髓鞘少突胶质细胞糖蛋白MOG抗原肽免疫原性验证系列.doc

ＸＸ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 PAGE \* MERGEFORMAT 3 PAGE \* MERGEFORMAT I 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MOG35~55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： MOG35~55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MOG35~55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 Abstract Objective The 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). Methods Each 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. Results The incidence of EAE in immunization animal reached 100%, most mice developed clinical EAE with in about 15 days of immunization MOG35~55 peptides. With Electron microscopy, inflammatory cells in white matter and demyelination were observed. And the component of axon is disappeared. Conclusion The immunogenicity of MOG35~55 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