ANTI-IDIOTYPIC ANTIBODIES IN MYASTHENIA

这个项目的总体目标是分析自身免疫反应。 重症肌无力(MG)患者乙酰胆碱受体(AchR)的表达及其意义 实验性自身免疫性重症肌无力(EAMG)模型； 来操纵自身抗体的表达。这一分析将 为开发抗原特异性或独特位奠定了基础 (ID)针对MG(以及其他自身免疫性疾病)的治疗 这将比目前可用的更有效，毒性更低 治疗。T细胞和B细胞免疫原性的系统研究 EAMG敏感株和耐药株AChR全基因表位分析 以确定T细胞库在小鼠体内的作用。 将进行抗体反应的多样性。T细胞 从T细胞克隆中定向使用受体(TCR)V区基因片段 针对免疫原性T细胞表位将通过血清学确定 并通过对这些细胞克隆的cDNA或基因组DNA进行测序。后者 研究将确定限制TCR基因片段的使用 在T细胞介导的自身免疫性疾病中观察到实验性变态反应 脑脊髓炎，也发生在抗体介导、T细胞控制 EAMG患者的自身免疫反应。此外，对EAMG的阻断效果 将评估针对重要TCR的抗克隆反应的程度。 T细胞在预防注射所致EAMG中的作用 将对先前开发的抗ID单抗进行分析。 这些研究将涉及从受保护的T细胞过继转移 动物对幼稚动物和TCR基因片段使用的测定 受保护的动物。拟议的实验将主要利用 肽扫描法检测T细胞和B细胞的AChR表位 免疫原性，T细胞克隆和杂交瘤的发展，扩增 用聚合酶链式反应和直接测序 以确定TCR基因片段的使用。

The overall goal of this project is to analyze the autoimmune response to the acetylcholine receptor (ACHR) in myasthenia gravis (MG) and its experimental model, experimental autoimmune myasthenia gravis (EAMG), and to manipulate the expression of the autoantibodies. This analysis will provide the groundwork for the development of antigen-specific, or idiotope (Id)-specific, treatments of MG (and other autoimmune diseases as well) that will be more effective and less toxic than presently available therapies. A systematic survey of both the T cell and B cell immunogenic epitopes of the entire AChR in EAMG-susceptible and EAMG-resistant strains of mice in order to determine the role of the T cell repertoire in the diversity of the antibody response will be carried out. The T cell receptor (TcR) V region gene segment usage from T cell clones directed against the immunogenic T cell epitopes will be determined serologically and by sequencing cDNA or genomic DNA from these cell clones. The latter studies will determine whether the restricted TcR gene segment usage observed in the T-cell-mediated autoimmune disease, experimental allergic encephalomyelitis, also occurs in the antibody-mediated, T-cell-controlled autoimmune response in EAMG. Moreover, the effectiveness in blocking EAMG of anti-clonotypic responses against important TcRs will be assessed. Also, the role of T cells in the protection from EAMG induced by injection of, previously developed, anti-Id monoclonal antibodies will be analyzed. These studies will involve adoptive transfer of T cells from protected animals to naive animals and determination of TcR gene segment usage in the protected animals. The proposed experiments will primarily make use of the pepscanning procedure to survey the AChR epitopes for T cell and B cell immunogenicity, development of T cell clones and hybridomas, amplification of cDNA and genomic DNA by polymerase chain reaction, and direct sequencing of amplified products to determine TcR gene segment usage.