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Theiler小鼠脑脊髓炎病毒（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。在多发性硬化症小鼠模型中，免疫系统既能清除病毒感染，又能加剧介导髓鞘和轴索损伤的致病性反应。待验证的假设是，由TMEV基因组编码的抗原对保护性免疫（抗性）至关重要，但也可能有助于免疫病理（易感性）。该实验将利用一系列转基因小鼠表达独立的三个连续的TMEV基因组区域。转基因小鼠是在TMEV基因组的一类连续区域的控制下创建的。在一类启动子的控制下建立了转基因小鼠，该启动子表达VP1的I区编码序列5′（L、VP4、VP2和VP3）、VP1的II区（VP1编码块）和VP1的III区编码序列3′（2a、2B、2C、3A、3B、3C和3D）。通过用感染性病毒挑战表达TMEV转基因的小鼠，我们将能够在体内解决免疫应答对TMEV编码区的作用。这些实验还将评估表达人类II类MHC基因的转基因小鼠感染TMEV后脱髓鞘和神经功能缺陷。最后，我们将利用过继性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，穿孔蛋白缺陷小鼠出现脱髓鞘，但没有出现神经功能缺陷。该实验有望为髓磷脂损伤和与人类多发性硬化症相关的神经功能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.