HepG2 cells support viral replication and gene expression of hepatitis C virus genotype 4 in vitro.
OBJECTIVE
To establish a cell culture system with long-term replication of hepatitis C virus (HCV) genome and expression of viral antigens in vitro.


METHODS
HepG2 cell line was tested for its susceptibility to HCV by incubation with a serum from a patient with chronic hepatitis C. Cells and supernatant were harvested at various time points during the culture. Culture supernatant was tested for its ability to infect naive cells. The presence of minus (antisense) RNA strand, and the detection of core and E1 antigens in cells were examined by RT-PCR and immunological techniques (flow cytometry and Western blot) respectively.


RESULTS
The intracellular HCV RNA was first detected on d 3 after infection and then could be consistently detected in both cells and supernatant over a period of at least three months. The fresh cells could be infected with supernatant from cultured infected cells. Flow cytometric analysis showed surface and intracellular HCV antigen expression using in house made polyclonal antibodies (anti-core, and anti-E1). Western blot analysis showed the expression of a cluster of immunogenic peptides at molecular weights extended between 31 and 45 kDa in an one month old culture of infected cells whereas this cluster was undetectable in uninfected HepG2 cells.


CONCLUSIONS
HepG2 cell line is not only susceptible to HCV infection but also supports its replication in vitro. Expression of HCV structural proteins can be detected in infected HepG2 cells. These cells are also capable of shedding viral particles into culture media which in turn become infectious to uninfected cells.
el-Awady
Mostafa K
MK
Department of Biomedical Technology, National Research Center, Tahrir Street, PO 12622, Dokki, Cairo, Egypt. mkawady@yahoo.com
Tabll
Ashraf A
AA
el-Abd
Yasmine S
YS
Bahgat
Mahmoud M
MM
Shoeb
Hussein A
HA
Youssef
Samar S
SS
Bader el-Din
Noha G
NG
Redwan
el-Rashdy M
el-RM
el-Demellawy
Maha
M
Omran
Moataza H
MH
el-Garf
Wael T
WT
Goueli
Said A
SA
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D003470:Culture Media / Q000737:chemistry; D015967:Gene Expression Regulation, Viral; D005838:Genotype; D016174:Hepacivirus / Q000502:physiology; D006801:Humans; D008113:Liver Neoplasms; D014779:Virus Replication
D000956:Antigens, Viral; D003470:Culture Media