中国实验动物学报2017，Vol.25Issue(2)：117-122,6.DOI:10.3969/j.issn.1005-4847.2017.02.002
EV71病毒对树鼩原代肾上皮细胞感染模型的建立
Establishment of a EV71 virus infection model of tree shrew primary renal cells
摘要
Abstract
Objective To establish an enterovirus 71(EV71) infection model of tree shrew primary renal cells.Methods Tree shrew primary renal cells were obtained by trypsin digestion.After subculture and purification,EV71 virus was used to infect these primary cells.The culture supernatant of these EV71-infected cells was collected for virus titer detection at 1,2,4,6 and 8 days post-infection.The cells were collected for detection of EV71 VP1 protein by Western blot assay.Furthermore,the expression and location of VP1 protein in the infected cells were detected by indirect immunofluorescence assay.Vero cells were taken as positive control to evaluate the infectivity of EV71 virus to tree shrew primary renal cells.Results Morphologically,the cultured cells were proved to be majorly consisted of the primary renal cells after subculture and purification.The obtained primary cells were infected by EV71 virus.The virus titer was up to 1.3×106 TCID 50/mL during 48-96 h post-infection,proving that EV71 virus infected and proliferated in the tree shrew primary renal cells.Western blot showed that the viral VP1 protein was detected from infected primary cells at 2 to 8 d post infection.VP1 protein was also observed in the cytoplasm at 2 to 6 d post infection by indirect immunofluorescence.Compared with Vero cells,the infectivity of EV71 virus to tree shrew primary renal cells and its proliferation were confirmed.Conclusions Based on the successful establishment of cell culture of tree shrew primary renal cells,the infectivity to the obtained cells and proliferation of EV71 virus in the cells are confirmed.The model of EV71 virus-infected tree shrew primary renal cells is initially established.关键词
树鼩原代肾细胞/EV71/病毒增殖/VP1蛋白/感染模型Key words
Tree shrew/primary renal cells/EV71 virus/Virus proliferation/Protein VP1/Infection model分类
生物科学引用本文复制引用
杨铭,胡晓星,王文广,张莉,董书维,冯悦,代解杰,夏雪山..EV71病毒对树鼩原代肾上皮细胞感染模型的建立[J].中国实验动物学报,2017,25(2):117-122,6.基金项目
国家科技支撑计划项目子课题(2014BAI01B01). （2014BAI01B01）
