摘要
为检测钙离子、镁离子对戊型肝炎病毒(hepatitis E virus,HEV)感染PLC/PRF/5细胞(人肝癌亚历山大细胞)的影响,本研究在各实验组PLC/PRF/5细胞的培养体系中加入等量HEV毒种进行孵育,利用实时荧光定量反转录聚合酶链反应以及酶联免疫法,监测HEV核酸和抗原含量;在HEV感染细胞实验组的维持培养液中分别加入钙离子、镁离子、乙二胺四乙酸(ethylene diamine tetraacetic acid,EDTA)和乙二醇二乙醚二胺四乙酸(ethylene glycol tetraacetic acid,EGTA),观察钙离子、镁离子、EDTA和EGTA分别存在以及钙离子、镁离子同时存在的情况下,HEV感染后不同时间PLC/PRF/5细胞内及培养上清液中HEV含量。结果显示,HEV接种细胞后1~24h,钙离子、镁离子的加入能够促进病毒与细胞的结合,而金属离子鏊合剂的加入抑制了病毒与细胞的结合。HEV感染后2~5周,钙离子、镁离子均能增加PLC/PRF/5细胞培养上清液中产生的病毒,其中钙离子的促进作用更加显著。本研究结果表明钙离子、镁离子能够促进HEV感染细胞,在HEV接种细胞后的培养过程中,添加钙离子、镁离子有助于病毒产生。
To examine the effect of calcium and magnesium on hepatitis E virus(HEV)infection in PLC/PRF/5 cells,quantitative real-time fluorescent RT-PCR and ELISA were used to detect the viral nucleic acid and antigen content of HEV.The virus nucleic acid and ORF2 antigen in the cells and supernatant were examined at different time points after HEV infection in the presence of EGTA,calcium or magnesium respectively.The addition of calcium and magnesium to the cell culture medium increased virus binding,whereas chelating agents inhibited HEV binding 1 to 24 hours after HEV incubation.Both calcium and magnesium increased the production of HEV in the cell culture medium of PLC/PRF/5 cells after two to five weeks,while calcium had more pronounced effect than magnesium.Calcium and magnesium could promote HEV infection in PLC/PRF/5 cells.The addition of calcium and magnesium in the HEV culture is helpful for virus production.
作者
张黎
黄维金
ZHANG Li;HUANG Weijin(Division of HIV/AIDS and Sex-transmitted Virus Vaccines,National Institutes of Food and Drug Control,Beijing 102629,China)
出处
《微生物与感染》
CAS
2021年第3期171-175,共5页
Journal of Microbes and Infections
基金
国家自然科学基金(81371830)。