Transgenic expression of Theiler's Virus encoded regions

泰勒病毒编码区的转基因表达

• 批准号: 6652309
• 负责人: MOSES RODRIGUEZ
• 金额: $ 19.52万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6652309
• 关键词: MHC class II antigen; cytolysins; disease /disorder model; genetic mapping; genetic promoter element; genetic susceptibility; genetically modified animals; histocompatibility gene; immunity; immunopathology; laboratory mouse; microorganism immunology; multiple sclerosis; murine encephalomyelitis virus; myelinopathy; nerve injury; neurologic manifestations; pore forming protein; virus genetics

The goal of this project is to investigate the nature and specificity of the immune response that protects resistance mice from Theiler's murine encephalomyelitis virus (TMEV) persistent infection but which may also contribute in susceptible mice to demyelination and neurologic deficits. In this murine model of multiple sclerosis the immune system functions both to clear virus infection but also to exacerbate the pathogenic response which mediates myelin and axonal injury. The hypothesis to be tested is that antigens encoded by the TMEV genome are critical for protective immunity (resistance) but possibility may also contribute to immunopathology (susceptibility). The experiments will utilize a series of transgenic mice expressing independent three continuous regions of the TMEV genome. Transgenic mice have been created under control of a class I continuous regions of the TMEV genome. Transgenic mice have been created under control of a class I promoter expressing region I coding sequence 5' of VP1 (L, VP4, VP2, and VP3), region II (VP1 coding block), and region III coding sequence 3' of VP1 92A, 2B, 2C, 3A, 3B, 3C, and 3D). By challenging mice expressing TMEV transgenes with infectious virus, we will be able to address the role of immune response to TMEV coding regions in vivo. These experiments will also evaluate demyelination and neurologic deficits in transgenic mice expressing human class II MHC genes infected with TMEV. Finally we will study the phenotype and specificity of the immune response contributing to neurologic deficits utilizing adoptive transfer experiments with perforin deficient mice when injected with TMEV show demyelination but fail to show neurologic deficits. The experiments are expected to provide unique insights into the mechanisms of myelin injury and neurologic deficits with relevance to human multiple sclerosis.

该项目的目标是研究免疫反应的性质和特异性，该免疫反应可以保护耐药小鼠免受泰勒氏鼠脑脊髓炎病毒（TMEV）持续感染，但也可能导致易感小鼠脱髓鞘和神经功能缺陷。在这种多发性硬化症小鼠模型中，免疫系统不仅可以清除病毒感染，还可以加剧介导髓磷脂和轴突损伤的致病反应。要测试的假设是 TMEV 基因组编码的抗原对于保护性免疫（抵抗性）至关重要，但可能性也可能有助于免疫病理学（易感性）。实验将利用一系列表达 TMEV 基因组的独立三个连续区域的转基因小鼠。转基因小鼠是在 TMEV 基因组 I 类连续区域的控制下创建的。转基因小鼠已在表达 VP1 5' 区 I 编码序列（L、VP4、VP2 和 VP3）、II 区（VP1 编码块）和 VP1 3' 区 III 编码序列（92A、2B、2C、3A、3B、3C 和 3D）的 I 类启动子控制下创建。通过用感染性病毒攻击表达 TMEV 转基因的小鼠，我们将能够解决体内 TMEV 编码区免疫反应的作用。这些实验还将评估表达感染 TMEV 的人类 II 类 MHC 基因的转基因小鼠的脱髓鞘和神经缺陷。最后，我们将利用穿孔素缺陷小鼠的过继转移实验，研究导致神经缺陷的免疫反应的表型和特异性，当注射 TMEV 时，小鼠表现出脱髓鞘，但未表现出神经缺陷。这些实验有望为与人类多发性硬化症相关的髓磷脂损伤和神经缺陷的机制提供独特的见解。