CYTOKINE GENE EXPRESSION BY VIRUS IN GLIAL CELLS

描述：（申请人摘要）：我们的长期研究目标仍然是 同样：了解炎症和免疫介导的机制 中枢神经系统（CNS）损伤。 我们特别感兴趣 免疫效应物、细胞因子和趋化因子是如何在细胞内产生的。 中枢神经系统区室及其在中枢神经系统组织损伤中的作用。 这 应用的重点是研究神经营养性 病毒诱导神经胶质细胞中细胞因子和趋化因子基因表达， 研究TNF和RANTES基因。 促炎细胞因子TNF是一种有效的 原发性脱髓鞘效应。 RANTES，一种趋化因子， 细胞、单核细胞和树突状细胞在EAE脑中表达， 与疾病活动有关。 最近有报道称RANTES抑制 HIV在CD 4 + T细胞中的复制，这一发现可能与 CD 4 + T细胞对HIV暴露但未感染的相对抵抗力 个体 我们将使用纽卡斯尔病病毒（NDV）和麻疹病毒 （MV）。 我们以前用NDV研究过转录和mRNA的稳定性 的细胞因子和MV将是特别有用的描绘 RANTES基因的受体-配体诱导和dsRNA介导的信号传导 诱导，因为MV受体CD 46最近已被定义。 先前 我们实验室的研究表明，NDV诱导多种 细胞因子/趋化因子在原代大鼠星形胶质细胞和小胶质细胞中的表达。 我们也 显示MV在人胶质瘤细胞系和人原发性胶质瘤细胞系中诱导RANTES， 单核细胞 本研究拟探讨：（1）病毒诱导TNF的机制 通过鉴定稳定RNA来稳定星形胶质细胞中的mRNA 序列，表征RNA结合蛋白，并研究其作用 病毒诱导的激酶活性的mRNA稳定性。 (2)的机制 病毒诱导的RNATES基因在神经胶质细胞中的转录将通过 定义星形胶质细胞中RANTES启动子的病毒应答元件（VRE） 以及转录所需的VRE结合蛋白。 (3)信令 神经胶质细胞中MV诱导的RANTES基因表达所需的 通过定义MV对CD 46受体偶联的要求进行检查， RANTES基因表达所需的细胞内信号传导，使用大鼠 星形胶质细胞表达野生型CD 46或CD 46-GPI嵌合受体， 转染 此外，我们还将探讨RANTES的能力， 抑制星形胶质细胞中MV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.