Observation of chimeric hepatitis C virus In infected Huh7.5 cell through transmission electron microscopy
10.3760/cma.j.issn.0254-5101.2009.06.020
- VernacularTitle:体外培养的嵌合体丙型肝炎病毒感染Huh7.5细胞的透射电镜观察
- Author:
Li MA
;
Xin WEI
;
Ye ZHANG
;
Pingzhong WANG
;
Jianqi LIAN
;
Zhansheng JIA
- Publication Type:Journal Article
- Keywords:
Hepatitis C virus;
In vitro transcription;
Transmission electron microscopy
- From:
Chinese Journal of Microbiology and Immunology
2009;29(6):568-573
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the morphological characteristics of HCV particles and intracel-lular ultrastructure changes in Huh7. 5 cells which was infected with chimeric HCV via transmission electron microscopy. Methods Plasmid J6/JFH encoding the full length HCV chimeric genome was transcribed to HCV RNA in vitro and the RNA was transfected into Huh7.5 cells by electroporation. Quantitative real-time PCR(qRT-PCR) was used to assay HCV copies of the supernatant of transfected cells. Indirect immunofluo-rescence was used to detect the expression of HCV proteins. The cell culture superoatant were used to infect narve Huh7.5 cells, transmission electron microscopy was used to observe morphological characteristics of vi-rus particles and intracellular ultrastructure changes in infected Huh7. 5 cells. Results qRT-PCR showed high level virus copies in supernatant of transfected cells collected in different times, indirect immuno-fluo-rescencc proved high expression of HCV NS5A proteins in the transfected cells. Large numbers of enveloped or unenveloped virus-like particles (VLPs) were observed in infected Huh7. 5 cells via transmission electronmicroscopy. We also found hyperplasia of some membrane-enclosed organelles in the cytoplasm. Several fea-tures characterizing flavivirus infected cells and a cytoplasmic inclusion of unknown origin were observed. Conclusion The chimeric HCV from in vitro cell culture system is proved to be intact virus particles which can efficiently infect Huh7.5 cells.
