was基因缺陷小鼠mcmv急性感染模型的建立及感染特点.doc

WAS基因缺陷小鼠MCMV急性感染模型的建立及感染特点 张玉琳 罗小华 刘 林 （重庆医科大学附属第一医院血液内科，重庆，400016） [摘要] 目的 用小鼠巨细胞病毒（MCMV）Smith株建立WAS基因缺陷小鼠（129S6/SvEvTac-Wastm1Sbs/J）全身播散型感染模型并观察感染特点。方法 同窝内简单随机法选取实验组小鼠（129S6/SvEvTac） WASp-/-（雌雄各半），感染对照组小鼠（129S1/SvImNJ）WASp+/+ 6只，雌雄各半，68周龄。腹腔内接种0.2m小鼠巨细胞病毒悬液1×105 PFU；另设6只129S6小鼠为空白对照组。各组分别接种MCMV 9后处死小鼠，取小鼠唾液腺分离病毒，RTPCR检测小鼠主要脏器组织中MCMV gB基因，HE病理染色观察小鼠脏器病理损害，流式细胞术检测WAS基因缺陷小鼠脾脏MCMV特异性细胞毒性T细胞（CTL）比例。结果 与对照组野生型小鼠相比，实验组WASp-/-小鼠MCMV感染后一般状况差，体质量下降，活动少，耸毛明显，有死亡（1/6）。感染后第9天，实验组和感染对照组小鼠唾液腺均分离出巨细胞病毒，主要脏器心、肝、肺、肾和唾液腺中MCMV gB基因RTPCR检测结果为阳性。实验组小鼠肺内MCMV gB基因mRNA含量显著高于对照组小鼠（P0.05），空白对照组未检测出病毒。MCMV感染后主要脏器出现明显病理损害，其中肺部病理损害严重。流式结果显示WAS基因缺陷小鼠脾脏MCMV CTL比率与对照组无差别（P=0.88）。结论 首次腹腔注射MCMV 1×105 PFU建立成年WAS基因缺陷小鼠急性感染模型，观察到较对照组野生型小鼠更明显的、较重的急性感染反应；肺为感染主要靶器官之一；初次感染后脾脏MCMV CTL比例无明显变化。 [关键词] 鼠巨细胞病毒；WAS基因缺陷小鼠；感染模型；流式细胞术 [中图法分类号] R559 R-332 [文献标志码] A Establishment of WASp-deficient mice model of acute murine cytomegalovirus infection and infection characteristics Zhang Yulin, Luo Xiaohua, Liu Lin (Department of Hematology, First Affiliated Hospital, Chongqing Medical University, Chongqing, 400016, China) [Abstract] Objective To establish a WASp-deficient mouse (129S6/SvEvTac-Wastm1Sbs/J) model of induced systemic infection with murine cytomegalovirus (MCMV) and investigate the infection characteristics. Methods WAS-null mice were randomly chosen as model group and blank control group respectively (n=6 for each group). They were infected with 1×105 PFU MCMV (0.2 mL) by intra-peritoneal administration. WASp+/+ mice were used as infection control (n=6). All experimental mice were under closely observation and sacrificed in 9 d after the intraperitoneal injection. Tissue samples were collected under aseptic conditions from each experimental mouse. Salivary glands were taken for virus separation and titer test. The histological changes of the main visceral organs and tissues were examined after HE staining, and the expression of MCMV gB was detected by RT