汉滩病毒感染小鼠腹腔巨噬细胞模型的建立  

Establishment of Hantaan Virus Infected Mouse Peritoneal Elicit Macrophage

作　　者：张尧[1] 冯伟[1] 张晓晓[2] 应旗康[2] 刘梓谕[2] 吴兴安[2] 王芳[2] 

机构地区：[1]第四军医大学学员旅,陕西西安710032 [2]第四军医大学微生物学教研室,陕西西安710032

出　　处：《生物技术通讯》2017年第4期503-505,共3页Letters in Biotechnology

基　　金：国家自然科学基金(31470890;31270978);军队重点课题(BWS13G);陕西省科技统筹创新工程计划(2013KTCL03-06)

摘　　要：目的:建立汉滩病毒感染的小鼠腹腔巨噬细胞模型。方法:PBS灌洗6~8周龄C57BL/6小鼠腹腔,分离获取小鼠腹腔巨噬细胞后,以100 TCID50汉滩病毒76-118株感染小鼠腹腔巨噬细胞,通过间接免疫荧光、ELISA和Realtime PCR检测病毒感染情况。结果:病毒感染3 d后,间接免疫荧光和ELISA检测到病毒核衣壳蛋白的表达,Realtime PCR检测到病毒核酸的表达。结论:建立了汉滩病毒感染小鼠腹腔巨噬细胞的模型,为进一步阐明汉滩病毒的发病机制奠定了基础。Objective： To establish a macrophage model for Hantaan virus（HTNV） infection by using the cul- ture of mouse peritoneal elicit macrophage. Methods： Inject 5 mL of ice cold PBS（with 3% FCS） into the peritoneal cavity. After injection, gently massage the peritoneum to dislodge any attached cells into the PBS solution and then collect as much as possible, and deposit the collected cell suspension in tubes on ice. The mouse peritoneal macrophages were cultured and infected with 100 TCIDs0 HTNV 76-118 strain. Indirect immunofluorescent assay（IFA）, ELISA and real-time PCR were used to detect the protein or nucleic acid of Hantaan virus. Results： 3 days after infection, the nucleoprotein was detected out by IFA and ELISA, the nucleic acid of nucleoprotein was confirmed by real-time PCR. Conclusion： Direct infection of HTNV on the cultured mouse peritoneal macrophages were identified, to explore the pathogenic mechanism for HTNV infection.

关 键 词：汉滩病毒 小鼠腹腔巨噬细胞 感染 

分 类 号：R373[医药卫生—病原生物学] R392.1[医药卫生—基础医学]