人SCARB2蛋白提高人肠道病毒71型对转基因小鼠的感染力  

Human SCARB2 enhanced the infection of human enterovirus 71 on transgenic mice

作　　者：修晶辉[1] 刘江宁[1] 杨亚军[1] 夏咸柱 张连峰[1] 

机构地区：[1]医学实验动物研究所,中国医学科学院,北京协和医学院比较医学中心,卫生部人类疾病比较医学重点实验室,国家中医药管理局人类疾病动物模型三级实验室,北京100021 [2]军事医学科学学院,军事兽医研究所,吉林省人畜共患病防治重点实验室,吉林省长春市130122

出　　处：《中国比较医学杂志》2013年第7期16-20,I0002,共6页Chinese Journal of Comparative Medicine

基　　金：国家科技支撑计划课题(2012BA139B02);国家“重大新药创新”科技重大专项课题(课题编号2011ZX09307-302)

摘　　要：目的细胞实验证实人清道夫受体B2(hSCARB2)是人类EV71的受体,本研究拟通过建立人SCARB2转基因小鼠,建立感染能力更高的小鼠模型。方法构建CMV启动子的SCARB2转基因表达载体,通过显微注射法建立C57BL/6J背景的人SCARB2转基因小鼠,PCR筛选阳性首建鼠。通过Western Blot和免疫组化检测目的蛋白在各组织中的表达。EV71感染转基因小鼠后,通过实时荧光定量PCR和免疫组化检测目的蛋白表达对病毒感染的促进效果。结果人SCARB2蛋白主要在转基因小鼠的骨骼肌和脑组织中表达,与野生型小鼠相比,转基因小鼠组织中的病毒载量显著提高4到5倍。结论人SCARB2的体内表达可促进EV71对转基因小鼠的感染,该蛋白在体内具有EV71受体功能。Objective Human SCARB2 was identified as the functional receptors of EV71 in vitro, so establish more higher infection ability transgenic mice by establishment of hunman SCARB2 transgenic mice in present study. Methods The transgenic vector was constructed by inserting the human SCARB2 gene under the CMV promoter and then were subjected to establish transgenic mice by mieroinjection. The genotype of transgenic line was identified by PCR and the expression level of target protein was detected by Western blot. Viral load in the tissues of transgenic mice was detected by immunohistochemical staining and quantitative real-time PCR. Results One line of transgenic mice in C57BL/6J background with high levels of SCARB2 expression in skeletal muscle and brain was identified. Upon infection with EV71, the virus burden of 4 to 5 times in muscle and brain of transgenic mice were significantly higher than that of wild type mice. Conclusion hSCARB2 is a functional receptor of EV71 in vivo, as expression of it could promote the infection of EVT1 on transgenic mice.

关 键 词：人肠道病毒71型 人SCARB2蛋白 转基因小鼠 病毒受体 

分 类 号：R-332[医药卫生]