ENDOGENOUS RETROVIRUSES AS A CAUSAL MECHANISM IN LUPUS

内源性逆转录病毒(ERV)是人类和人类的潜在病原体。 小鼠自身免疫力。ERV的一类Mpmv在猪的胸腺中转录。 5只易患狼疮的小鼠，而11个对照品系中无一只。有狼疮倾向的人 小鼠，胸腺Mpmv RNA表达从出生第一天起就异常， 这表明它可能与易感基因密切相关。 自身免疫力。确定Mpmv直接编码的蛋白质 有助于自身免疫，它们的表达将在转基因中被阻止 狼疮鼠。我们的初步数据和发表的报告来自四个 针对其他基因(包括逆转录病毒)的不同组织表明 反义RNA对MPMV基因表达的调控作用 转基因小鼠。使用高表达Mpmv RNA和 蛋白质，我们将测试多个表达反义RNA的结构 不同启动子和增强子下不同的MPMV序列。 首先，我们将重点介绍以前使用的基于CMV的载体 成功在转基因小鼠中保护免受传染性逆转录病毒感染 通过表达反义RNA。由确定的最有希望的载体 将稳定的转基因T细胞系用于产生 转基因狼疮小鼠，然后将进行研究，以确定是否 这种疾病的表型已经改变了。 之前使用重组近交系和回交小鼠进行的遗传学研究已经 证明了几个自身免疫特征是独立分离的。至 确定胸腺Mpmv RNA表达是否可能与 自身免疫的特殊特征，我们将进行Northern分析 狼疮易感的NZB和NZB的重组自交系 对照组：SM/J小鼠。 最后，本申请将继续我们的初步研究， 提示Mpmv转录是基因调控失调的结果 在狼疮中的表达。引起MPMV异常的DNA序列 红斑狼疮易感小鼠的转录将被识别和表征。 由于狼疮的异常基因调控还不是很清楚，对 对Mpmv表达的调节可能会导致对Mpmv基因表达的新认识 人类和小鼠狼疮的分子遗传缺陷。

Endogenous retroviruses (ERV) are potential etiologic agents in human and murine autoimmunity. A class of ERV, Mpmv, is transcribed in thymuses of 5 lupus-prone but in none of 11 control mouse strains. In lupus-prone mice, thymus Mpmv RNA expression is abnormal from day one of life, suggesting that it may be closely linked to genes which predispose to autoimmunity. To determine whether the proteins encoded by Mpmv directly contribute to autoimmunity, their expression will be blocked in transgenic lupus mice. Our preliminary data and published reports from four different groups targeting other genes (including a retrovirus) indicate the potential utility of antisense RNA for manipulating Mpmv expression in transgenic mice. Using a T cell line that highly expresses Mpmv RNA and protein, we will test multiple constructs expressing antisense RNA to different Mpmv sequences under different promoters and enhancers. Initially, we will focus on the CMV-based vector previously used successfully in transgenic mice to protect against infectious retrovirus by expressing antisense RNA. The most promising vectors identified by stable transfection into the T cell line will be used to generate transgenic lupus-prone mice, which will then be studied to determine if the disease phenotype has been altered. Previous genetic studies using recombinant inbred and backcross mice have demonstrated that several autoimmune traits segregate independently. To determine whether thymus Mpmv RNA expression may cosegregate with a particular feature of autoimmunity, we will perform Northern analyses of well-characterized recombinant inbred lines between lupus-prone NZB and control SM/J mice. Finally, the present application will pursue our preliminary studies which indicate that Mpmv transcription results from a dysregulation of gene expression in lupus. The DNA sequences responsible for abnormal Mpmv transcription in lupus-prone mice will be identified and characterized. Since abnormal gene regulation in lupus is not well understood, studies of the regulation of Mpmv expression could lead to new insights into the molecular genetic defects of human and murine lupus.