Dynamic behavior of hepatitis C virus quasispecies in a long-term culture of the three-dimensional radial-flow bioreactor system.
Hepatitis C virus (HCV) exists in infected individuals as quasispecies, usually consisting of a dominant viral isolate and a variable mixture of related, yet genetically distinct, variants. A prior HCV infection system was developed using human hepatocellular carcinoma cells cultured in the three-dimensional radial-flow bioreactor (RFB), in which the cells retain morphological appearance and their differentiated hepatocyte functions for an extended period of time. This report studies the selection and alteration of the viral quasispecies in the RFB system inoculated with pooled serum derived from HCV carriers. Monitoring the viral RNA and core protein in the culture supernatants, together with nucleotide sequencing of hypervariable region 1 of the HCV genome, demonstrated that (1) the virus production intermittently fluctuated in the cultures, (2) the viral genetic diversity was markedly reduced 3 days post-infection (p.i.), and (3) dominant species changed on days 19-33p.i., suggesting that the virus populations can be selected according to susceptibility to the viral infection and replication. A therapeutic effect of interferon-alpha also demonstrated the inhibition of HCV expression. Thus, this HCV infection model in the RFB system should be useful for investigating the dynamic behavior of HCV quasispecies in cultured cells and evaluating anti-HCV compounds.
Murakami
Kyoko
K
Department of Virology II, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
Inoue
Yasushi
Y
Hmwe
Su-Su
SS
Omata
Kazuhiko
K
Hongo
Tomokatsu
T
Ishii
Koji
K
Yoshizaki
Sayaka
S
Aizaki
Hideki
H
Matsuura
Tomokazu
T
Shoji
Ikuo
I
Miyamura
Tatsuo
T
Suzuki
Tetsuro
T
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution / Q000235:genetics; D019149:Bioreactors / Q000821:virology*; D045744:Cell Line, Tumor; D014644:Genetic Variation; D016174:Hepacivirus / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D012367:RNA, Viral / Q000235:genetics; D012641:Selection, Genetic; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014758:Viral Core Proteins / Q000235:genetics; D014776:Virus Cultivation / Q000379:methods*
D012367:RNA, Viral; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus