SIVmac239 病毒感染CEMx174 细胞过程分析

doi:10.3969/j. issn. 1671 -7856. 2018. 07. 002

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SIVmac239 病毒感染CEMx174 细胞过程分析
DOI:
                        10.3969/j. issn. 1671 -7856. 2018. 07. 002
                    
作者:
                        李想李想
中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,卫计委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,新发再发传染病动物模型研究北京市重点实验室,北京 100021
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童玲童玲
中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,卫计委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,新发再发传染病动物模型研究北京市重点实验室,北京 100021
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丛喆丛喆
中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,卫计委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,新发再发传染病动物模型研究北京市重点实验室,北京 100021
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薛婧薛婧
中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,卫计委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,新发再发传染病动物模型研究北京市重点实验室,北京 100021
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作者单位:(中国医学科学院医学实验动物研究所,北京协和医学院比较医学中心,卫计委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,新发再发传染病动物模型研究北京市重点实验室,北京 100021)
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中图分类号:R-33
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The process of CEMx174 cell infection by SIVmac239

Author:

LI Xiang
LI Xiang
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 Laboratory of Human Diseases Animal Model, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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TONG Ling
TONG Ling
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 Laboratory of Human Diseases Animal Model, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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CONG Zhe
CONG Zhe
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 Laboratory of Human Diseases Animal Model, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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XUE Jing
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 Laboratory of Human Diseases Animal Model, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China
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Affiliation:

(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 Laboratory of Human Diseases Animal Model, State Administration of Traditional Chinese Medicine; Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious Diseases, Beijing 100021, China)

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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12 h 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?

关键词:猴免疫缺陷病毒;CEMx174 细胞;病毒蛋白p27;吸附;感染

Abstract:

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.

Key words:simian immunodeficiency virus, SIV; CEMx174 cells; viral protein p27; adsorption; infection

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李想,童玲,丛喆,薛婧. SIVmac239 病毒感染CEMx174 细胞过程分析[J].中国比较医学杂志,2018,28(7):7~11.

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收稿日期:2017-12-28
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在线发布日期: 2018-08-08
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