N2a细胞增殖的痒病小鼠适应毒株RML的致病特性分析

doi:10.11843/j.issn.0366-6964.2015.02.012

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (2): 257-263.doi: 10.11843/j.issn.0366-6964.2015.02.012

N2a细胞增殖的痒病小鼠适应毒株RML的致病特性分析

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*

（1.扬州大学兽医学院，扬州 225009；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，青岛 266032）

收稿日期:2014-10-28 出版日期:2015-02-23 发布日期:2015-02-23
通讯作者: 王志亮，研究员，Tel：0532-85639166，E-mail：wangzhiliang@cahec.cn
作者简介:吴晓东（1975-），男，江苏宿迁人，副研究员，主要从事重大外来动物疫病的综合防控技术研究，E-mail：wuxiaodong@cahec.cn
基金资助:
转基因生物新品种培育科技重大专项（2014ZX08008-001B）

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

WU Xiao-dong^1，2，ZOU Yan-li²，LIU Yu-tian²，LI Jin-ming²，ZHANG Yong-qiang²，LIU Shan²，WANG Zhi-liang^2*

(1.School of Veterinary Medicine，Yangzhou University，Yangzhou 225009，China；2.National BSE Reference Lab，National Research Center for Exotic Animal Diseases，China Animal Health and Epidemiology Center，Qingdao 266032，China)

Received:2014-10-28 Online:2015-02-23 Published:2015-02-23

摘要/Abstract

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PrP^Sc与发病小鼠脑组织中PrP^Sc具有同样的糖基化模式，三种糖基化PrP^Sc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PrP^Sc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PrP^Sc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。

Abstract:

To identify the pathogenic characteristics of mouse-adapted Scrapie Strain RML propagated in N2a cell line，we intracerebrally inoculated mice with the lysate of Sc-N2a cell culture.After 200±28 days’ incubation period，all inoculated C57BL/6J mice appeared typical ataxia symptoms of TSEs.The PrP^Sc from both brain tissues and Sc-N2a cells 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 PrP^Sc deposits mainly in cortex，hippocampus，thalamus，cerebellum.The results of mouse regression test showed that the PrP^Scpropagated in N2a cell line had pathogenicity，and kept the the stain characteristics of mouse-adapted scrapie strain RML.The Sc-N2a cell line is an ideal model for the study of TSEs.

中图分类号:

S852.659.7

吴晓东，邹艳丽，刘雨田，李金明，张永强，刘珊，王志亮. N2a细胞增殖的痒病小鼠适应毒株RML的致病特性分析[J]. 畜牧兽医学报, 2015, 46(2): 257-263.

WU Xiao-dong，ZOU Yan-li，LIU Yu-tian，LI Jin-ming，ZHANG Yong-qiang，LIU Shan，WANG Zhi-liang. Pathogenic Characteristics of Mouse-adapted Scrapie Strain RML Propagated in N2a Cell Line[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(2): 257-263.

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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

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