Transgenic expression of Theiler's Virus encoded regions

该项目的目标是研究免疫反应的性质和特异性，这种免疫反应保护耐药小鼠免受泰勒氏小鼠脑脊髓炎病毒(TMEV)的持续感染，但也可能导致易感小鼠脱髓鞘和神经功能障碍。在这个多发性硬化症的小鼠模型中，免疫系统的功能不仅是清除病毒感染，而且还加剧了介导髓鞘和轴突损伤的致病反应。需要检验的假设是，TMEV基因组编码的抗原对保护性免疫(抵抗力)至关重要，但也可能导致免疫病理学(易感性)。这些实验将利用一系列表达TMEV基因组三个独立连续区域的转基因小鼠。转基因小鼠是在TMEV基因组的I类连续区域控制下产生的。在表达VP1I区编码序列5‘(L、VP4、VP2和VP3)、区II(VP1编码块)和区III编码序列3’(VP92A、2B、2C、3A、3B、3C和3D)的I类启动子的控制下创建了转基因小鼠。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对TMEV编码区的免疫反应的作用。这些实验还将评估感染了TMEV的表达人类II类MHC基因的转基因小鼠的脱髓鞘和神经功能缺陷。最后，我们将利用穿孔素缺陷小鼠的过继转移实验，研究导致神经功能缺陷的免疫反应的表型和特异性。当注射TMEV时，小鼠显示脱髓鞘，但不显示神经功能障碍。这些实验有望为髓鞘损伤和与人类多发性硬化症相关的神经缺陷的机制提供独特的见解。

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.