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Susceptibility of human liver cell cultures to hepatitis C virus infection

  • G. Carloni
  • S. Iacovacci
  • M. Sargiacomo
  • G. Ravagnan
  • A. Ponzetto
  • C. Peschle
  • M. Battaglia
Part of the Archives of Virology Supplementum book series (ARCHIVES SUPPL, volume 8)

Summary

To develop a cell culture system susceptible to infection by hepatitis C virus (HCV), human fetal hepatocytes, grown in serum-free medium, were inoculated with serum samples from two HCV-infected patients. Viral RNA sequences were detected by polymerase chain reaction, using primers specific for the 5′ noncoding region of HCV, in extracts prepared from the hepatocyte cultures as early as 5 days after inoculation. Virus was also released from the infected cells into the medium. The HCV strains could be serially passaged three times into fresh liver cell cultures using intracellular virus as inoculum. Evidence that HCV replication really took place in primary human fetal hepatocytes was also obtained by detection of minus-strand viral RNA (replication intermediate) in cell extracts and of viral antigens in the infected cells.

Keywords

Hepatocyte Culture Human Liver Cell Fetal Liver Cell Liver Cell Culture Fetal Hepatocyte 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Choo QL, Kou G, Weiner AJ, Overby LR, Bradley DW, Houghthon M (1969) Isolation of cDNA clone from a blood-borne non-A, non-B viral hepatitis genome. Science 244: 359–361CrossRefGoogle Scholar
  2. 2.
    Esteban JI, Gonzalez A, Hernandez JM, Viladomiu L, Sanchez C, Lopez-Tavalera JC, Lucea D, Martin-Vega C, Vidal X, Esteban R, Guardia J (1990) Evaluation of antibodies to hepatitis C virus in a study of transfusion-associated hepatitis. N Engl J Med 323: 1107–1112PubMedCrossRefGoogle Scholar
  3. 3.
    Fong TL, Shindo M, Feinstone SM, Hoofnagle JH, Di Bisceglie AM (1991) Detection of replicative intermediate of hepatitis C viral RNA in liver and serum of patients with chronic hepatitis C. J Clin Invest 88: 1058–1060PubMedCrossRefGoogle Scholar
  4. 4.
    Ganem D, Varmus HE (1987) Molecular biology of the hepatitis viruses. Annu Rev Biochem 56: 651–693PubMedCrossRefGoogle Scholar
  5. 5.
    Garson JA, Tedder RS, Briggs M, Tuke A, Parker D, Barbara JAJ, Contreras M, Aloysius A (1990) Detection of hepatitis C viral sequences in blood donations by “nested” polymerase chain reaction and prediction of infectivity. Lancet 325: 1419–1422CrossRefGoogle Scholar
  6. 6.
    Gripon P, Diot C, Thézé N, Fourel I, Loreal O, Brechot C, Guguen-Guillouzo C (1988) Hepatitis B virus infection of adult human hepatocytes cultured in the presence of dimethyl sulfoxide. J Virol 62: 4136–4143PubMedGoogle Scholar
  7. 7.
    Jacob JR, Burk KH, Eichberg JW, Dreesman GR, Lanford RE (1990) Expression of infectious viral particles by primary chimpanzee hepatocytes isolated during the acute phase of non-A, non-B hepatitis. J Infect Dis 161: 1121–1127PubMedCrossRefGoogle Scholar
  8. 8.
    Kawasaki ES, Clark SS, Coyne MY, Smith SD, Champlin R, Witte ON, McCormick FP (1988) Diagnosis of chronic myeloid and acute lymphocytic leukemias by detection of leukemia-specific mRNA sequences amplified in vitro. Proc Natl Acad Sci USA 85: 5698–5702PubMedCrossRefGoogle Scholar
  9. 9.
    Lanford RE, Carey KD, Estlack LE, Smith GC, Hay B (1989) Analysis of plasma protein and lipoprotein synthesis in long-term primary cultures of baboon hepatocytes maintained in serum-free medium. In Vitro Cell Dev Biol 25: 174–182PubMedCrossRefGoogle Scholar
  10. 10.
    Moshage H, Yap SH (1992) Primary cultures of human hepatocytes: a unique system for studies in toxicology, virology parasitology and liver pathophysiology in man. J Hepatol 15: 404–411PubMedCrossRefGoogle Scholar
  11. 11.
    Nalpas B, Thiers V, Pol S, Driss F, Thepot V, Berthelot P, Brechot C (1992) Hepatitis C viremia and anti-HCV antibodies in alcoholics. J Hepatol 14: 381–384PubMedCrossRefGoogle Scholar
  12. 12.
    Novati R, Thiers V, D’Arminio Monforte A, Maisonneuve P, Principi N, Conti M, Lazzarin A, Brechot C (1992) Mother-to-child transmission of hepatitis C virus detected by nested polymerase chain reaction. J Infect Dis 165: 720–723PubMedCrossRefGoogle Scholar
  13. 13.
    Ochiya T, Tsurimoto T, Ueda K, Okubo K, Shiozawa M, Matsubara K (1989) An in vitro system for infection with hepatitis virus that uses primary human fetal hepatocytes. Proc Natl Acad Sci USA 86: 1875–1879PubMedCrossRefGoogle Scholar
  14. 14.
    Salas-Prato M, Tanguay JF, Lefebvre Y, Wojciechowicz D, Liem HH, Barnes DW, Ouellette G, Muller-Eberhard U (1988) Attachment and multiplication, morphology and protein production of human fetal primary liver cells cultured in hormonally defined media. In Vitro Cell Dev Biol 24: 230–238PubMedCrossRefGoogle Scholar
  15. 15.
    Tiollais P, Pourcel C, Dejean A (1985) The hepatitis B virus. Nature 317: 489–492PubMedCrossRefGoogle Scholar
  16. 16.
    Van der Poel CL, Cuypers HTL, Reesink HW, Weiner AJ, Quan S, Di Nello R, Van Boyen JJP, Winkel I, Mulder-Folkerts D, Exel-Oehlers PJ, Schaasberg W, Leentvaar-Kuypers A, Polito A, Houghton M, Lehe PN (1991) Confirmation of hepatitis C virus infection by new four-antigen recombinant immunoblot assay. Lancet 337: 317–319PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • G. Carloni
    • 1
  • S. Iacovacci
    • 1
  • M. Sargiacomo
    • 2
  • G. Ravagnan
    • 1
  • A. Ponzetto
    • 3
  • C. Peschle
    • 2
  • M. Battaglia
    • 1
  1. 1.Institute of Experimental MedicineIMS CNRRomeItaly
  2. 2.Laboratory of Hematology-OncologyInstituto Superiore di SanitàRomeItaly
  3. 3.Division of GastroenterologyLe Molinette HospitalTurinItaly

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