AChRα亚基基因片段联合IL-6DNA免疫大鼠建立实验性重症肌无力模型  

Building EAMG models by DNA immunization combined gene encoding AChR alpha subunit with cDNA of IL-6

作　　者：孟和宝力高 郭素杰[1] 赵文双[1] 额尔敦[1] 

机构地区：[1]中国人民解放军第二五三医院免疫中心,呼和浩特010051

出　　处：《免疫学杂志》2010年第6期531-535,共5页Immunological Journal

摘　　要：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。The aim of the study is to build an experimental autoimmune myasthenia gravis （EAMG） animal model by DNA im- munization for further study. AChR α205, encoding the main immunogenic region of N-terminal 205 residues of human nicotinic acetylcholine receptor alpha 1 subunit, and rat interleukin-6 cDNA sequence were respectively subcloned into pcDNA3.1（＋）, for constructing pcDNA3.1（＋）/AChR α205 and pcDNA3.1（＋）/IL-6 recombinant plasmids. Female Lewis rats in control group were inoculated by pcDNA3.1（＋）; rats in two experimental groups were inoculated by pcDNA3.1（＋）/AChR α205 and pcDNA3.1（＋）/AChR α205 with pcDNA3.1 （＋）/IL-6, respectively. For all these mice, we investigated the clinical manifestation, electromyogram, anti-AChR antibody, histopathology and ultramicrostructure, gene integration, and RNA transcription. And we found that clinical muscle was weakened, repetitive nerve stimulation was attenuated, serum anti-AChR antibody was increased, and retrogression was appeared in histological examination as well as ultramicrostructure in a few rats of experimental groups. These changes were more obvious in rats inoculated by both of the two recombinant plasmids. Target gene was all integrated into genomes and RNA also transcribed in the two experimental groups. In our study, EAMG animal models were built by inoculated the two recombinant plasmids pcDNA3.1 （＋） /AChR α205 and pcDNA3.1（＋）/IL-6 to Lewis rats, which will be a convenient and practical way for building animal model.

关 键 词：重症肌无力 实验性自身免疫性重症肌无力 乙酰胆碱受体 DNA免疫 

分 类 号：R746.1[医药卫生—神经病学与精神病学]