N2a细胞增殖的痒病小鼠适应毒株RML的致病特性分析  

Pathogenic Characteristics of Mouse-adapted Scrapie Strain RML Propagated in N2a Cell Line

作　　者：吴晓东[1,2] 邹艳丽[2] 刘雨田[2] 李金明[2] 张永强[2] 刘珊[2] 王志亮[2] 

机构地区：[1]扬州大学兽医学院 [2]中国动物卫生与流行病学中心国家外来动物疫病研究中心国家BSE参考实验室

出　　处：《畜牧兽医学报》2015年第2期257-263,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：转基因生物新品种培育科技重大专项(2014ZX08008-001B)

摘　　要：为鉴定细胞体外增殖的痒病小鼠适应毒株RML的致病特性,包括致病性和株系特征等,用RML感染的N2a(Sc-N2a)细胞裂解物接种C57BL/6J小鼠进行研究。结果显示:经(200±28)d的潜伏期后,C57BL/6J小鼠全部发病死亡,临床症状均表现为共济失调,而接种N2a细胞裂解物的C57BL/6J小鼠与接种Sc-N2a细胞裂解物的PrP基因敲除小鼠均正常存活。Sc-N2a细胞增殖的PrPSc与发病小鼠脑组织中PrPSc具有同样的糖基化模式,三种糖基化PrPSc具相同的迁移率,而且均是以单糖基化形式为主。发病小鼠的病理学研究结果显示:脑部的海绵状空泡主要分布于脑干、大脑、小脑、丘脑;脑部的PrPSc主要分布于大脑皮质、海马、丘脑,在小脑、四迭体、嗅球有少量的分布。小鼠回归试验表明:Sc-N2a细胞增殖的PrPSc对小鼠具致病性,并保留RML的株系特征,是TSEs研究的理想细胞模型。To identify the pathogenic characteristics of mouse-adapted Scrapie Strain RML propagated in N2 acell line,we intracerebrally inoculated mice with the lysate of Sc-N2 acell culture.After 200±28days' incubation period,all inoculated C57BL/6Jmice appeared typical ataxia symptoms of TSEs.The PrPSc from both brain tissues and Sc-N2 acells shared the same biochemical aspects.Western blotting result confirmed that they had the same glycosylation patterns,the same size distribution of PK digestion fragments,with higher levels of monoglycosylated.The pathological changes of brain sections from inoculated mice at the terminal stage were analyzed by HE stain and IHC assay.Vacuoles were mainly found in brain stem,cortex,cerebellum and thalamus,while PrPSc deposits mainly in cortex,hippocampus,thalamus,cerebellum.The results of mouse regression test showed that the PrPSc propagated in N2 acell line had pathogenicity,and kept the the stain characteristics of mouse-adapted scrapie strain RML.The Sc-N2 acell line is an ideal model for the study of TSEs.

关 键 词：N2A细胞 痒病小鼠适应毒株RML PRPSC 致病性 株系特征 

分 类 号：S855.3[农业科学—临床兽医学]