摘要
目的运用不同的技术方法,观察SIVmac239感染CEMx174细胞的全过程。方法 SIVmac239病毒感染CEMx174细胞后,分别在不同的时间点收集培养上清和细胞,应用实时荧光定量PCR方法检测培养上清中的病毒载量;IFA检测并用激光扫描共聚焦显微镜观察病毒定位及复制情况;胞内流式、Western blot检测细胞内病毒蛋白p27的表达量。结果 SIVmac239吸附并感染CEMx174后,第0.5 h时,细胞胞膜上检测到病毒颗粒;第12h时,培养上清中的病毒RNA水平出现下降;此后至第96 h,病毒持续复制,细胞胞浆内病毒蛋白p27表达量逐渐增加,培养上清中病毒RNA水平持续升高。结论病毒感染细胞时,首先吸附在细胞膜上,然后进入细胞持续复制和表达。
Objective To observe the process of SIVmac239 adsorbing onto and infecting target cell line CEMx174 and revealing the basis for the virus entering target cells.Methods CEMx174 cells were infected by the SIVmac239 virus.Culture supernatants and cells were collected at various time points.The viral load in culture supernatants was detected by quantitative real-time PCR.Expression of intracellular viral protein p27 was detected by indirect immunofluorescence,intracellular staining,and Western blotting.Results CEMx174 cells absorbed the virus and were infected.Virus particles were detected on the cell membrane at 0.5 h.At 12 h,the level of viral RNA in the culture supernatant was reduced.Then up to 96 h,the expression of SIV p27 in the cytoplasm was increased,and the level of viral RNA in the culture supernatants increased continually.Conclusions When the virus infects its target cells,it is first adsorbed through virus-associated receptors and then enters the cell.
作者
李想
童玲
丛喆
薛婧
LI Xiang;TONG Ling;CONG Zhe;XUE Jing(Institute of Laboratory Animal Sciences,Chinese Academy of Medical Sciences(CAMS);Comparative Medicine Center,Peking Union Medical College(PUMC);Key Laboratory of Human Disease Comparative Medicine,National Health and Family Planning Commission of the People’s Republic of China(NHFPC);Key)
出处
《中国比较医学杂志》
CAS
北大核心
2018年第7期7-11,共5页
Chinese Journal of Comparative Medicine
基金
中国医学科学院医学与健康科技创新工程重大协同创新项目(编号:2017-I2M-1-014)
科技部重大专项(编号:2017ZX10304402)。
