Intervirology 2006;49:352–361
(DOI:10.1159/000095155)

Evolution of Hepatitis C Virus Quasispecies during Ribavirin and Interferon-Alpha-2b Combination Therapy and Interferon-Alpha-2b Monotherapy

Arataki K.a · Kumada H.b · Toyota K.c · Ohishi W.c · Takahashi S.c · Tazuma S.c · Chayama K.c
aDepartment of Internal Medicine, Kure Medical Association Hospital, Kure-shi, bDepartment of Gastroenterology, Toranomon Hospital, Tokyo, and cDepartment of Medicine and Molecular Science, Division of Frontier Medical Science, Programs for Biomedical Research, Graduate School of Biomedical Sciences, Hiroshima University, Minami-ku, Hiroshima, Japan
email Corresponding Author


 goto top of outline Key Words

  • Hepatitis C virus quasispecies
  • Viral resistance
  • Error catastrophe
  • Chronic hepatitis C virus infection
  • Ribavirin

 goto top of outline Abstract

Objective: Ribavirin and interferon combination therapy is more effective than interferon monotherapy in patients with chronic hepatitis C virus (HCV) infection. To test the hypothesis that ribavirin induces nucleotide substitutions in the viral genome and reduces viral load by forcing it into error catastrophe in the combination therapy, we investigated the molecular evolution of HCV quasispecies in 3 patients who received combination therapy and 2 patients who received interferon monotherapy. Methods: The quasispecies were analyzed before and after therapy by sequencing at least 8 clones in five regions of the HCV genome; 5′ untranslated region, EI, E2, NS5A and NS5B. Results: Marked genetic drift was observed in the NS5A and NS5B regions in patients treated with combination therapy. However, genetic distances between clones obtained after therapy were closer than those obtained before therapy. Conclusion: Our results suggest that the combination therapy modified HCV quasispecies, but that this did not reflect the induction of error catastrophe by ribavirin. Modification of quasispecies by this therapy requires further investigation in a larger number of patients to elucidate the possible mechanism of viral resistance against the combination therapy.

Copyright © 2006 S. Karger AG, Basel


 goto top of outline References
  1. Tong MJ, El-Farra NS, Reikes AR, Co RL: Clinical outcomes after transfusion-associated hepatitis. N Engl J Med 1995;332:1463–1466.
  2. Kim WR: The burden of hepatitis C in the United States. Hepatology 2002;36(suppl 1):S30–S34.

