大学毕业论文题 目：髓鞘少突胶质细胞糖蛋白 MOG 抗原肽免疫原性验证学生姓名： XXX 学号： XXXXXXXXX 指导教师： XXX 职称： 教授 专 业： 生物技术 院（系）： 生物工程系 完成时间： 2011 年 5 月 25 日 2011 年 5 月 25 日摘要目的：通过实验性自身免疫性脑脊髓炎（Experimental autoimmune encephalomyelitis, EAE）小鼠模型的建立，验证固相合成法制备的髓鞘少突胶质细胞糖蛋白（MOG）抗原肽免疫原性。并通过观察 EAE 病理学变化，初步探讨多发性硬化（MS ）的发病机制。方法：本实验室利用 Fmoc 固相合成法制备的 MOG3555 抗原肽（纯度95%）加完全弗氏佐剂（CFA ）免疫 C57BL/6 小鼠，观察小鼠发病情况，并采用双盲法进行神经功能评分。利用电子显微镜观察 EAE 小鼠的病理学变化，分别采用 HE 染色、Luxol Fast Blue髓鞘染色法评估脊髓中炎性细胞的浸润及髓鞘脱失的情况。结果：MOG3555 成功诱导了 C57BL/6 小鼠 EAE 模型。免疫后 15 天起，小鼠开始陆续出现临床症状，至第 21 天发病率达 100%。电子显微镜下可见 EAE 小鼠脊髓中典型脱髓鞘现象，并伴随有典型的炎性细胞的浸润。结论：用本实验室合成的抗原肽 MOG3555 诱导的 C57BL/6 小鼠 EAE 模型，发病率高，模型稳定，为进一步研究多发性硬化（multiple sclerosis, MS）的发病机制与治疗打下基础。关键词：髓鞘少突胶质细胞糖蛋白、实验性自身免疫性脑脊髓炎、动物模型、神经病理学IAbstractObjectiveThe model of experimental autoimmune encephalomyelitis(EAE) was established in female C57BL/6 mice to validate the immunogenicity of myelin oligodendrocyte glycoprotein (MOG), to observe pathological changes of EAE, and to explore the possible pathogenesis of multiple sclerosis(MS).MethodsEach mouse was injected s.c over flanks with MOG35-55, Mycobacterium H37Ra, and CFA, injected i.p with pertussis toxin. The clinical symptoms were observed and the pathological changes of EAE were studied with the electron microscopy. Consecutive sections were stained with hematoxylin/eosin(HE) and Luxol Fast Blue(LFB), to assess inflammation and demyelination.ResultsThe incidence of EAE in immunization animal reached 100%, most mice developed clinical EAE with in about 15 days of immunization MOG3555 peptides. With Electron microscopy, inflammatory cells in white matter and demyelination were observed. And the component of axon is disappeared.ConclusionThe immunogenicity of MOG3555 synthesized by the improved solid-phase synthesis method was identified by the induction of EAE in C57BL/6 mice with stable performance and high incidence, which may be helpful in exploring the mechanism and therapy of Multiple sclerosis(MS). Key Words: Myelin oligodendrocyte glycoprotein(MOG), Experimental autoimmune encephalomyelitis(EAE), Mouse models, Pathological changesIIIII目录引言 .1一、材料和方法 .21. 试剂和仪器 .21.1. 主要 试剂 .21.2 主要仪器 .22. 实验动物 .22.1 小鼠来源 .22.2 饲养 .33. EAE 小鼠模型的制备 .34. 神经功能评分 .35. 脊髓组织病理学检测取材 .36. 溶液的配制 .37. H&E、LFB 染色 .47.1 H&E 染色 .47.2 LFB 染色 .48. 数据分析及统计学处理 .5二、实验结果与分析 .51. 临床评分 .