Hepatitis C virus genotype 1a growth and induction of autophagy.
We have previously reported that immortalized human hepatocytes (IHH) support the generation of infectious hepatitis C virus (HCV) genotype 1a (clone H77). In the present study, we have investigated the growth of HCV genotype 1a (clone H77) through serial passages and accompanying changes in IHH in response to infection. Eleven serial passages of HCV genotype 1a (clone H77) in IHH were completed. Virus replication was ascertained from the presence of HCV-specific sequences, the detection of core antigen, the virus genome copy number, and the virus titer in IHH culture fluid. Electron microscopy suggested that HCV infection induces autophagic vacuole formation in IHH. Fluorescence microscopy displayed localization of autophagic markers, microtubule-associated protein-1 light chain-3 and Apg5, on the vacuoles of HCV-infected hepatocytes. Taken together, our results suggested that HCV genotype 1a (clone H77) can be serially passaged in IHH and that HCV infection induces an autophagic response in hepatocytes.
Ait-Goughoulte
Malika
M
Division of Infectious Diseases and Immunology, Center for Vaccine Development, Edward A. Doisy Research Center, 1100 S. Grand Blvd., 8th Floor, St. Louis, MO 63104, USA.
Kanda
Tatsuo
T
Meyer
Keith
K
Ryerse
Jan S
JS
Ray
Ratna B
RB
Ray
Ranjit
R
D001343:Autophagy; D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D005838:Genotype; D006518:Hepatitis Viruses / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D008854:Microscopy, Electron; D016133:Polymerase Chain Reaction; D012692:Serial Passage
D017931:DNA Primers