1887

Abstract

(HCV) persists in the majority of infected individuals and is a major cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. Chronic hepatitis C is currently treated with interferon (IFN)-α or with a combination of IFN-α and ribavirin. The availability of an HCV replicon system (Lohmann , 285, 110–113, 1999) allowed the investigation of the effects of IFN on genuine HCV replication in cultured cells. It is shown here that IFN-α inhibits subgenomic HCV RNA replication in HuH-7 human hepatoma cells. Immunofluorescence, Western blot and Northern blot analysis revealed that levels of both HCV protein and replicon RNA were reduced after treatment with IFN-α in a dose-dependent manner. In further experiments, it was investigated whether MxA plays a role in the inhibition of HCV. The human MxA protein is an IFN-induced GTPase that has antiviral activity against various RNA viruses. However, HCV RNA replication was not affected in transfected HuH-7 cells that transiently overexpressed MxA. Moreover, a dominant-negative mutant of MxA did not interfere with the antiviral activity of IFN-α against HCV RNA replication. Taken together, these results demonstrate that IFN-α inhibits HCV replicons via an MxA-independent pathway.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-82-4-723
2001-04-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/82/4/0820723a.html?itemId=/content/journal/jgv/10.1099/0022-1317-82-4-723&mimeType=html&fmt=ahah

References

  1. Aebi M., Fäh J., Hurt N., Samuel C. E., Thomis D., Bazzigher L., Pavlovic J., Haller O., Staeheli P. 1989; cDNA structures and regulation of two interferon-induced human Mx proteins. Molecular and Cellular Biology 9:5062–5072
    [Google Scholar]
  2. Albanese M., Bruno-Smiraglia C., Di Cuonzo G., Lavagnino A., Srihongse S. 1972; Isolation of Thogoto virus from Rhipicephalus bursa ticks in western Sicily. Acta Virologica 16:267
    [Google Scholar]
  3. Antonelli G., Simeoni E., Turriziani O., Tesoro R., Redaelli A., Roffi L., Antonelli L., Pistello M., Dianzani F. 1999; Correlation of interferon-induced expression of MxA mRNA in peripheral blood mononuclear cells with the response of patients with chronic active hepatitis C to IFN-α therapy. Journal of Interferon and Cytokine Research 19:243–251
    [Google Scholar]
  4. Arnheiter H., Frese M., Kambadur R., Meier E., Haller O. 1996; Mx transgenic mice – animal models of health. Current Topics in Microbiology and Immunology 206:119–147
    [Google Scholar]
  5. Ausubel F. M., Brent R., Moore D. D., Seidman J. G., Smith J. A., Struhl K. (editors) 1997 Current Protocols in Molecular Biology New York: John Wiley;
  6. Bartenschlager R., Lohmann V. 2000; Replication of hepatitis C virus. Journal of General Virology 81:1631–1648
    [Google Scholar]
  7. Chomczynski P., Sacchi N. 1987; Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162:156–159
    [Google Scholar]
  8. Choo Q.-L., Kuo G., Weiner A. J., Overby L. R., Bradley D. W., Houghton M. 1989; Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359–362
    [Google Scholar]
  9. Davis G. L., Lau J. Y. 1997; Factors predictive of a beneficial response to therapy of hepatitis C. Hepatology 26:Suppl. 1122S–127S
    [Google Scholar]
  10. Enomoto N., Sakuma I., Asahina Y., Kurosaki M., Murakami T., Yamamoto C., Izumi N., Marumo F., Sato C. 1995; Comparison of full-length sequences of interferon-sensitive and resistant hepatitis C virus 1b. Sensitivity to interferon is conferred by amino acid substitutions in the NS5A region. Journal of Clinical Investigation 96:224–230
    [Google Scholar]
  11. Enomoto N., Sakuma I., Asahina Y., Kurosaki M., Murakami T., Yamamoto C., Ogura Y., Izumi N., Marumo F., Sato C. 1996; Mutations in the nonstructural protein 5A gene and response to interferon in patients with chronic hepatitis C virus 1b infection. New England Journal of Medicine 334:77–81
    [Google Scholar]
  12. Fernández M., Quiroga J. A., Martín J., Herrero M., Pardo M., Horisberger M. A., Carreño V. 1999; In vivo and in vitro induction of MxA protein in peripheral blood mononuclear cells from patients chronically infected with hepatitis C virus. Journal of Infectious Diseases 180:262–267
    [Google Scholar]
  13. Foster G. R., Thomas H. C. 2000; Therapeutic options for HCV – management of the infected individual. Ballière’s Best Practice and Research. Clinical Gastroenterology 14:255–264
    [Google Scholar]
