。(2)机制问题。 病毒诱导的胶质细胞RNAES基因转录将通过以下方式进行探索 星形胶质细胞RANTES启动子病毒反应元件的确定 以及转录所需的VRE结合蛋白。(3)信令 MV诱导的胶质细胞RANTES基因表达所需的 通过定义MV和CD46受体偶联的要求进行检查 RANTES基因表达所需的细胞内信号转导 表达CD46或CD46-GPI嵌合受体的星形胶质细胞 转染法。此外，我们还将探索RANTES的能力 抑制星形胶质细胞中MV的复制。

DESCRIPTION: (Applicant's abstract): Our long term research goal remains the same: to understand the mechanisms of inflammation and immune-mediated injury of the central nervous system (CNS). We are specifically interested in how immune effectors, cytokines and chemokines are generated within the CNS compartment and what role they play in CNS tissue injury. This application focuses on investigating the mechanisms by which neurotropic viruses induce cytokine and chemokine gene expression in glial cells by studying TNF and RANTES genes. Pro-inflammatory cytokine TNF is a potent effector in primary demyelination. RANTES, a chemokine that attracts T cells, monocytes and dendritic cells, is expressed in EAE brains, and correlates with disease activity. RANTES was recently reported to inhibit HIV replication in CD4+ T cells, a finding that may be related to the relative resistance of CD4+ T cells to HIV-exposed but uninfected individuals. We will use Newcastle Disease virus (NDV) and measles virus (MV). We used NDV previously to study the transcription and mRNA stability of cytokines and MV will be especially useful to delineate the receptor-ligand induced, and dsRNA-mediated signaling for RANTES gene induction, because the MV receptor CD46 has been recently defined. Previous studies in our laboratory have shown that NDV induces a variety of cytokines/chemokines in primary rat astrocytes and microglia. We also showed that MV induces RANTES in a human glioma cell line and human primary monocytes. We want to investigate: (1) The mechanisms of virus-induced TNF mRNA stabilization in astrocytes by identifying the stabilizing RNA sequence, characterize the RNA-binding proteins, and investigating the role of virus-induced kinase activity in mRNA stability. (2) The mechanisms of virus-induced RNATES gene transcription in glial cells will be explored by defining the virus response elements (VRE) of RANTES promoter in astrocytes and the VRE-binding proteins required for the transcription. (3) Signaling required for MV-induced RANTES gene expression in glial cells will be examined by defining the requirement of CD46 receptor coupling by MV and intracellular signaling required for RANTES gene expression, using rat astrocytes expressing wild CD46 or CD46-GPI chimeric receptors through transfection. In addition, we will also explore the ability of RANTES to inhibit MV replication in astrocytes.