Molecular Analysis of the Ig Repertoire in MS

超出提供的空间。多发性硬化(MS)是一种中枢神经系统(CNS)炎症性脱髓鞘疾病，可能涉及针对自身髓鞘相关抗原的自身免疫机制。大量证据表明，B细胞参与了MS的至少一种发病机制。然而，对克隆扩增的脑脊液B细胞产生的免疫球蛋白(Ig)的功能和分子分析都是有限的。我们假设MS患者脑脊液中克隆扩增的B细胞亚群通过产生与髓鞘相关抗原结合的抗体参与了MS发病的至少一种机制。为了确定在MS患者自身免疫活动中可能起作用的完整的Ig重排，并检测其抗原特异性，我们打算确定MS患者脑脊液中的Ig谱系，以便从克隆性扩增的B细胞中识别Ig。然后，我们计划通过将重链和轻链片段克隆到表达载体中，分离得到的Fab片段，并测试它们的抗原特异性，来确定这些独特的Ig重排的抗原性。我们还可以使用聚合酶链式反应来追踪这些克隆在原始MS患者中的持久性。此外，我们可以确定其他MS患者的脑脊液中是否有相同的克隆性扩增的B细胞。这些研究为启动和评估B细胞在至少一种MS发病机制中的潜在作用提供了必要的基础，并最终将使我们能够探索未来是否有必要针对这些克隆扩增的B细胞进行特异性免疫治疗。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED. Multiple Sclerosis (MS) is an inflammatory, demyelinating disease of the Central Nervous System (CNS) which likely involves an autoimmune mechanism directed against self-myelin associated antigens. There is substantial evidence suggesting that B cells are involved in at least one mechanism of MS pathogenesis. However, both functional and molecular analysis of the immunoglobulins (Ig's) produced by clonally expanded CSF B cells has been limited. We hypothesize that a subset of clonally expanded B cells in the CSF of MS patients is involved in at least one mechanism of MS pathogenesis by producing antibodies that bind to myelin associated antigens. In order to identify complete Ig rearrangements that may play a role in the autoimmune activities evidenced in MS patients and test for their antigenic specificity, we intend to define the Ig repertoires in the CSF of MS patients in order to identify Ig's from clonally expanded B cells. We then plan to determine the antigenic specificity of these unique Ig rearrangements by cloning both the heavy and light chain segments into an expression vector, isolating the resultant Fab fragments, and testing them for their antigenic specificity. We can also use PCR to track the persistence of these clones in the original MS patients. Moreover, we can determine if other MS patients have the same clonally expanded B cells in their CSF. These studies provide the necessary foundation to initiate and assess the potential role of B cells in at least one mechanism of MS pathogenesis, and ultimately will allow us to explore whether generating specific immunotherapies directed against these clonally expanded B cells in the future is warranted. PERFORMANCE SITE ========================================Section End===========================================