  14. Frese M., Kochs G., Meier-Dieter U., Siebler J., Haller O. 1995; Human MxA protein inhibits tick-borne Thogoto virus but not Dhori virus. Journal of Virology 69:3904–3909
    [Google Scholar]
  15. Frese M., Kochs G., Feldmann H., Hertkorn C., Haller O. 1996; Inhibition of bunyaviruses, phleboviruses, and hantaviruses by human MxA protein. Journal of Virology 70:915–923
    [Google Scholar]
  16. Haller O., Frese M., Kochs G. 1998; Mx proteins: mediators of innate resistance to RNA viruses. Revue Scientifique et Technique Office International des Epizooties 17:220–230
    [Google Scholar]
  17. Hefti H. P., Frese M., Landis H., Di Paolo C., Aguzzi A., Haller O., Pavlovic J. 1999; Human MxA protein protects mice lacking a functional alpha/beta interferon system against La Crosse virus and other lethal viral infections. Journal of Virology 73:6984–6991
    [Google Scholar]
  18. Hijikata M., Ohta Y., Mishiro S. 2000; Identification of a single nucleotide polymorphism in the MxA gene promoter (G/T at nt −88) correlated with the response of hepatitis C patients to interferon. Intervirology 43:124–127
    [Google Scholar]
  19. Hoofnagle J. H. 1997; Hepatitis C: the clinical spectrum of disease. Hepatology 26:Suppl. 115S–20S
    [Google Scholar]
  20. Hoofnagle J. H., Mullen K. D., Jones D. B., Rustgi V., Di Bisceglie A., Peters M., Waggoner J. G., Park Y., Jones E. A. 1986; Treatment of chronic non-A, non-B hepatitis with recombinant human alpha interferon. A preliminary report. New England Journal of Medicine 315:1575–1578
    [Google Scholar]
  21. Jakschies D., Zachoval R., Müller R., Manns M., Nolte K. U., Hochkeppel H. K., Horisberger M. A., Deicher H., von Wussow P. 1994; Strong transient expression of the type I interferon-induced MxA protein in hepatitis A but not in acute hepatitis B and C. Hepatology 19:857–865
    [Google Scholar]
  22. Jones L. D., Nuttall P. A. 1989; The effect of virus-immune hosts on Thogoto virus infection of the tick, Rhipicephalus appendiculatus. Virus Research 14:129–139
    [Google Scholar]
  23. Kanerva M., Melen K., Vaheri A., Julkunen I. 1996; Inhibition of puumala and tula hantaviruses in Vero cells by MxA protein. Virology 224:55–62
    [Google Scholar]
  24. Keskinen P., Nyqvist M., Sareneva T., Pirhonen J., Melén K., Julkunen I. 1999; Impaired antiviral response in human hepatoma cells. Virology 263:364–375
    [Google Scholar]
  25. Korth M. J., Katze M. G. 2000; Evading the interferon response: hepatitis C virus and the interferon-induced protein kinase, PKR. Current Topics in Microbiology and Immunology 242:197–224
    [Google Scholar]
  26. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
    [Google Scholar]
  27. Landis H., Simon-Jödicke A., Kloti A., Di Paolo C., Schnorr J. J., Schneider-Schaulies S., Hefti H. P., Pavlovic J. 1998; Human MxA protein confers resistance to Semliki Forest virus and inhibits the amplification of a Semliki Forest virus-based replicon in the absence of viral structural proteins. Journal of Virology 72:1516–1522
    [Google Scholar]
  28. Leong J. C., Trobridge G. D., Kim C. H. Y., Johnson M., Simon B. 1998; Interferon-inducible Mx proteins in fish. Immunological Reviews 166:349–363
    [Google Scholar]
  29. Lohmann V., Körner F., Koch J. O., Herian U., Theilmann L., Bartenschlager R. 1999; Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line. Science 285:110–113
    [Google Scholar]
  30. Lohmann V., Körner F., Dobierzewska A., Bartenschlager R. 2001; Mutations in hepatitis C virus RNAs conferring cell culture adaptation. Journal of Virology 75:1437–1449
    [Google Scholar]
  31. McHutchison J. G., Gordon S. C., Schiff E. R., Shiffman M. L., Lee W. M., Rustgi V. K., Goodman Z. D., Ling M. H., Cort S., Albrecht J. K. 1998; Interferon alfa-2b alone or in combination with ribavirin as initial treatment for chronic hepatitis C. Hepatitis Interventional Therapy Group. New England Journal of Medicine 339:1485–1492
    [Google Scholar]
  32. MacQuillan G. C., de Boer W. B., Platten M. A., McCaul K. A., Reed W. D., Jeffrey G. P., Allan J. E. 2000; Intrahepatic MxA and PKR protein expression in chronic hepatitis C virus infection. Journal of Gastroenterology and Hepatology 15:Suppl.J56
    [Google Scholar]
  33. Marschall M., Zach A., Hechtfischer A., Foerst G., Meier-Ewert H., Haller O. 2000; Inhibition of influenza C viruses by human MxA protein. Virus Research 67:179–188
    [Google Scholar]
  34. Moradpour D., Blum H. E. 1999; Current and evolving therapies for hepatitis C. European Journal of Gastroenterology and Hepatology 11:1199–1202
