论文部分内容阅读
目的:探索用DNA免疫方法:建立实验性自身免疫性重症肌无力(EAMG)动物模型的可行性。方法:将含人乙酰胆碱受体α亚基N端主要免疫区205个氨基酸(AChRα205)的编码基因、含大鼠IL-6cDNA序列的基因分别亚克隆入真核表达载体pcDNA3.1(+)构建pcDNA3.1(+)/AChRα205、pcDNA3.1(+)/IL-6两种重组质粒。以pcDNA3.1(+)免疫对照组Lewis大鼠,pcDNA3.1(+)/AChRα205和pcDNA3.1(+)/AChRα205加pcDNA3.1(+)/IL-6分别免疫两个实验组大鼠。从临床症状、肌电图、血清抗AChR抗体、组织病理与超微结构、组织基因整合和RNA转录几方面进行检测与评估。结果:实验组动物出现了临床肌无力症状;重复神经电刺激呈衰减反应;血清抗AChR抗体滴度增高;组织学与超微结构检查呈退行性改变;这些变化在双质粒共注射组更明显。两实验组大鼠均有目的:基因的整合与转录。结论:用重组质粒pcDNA3.1(+)/AChRα205、pcDNA3.1(+)/IL-6免疫Lewis大鼠建立了EAMG模型,方法:简便实用。
Objective: To explore the feasibility of using DNA immunization method to establish an animal model of experimental autoimmune myasthenia gravis (EAMG). Methods: The gene encoding 205 amino acids (AChRα205), containing the rat IL-6 cDNA sequence, was subcloned into the eukaryotic expression vector pcDNA3.1 (+) pcDNA3.1 (+) / AChRα205, pcDNA3.1 (+) / IL-6 two recombinant plasmids. The Lewis rats, pcDNA3.1 (+) / AChRα205 and pcDNA3.1 (+) / AChRα205 plus pcDNA3.1 (+) / IL-6 were immunized with pcDNA3.1 (+ . From the clinical symptoms, electromyography, serum anti-AChR antibodies, histopathology and ultrastructure, tissue gene integration and RNA transcription aspects of detection and evaluation. Results: The mice in the experimental group showed symptoms of clinical myasthenia gravis. The repetitive electrical nerve stimulation showed an attenuated response. The serum anti-AChR antibody titer increased. Histological and ultrastructural examination showed degenerative changes. These changes were more obvious in the double plasmid co-injection group . Both experimental groups rats have the purpose of: gene integration and transcription. Conclusion: EAMG model was established by immunizing Lewis rats with recombinant plasmid pcDNA3.1 (+) / AChRα205 and pcDNA3.1 (+) / IL-6, and the method was simple and practical.