IMMUNOPATHIC MYOSITIS INDUCED BY PERSISTENT VIRUS

持续病毒引起的免疫病性肌炎

• 批准号: 2072371
• 负责人: RONALD P MESSNER
• 金额: $ 17.57万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-30 至 1997-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2072371
• 关键词: Coxsackievirus; RNase protection assay; artificial immunosuppression; athymic mouse; autoimmune disorder; computer assisted sequence analysis; denervation; disease /disorder model; genetic regulation; genetic strain; host organism interaction; laboratory mouse; latent virus infection; molecular cloning; myositis; natural selections; tissue /cell culture; virus RNA; virus diseases

Enterovirus infection has been linked to the expression of several human autoimmune diseases including polymyositis, juvenile-onset diabetes, and cardiomyopathy. Evidence of prior enterovirus infection includes elevated antibody titers against virus and the detection of persistent viral RNA in the afflicted tissue. However, results from some studies are equivocal, and hence the role of enterovirus in precipitating autoimmune diseases is still unclear. Techniques for the detection of persistent enteroviruses are hampered by a lack of knowledge about the mechanism which underlies persistence. Similarly, the mariner in which persistent enterovirus provokes the induction or maintenance of immunologically-mediated tissue damage is not understood. This proposal employs a mouse model of coxsackievirus B 1-induced polymyositis to study mechanisms of virus persistence and evaluate the effect of virus persistence on immunopathic muscle disease. Two key attributes of this model are that viral RNA persists for long periods of time in muscle, and immunopathic disease is mediated by T cells. The first objective is to explore the nature of persistent virus and determine whether it is comprised of altered forms of viral RNA and/or infectious virus. Nuclease protection assays will be used to identify genomic deletions, sequence changes, and altered ratios of positive and negative RNA strands. Reactivation of infectious virus will be attempted with several different strategies including immunosuppression and muscle denervation. A somewhat unique strategy for virus reactivation will be to culture muscle in vitro where it is removed from potential anti-viral effects of the immune system. Tissues other than muscle that have previously been found to harbor persistent viral RNA will also be tested as potential reservoirs of virus. If infectious virus is recovered, it will be phenotypically and genotypically characterized and compared with the original infecting strain. The second objective seeks to better understand factors that influence the coevolution of host and virus, and focuses on selective pressures exerted by the host on development of virus persistence. Nude mice and inbred strains of mice with varying susceptibilities to myositis will be used to examine the influence of T cells and host background genes on the establishment of persistent infection. The third objective is to identify muscle genes that are differentially regulated as a result of virus persistence. The differential display technique will be used to analyze muscle for RNA transcripts that are upregulated or downregulated at different stages of the disease. Taken together, these experiments will clarify important interactions between persistent virus and the host that culminate in the development of immunopathic muscle disease. They will also lead to more effective approaches for the epidemiologic study of persistent virus- induced autoimmune disease.

肠道病毒感染与几个人的表达有关 自身免疫性疾病，包括多发性肌炎、青少年发病的糖尿病和 心肌病。以前感染过肠道病毒的证据包括 猪瘟病毒抗体滴度及持久性病毒RNA的检测 受苦的组织。然而，一些研究的结果是模棱两可的， 因此，肠道病毒在诱发自身免疫性疾病中的作用是 仍不清楚。持久性肠道病毒的检测技术 由于缺乏对其基础机制的了解而受到阻碍 坚持不懈。同样，水手体内持续存在的肠道病毒 刺激免疫介导的组织的诱导或维持 损害还不能理解。这项提案使用了一种老鼠模型 柯萨奇病毒B_1诱导的多发性肌炎病毒机制的研究 病毒持久性及其对免疫病理影响的评价 肌肉疾病。该模型的两个关键属性是病毒RNA 在肌肉中持续很长一段时间，免疫疾病是 由T细胞介导。第一个目标是探索 持续病毒，并确定它是否由改变形式的 病毒RNA和/或传染性病毒。将使用核酸酶保护分析 确定基因组的缺失、序列改变和改变的比率 正负核糖核糖核酸链。传染性病毒的重新激活将 尝试几种不同的策略，包括免疫抑制 和肌肉失神经。一种有点独特的病毒重新激活策略 将是在体外培养肌肉，在那里它被从潜在的 免疫系统的抗病毒作用。肌肉以外的组织 以前被发现携带持久性病毒RNA的人也会 被检测为潜在的病毒宿主。如果传染性病毒被发现， 将对其进行表型和基因特征分析和比较 与原始的感染菌株相匹配。第二个目标是寻求更好地 了解影响宿主和病毒协同进化的因素，以及 重点关注宿主对病毒发展施加的选择性压力 坚持不懈。裸鼠和近交系小鼠具有不同的 对肌炎的易感性将被用来检查T的影响 细胞和宿主背景基因对持久性的建立 感染。第三个目标是识别肌肉基因 由于病毒的持久性而受到不同的监管。这个 差异显示技术将用于分析肌肉中的RNA 在不同阶段上调或下调的转录本 这种疾病。总而言之，这些实验将阐明重要的 持久性病毒与宿主之间的相互作用，最终导致 免疫性肌肉疾病的发展。它们还将带来更多 持续病毒流行病学研究的有效途径-- 诱发自身免疫性疾病。