    [Google Scholar]
  35. Nakabayashi H., Taketa K., Miyano K., Yamane T., Sato J. 1982; Growth of human hepatoma cells lines with differentiated functions in chemically defined medium. Cancer Research 42:3858–3863
    [Google Scholar]
  36. Pavlovic J., Zürcher T., Haller O., Staeheli P. 1990; Resistance to influenza virus and vesicular stomatitis virus conferred by expression of human MxA protein. Journal of Virology 64:3370–3375
    [Google Scholar]
  37. Pavlovic J., Haller O., Staeheli P. 1992; Human and mouse Mx proteins inhibit different steps of the influenza virus multiplication cycle. Journal of Virology 66:2564–2569
    [Google Scholar]
  38. Pavlovic J., Arzet H. A., Hefti H. P., Frese M., Rost D., Ernst B., Kolb E., Staeheli P., Haller O. 1995; Enhanced virus resistance of transgenic mice expressing the human MxA protein. Journal of Virology 69:4506–4510
    [Google Scholar]
  39. Pietschmann T., Lohmann V., Rutter G., Kurpanek K., Bartenschlager R. 2001; Characterization of cell lines carrying self-replicating hepatitis C virus RNAs. Journal of Virology 75:1252–1264
    [Google Scholar]
  40. Ponten A., Sick C., Weeber M., Haller O., Kochs G. 1997; Dominant-negative mutants of human MxA protein: domains in the carboxy-terminal moiety are important for oligomerization and antiviral activity. Journal of Virology 71:2591–2599
    [Google Scholar]
  41. Reed K. E., Rice C. M. 2000; Overview of hepatitis C virus genome structure, polyprotein processing, and protein properties. Current Topics in Microbiology and Immunology 242:55–84
    [Google Scholar]
  42. Robertson B., Myers G., Howard C., Brettin T., Bukh J., Gaschen B., Gojobori T., Maertens G., Mizokami M., Nainan O., Netesov S., Nishioka K., Shin-i T., Simmonds P., Smith D., Stuyver L., Weiner A. 1998; Classification, nomenclature, and database development for hepatitis C virus (HCV) and related viruses: proposals for standardization. International Committee on Virus Taxonomy. Archives of Virology 143:2493–2503
    [Google Scholar]
  43. Schneider-Schaulies S., Schneider-Schaulies J., Schuster A., Bayer M., Pavlovic J., ter Meulen V. 1994; Cell type-specific MxA-mediated inhibition of measles virus transcription in human brain cells. Journal of Virology 68:6910–6917
    [Google Scholar]
  44. Schnorr J. J., Schneider-Schaulies S., Simon-Jödicke A., Pavlovic J., Horisberger M. A., ter Meulen V. 1993; MxA-dependent inhibition of measles virus glycoprotein synthesis in a stably transfected human monocytic cell line. Journal of Virology 67:4760–4768
    [Google Scholar]
  45. Staeheli P., Haller O. 1985; Interferon-induced human protein with homology to protein Mx of influenza virus-resistant mice. Molecular and Cellular Biology 5:2150–2153
    [Google Scholar]
  46. Stark G. R., Kerr I. M., Williams B. R., Silverman R. H., Schreiber R. D. 1998; How cells respond to interferons. Annual Review of Biochemistry 67:227–264
    [Google Scholar]
  47. Taylor D. R. 2000; Hepatitis C virus: evasion of the interferon-induced antiviral response. Journal of Molecular Medicine 78:182–190
    [Google Scholar]
  48. Theodore D., Fried M. W. 2000; Natural history and disease manifestations of hepatitis C infection. Current Topics in Microbiology and Immunology 242:43–54
    [Google Scholar]
  49. Trepo C. 2000; Genotype and viral load as prognostic indicators in the treatment of hepatitis C. Journal of Viral Hepatitis 7:250–257
    [Google Scholar]
  50. Wölk B., Sansonno D., Kräusslich H. G., Dammacco F., Rice C. M., Blum H. E., Moradpour D. 2000; Subcellular localization, stability, and trans-cleavage competence of the hepatitis C virus NS3–NS4A complex expressed in tetracycline-regulated cell lines. Journal of Virology 74:2293–2304
    [Google Scholar]
  51. World Health Organization 1997; Hepatitis C: global prevalence. Weekly Epidemiological Record 72:341–344
    [Google Scholar]
  52. Zhao H., De B. P., Das T., Banerjee A. K. 1996; Inhibition of human parainfluenza virus-3 replication by interferon and human MxA. Virology 220:330–338
    [Google Scholar]
  53. Zhou A., Paranjape J. M., Der S. D., Williams B. R., Silverman R. H. 1999; Interferon action in triply deficient mice reveals the existence of alternative antiviral pathways. Virology 258:435–440
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-82-4-723
Loading
/content/journal/jgv/10.1099/0022-1317-82-4-723
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error