    External Resources

  3. Darby SC, Ewart DW, Giangrande PL, Spooner RJ, Rizza CR, Dusheiko GM, Lee CA, Ludlam CA, Preston FE: Mortality from liver cancer and liver disease in haemophilic men and boys in UK given blood products contaminated with hepatitis C. UK Haemophilia Centre Directors’ Organisation. Lancet 1997;350:1425–1431.
  4. Kiyosawa K, Tanaka E: Characteristics of hepatocellular carcinoma in Japan. Oncology 2002;62(suppl 1):5–7.
  5. McHutchison JG, Gordon SC, Schiff ER, Shiffman ML, Lee WM, Rustgi VK, Goodman ZD, Ling MH, Cort S, Albrecht JK: Interferon alfa-2b alone or in combination with ribavirin as initial treatment for chronic hepatitis C. Hepatitis Interventional Therapy Group. N Engl J Med 1998;339:1485–1492.
  6. Poynard T, Marcellin P, Lee SS, Niederau C, Minuk GS, Ideo G, Bain V, Heathcote J, Zeuzem S, Trepo C, Albrecht J: Randomised trial of interferon alpha2b plus ribavirin for 48 weeks or for 24 weeks versus interferon alpha2b plus placebo for 48 weeks for treatment of chronic infection with hepatitis C virus. International Hepatitis Interventional Therapy Group (IHIT). Lancet 1998;352:1426–1432.
  7. Davis GL, Esteban-Mur R, Rustgi V, Hoefs J, Gordon SC, Trepo C, Shiffman ML, Zeuzem S, Craxi A, Ling MH, Albrecht J: Interferon alfa-2b alone or in combination with ribavirin for the treatment of relapse of chronic hepatitis C. International Hepatitis Interventional Therapy Group. N Engl J Med 1998;339:1493–1499.
  8. Lau JYN, Tam RC, Liang TJ, Hong Z: Mechanism of action of ribavirin in the combination treatment of chronic HCV infection. Hepatology 2002;35:1002–1009.
  9. Crotty S, Maag D, Arnold JJ, Zhong W, Lau JY, Hong Z, Andino R, Cameron CE: The broad-spectrum antiviral ribonucleoside ribavirin is an RNA virus mutagen. Nat Med 2000;6:1375–1379.
  10. Crotty S, Cameron CE, Andino R: RNA virus error catastrophe: direct molecular test by using ribavirin. Proc Natl Acad Sci USA 2001;98:6895–6900.
  11. Contreras AM, Hiasa Y, He W, Terella A, Schmidt EV, Chung RT: Viral RNA mutations are region specific and increased by ribavirin in a full-length hepatitis C virus replication system. J Virol 2002;76:8505–8517.
  12. Zhou S, Liu R, Baroudy BM, Malcolm BA, Reyes GR: The effect of ribavirin and IMPDH inhibitors on hepatitis C virus subgenomic replicon RNA. Virology 2003;310:333–342.
  13. Sookoian S, Castano G, Frider B, Cello J, Campos R, Flichman D: Combined therapy with interferon and ribavirin in chronic hepatitis C does not affect serum quasispecies diversity. Dig Dis Sci 2001;46:1067–1071.
  14. Polyak SJ, McArdle S, Liu SL, Sullivan DG, Chung M, Hofgartner WT, Carithers RL Jr, McMahon BJ, Mullins JI, Corey L, Gretch DR: Evolution of hepatitis C virus quasispecies in hypervariable region 1 and the putative interferon sensitivity-determining region during interferon therapy and natural infection. J Virol 1998;72:4288–4296.
  15. Pawlotsky JM, Germanidis G, Frainais PO, Bouvier M, Soulier A, Pellerin M, Dhumeaux D: Evolution of the hepatitis C virus second envelope protein hypervariable region in chronically infected patients receiving alpha interferon therapy. J Virol 1999;73:6490–6499.
  16. Bassett SE, Thomas DL, Bransly KM, Lanford RE: Viral persistence, antibody to E1 and E2, and hypervariable region 1 sequence stability in hepatitis C virus-inoculated chimpanzees. J Virol 1999;73:1118–1126.
  17. Gale MJ Jr, Korth MJ, Tang NM, Tan SL, Hopkins DA, Dever TE, Polyak SJ, Gretch DR, Katze MG: Evidence that hepatitis C virus resistance to interferon is mediated through repression of the PKR protein kinase by the nonstructural 5A protein. Virology 1997;230:217–227.
  18. Pawlotsky JM, Germanidis G, Neumann AU, Pellerin M, Frainais PO, Dhumeaux D: Interferon resistance of hepatitis C virus genotype 1b:relationship to nonstructural 5A gene quasispecies mutations. J Virol 1998;72:2795–2805.
  19. Behrens SE, Tomei L, De Francesco R: Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. EMBO J 1996;15:12–22.
  20. Lohmann V, Korner F, Herian U, Bartenschlager R: Biochemical properties of hepatitis C virus NS5B RNA-dependent RNA polymerase and identification of amino acid sequence motifs essential for enzymatic activity. J Virol 1997;71:8416–8428.
  21. Desmet VJ, Gerber M, Hoofnagle JH, Manns M, Scheuer PJ: Classification of chronic hepatitis: diagnosis, grading and staging. Hepatology 1994;19:1513–1520.
  22. Kato N, Hijikata M, Ootsuyama Y, Nakagawa M, Ohkoshi S, Sugimura T, Shimotohno K: Molecular cloning of the human hepatitis C virus genome from Japanese patients with non-A, non-B hepatitis. Proc Natl Acad Sci USA 1990;87:9524–9528.
  23. Kimura M: A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980;16:111–120.
  24. Kumar S, Tamura K, Jakobsen IB, Nei M: MEGA2: Molecular Evolutionary Genetics Analysis software. Bioinformatics, 2001.
  25. Saitou N, Nei M: The neighbor-joining method: a new method for reconstruction phylogenetic trees. Mol Biol Evol 1987;4:406–425.
  26. Felsenstein J: Confidence limits on phylogenies: an approach using the bootstrap. Evolution 1985;39.783–791.

    External Resources

  27. Airaksinen A, Pariente N, Menendez-Arias L, Domingo E: Curing of foot-and-mouth disease virus from persistently infected cells by ribavirin involves enhanced mutagenesis. Virology 2003;311:339–349.
  28. Querenghi F, Yu Q, Billaud G, Maertens G, Trepo C, Zoulim F: Evolution of hepatitis C virus genome in chronically infected patients receiving ribavirin monotherapy. J Viral Hepatol 2001;8:120–131.
  29. Balzarini J, Wedgwood O, Kruining J, Pelemans H, