Molecular Analysis of the Ig Repertoire in MS

超出所提供的空间。多发性硬化（MS）是中枢神经系统（CNS）的炎性脱髓鞘疾病，其可能涉及针对自身髓鞘相关抗原的自身免疫机制。有大量证据表明，B细胞参与了至少一种MS发病机制。然而，克隆扩增的CSF B细胞产生的免疫球蛋白（IG）的功能和分子分析都受到限制。我们假设MS患者CSF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MS发病机制的至少一种。为了鉴定可能在MS患者中证明的自身免疫活性中发挥作用的完整IG重排并测试其抗原特异性，我们打算定义MS患者CSF中的IG谱系，以鉴定来自克隆扩增的B细胞的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重排的抗原特异性。我们还可以使用PCR来追踪这些克隆在原始MS患者中的持续性。此外，我们可以确定其他MS患者的CSF中是否具有相同的克隆扩增的B细胞。这些研究为启动和评估B细胞在MS发病机制中的至少一种机制中的潜在作用提供了必要的基础，最终将使我们能够探索将来是否有必要针对这些克隆扩增的B细胞产生特异性免疫疗法。性能现场=

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===========================================