Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro

Silvia Iacovacci, Aldo Manzin, Stefano Barca, Massimo Sargiacomo, Annalucia Serafino, Maria Beatrice Valli, Giampiero Macioce, Hamina Jane Hassan, Antonio Ponzetto, Massimo Clementi, Cesare Peschle, Guido Carloni

Research output: Contribution to journalArticle

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Abstract

The molecular features of hepatitis C virus (HCV) replication in human fetal hepatocytes (HFHs) were addressed in this study. Using a competitive reverse-transcription polymerase chain reaction (RT-PCR) assay for the quantitation of HCVRNA molecules, the highest level of viral replication was detected 30 days' postinfection. At this time point, viral particles of 41 to 45 nm in diameter accumulated in the cell cytoplasm. Their density in cell extracts and culture medium was distributed between heavy (1.180-1.360 g/cm3) and light fractions (1.105-1.050 g/cm3) of a sucrose gradient, while, in the serum inoculum, they had a positive fraction at 1.180 g/cm3. In infected HFHs, minus-strand HCV RNA was observed in fractions displaying a sedimentation coefficient of 28 S to 18 S, while plus-strand HCV RNA showed a peak restricted to the 21 S fraction; the HCV RNA of serum inoculum had a sedimentation coefficient of 38 to 40 S, which revealed the presence of HCV RNA of unique positive polarity. The 21 S RNA fraction of cell extracts was resistant to 20 minutes of RNase I digestion, while the same incubation time totally inactivated a comparable amount of HCV RNA purified from the serum inoculum, revealing the presence of completely and/or partially double- stranded HCV-RNA molecules in the infected cells. Detection in HFHs of replicative forms and replicative intermediates suggests that the dynamic profile of HCV replication in these cells is similar to that described in other flaviviruses.

Original languageEnglish
Pages (from-to)1328-1337
Number of pages10
JournalHepatology
Volume26
Issue number5
Publication statusPublished - Nov 1997

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Molecular Dynamics Simulation
Virus Replication
Hepacivirus
Hepatocytes
RNA
Cell Extracts
Serum
Flavivirus
Pancreatic Ribonuclease
In Vitro Techniques
Virion
Reverse Transcription
Culture Media
Sucrose
Digestion
Cytoplasm
Cell Culture Techniques
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Hepatology

Cite this

Iacovacci, S., Manzin, A., Barca, S., Sargiacomo, M., Serafino, A., Valli, M. B., ... Carloni, G. (1997). Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro. Hepatology, 26(5), 1328-1337.

Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro. / Iacovacci, Silvia; Manzin, Aldo; Barca, Stefano; Sargiacomo, Massimo; Serafino, Annalucia; Valli, Maria Beatrice; Macioce, Giampiero; Hassan, Hamina Jane; Ponzetto, Antonio; Clementi, Massimo; Peschle, Cesare; Carloni, Guido.

In: Hepatology, Vol. 26, No. 5, 11.1997, p. 1328-1337.

Research output: Contribution to journalArticle

Iacovacci, S, Manzin, A, Barca, S, Sargiacomo, M, Serafino, A, Valli, MB, Macioce, G, Hassan, HJ, Ponzetto, A, Clementi, M, Peschle, C & Carloni, G 1997, 'Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro', Hepatology, vol. 26, no. 5, pp. 1328-1337.
Iacovacci S, Manzin A, Barca S, Sargiacomo M, Serafino A, Valli MB et al. Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro. Hepatology. 1997 Nov;26(5):1328-1337.
Iacovacci, Silvia ; Manzin, Aldo ; Barca, Stefano ; Sargiacomo, Massimo ; Serafino, Annalucia ; Valli, Maria Beatrice ; Macioce, Giampiero ; Hassan, Hamina Jane ; Ponzetto, Antonio ; Clementi, Massimo ; Peschle, Cesare ; Carloni, Guido. / Molecular characterization and dynamics of hepatitis C virus replication in human fetal hepatocytes infected in vitro. In: Hepatology. 1997 ; Vol. 26, No. 5. pp. 1328-1337.
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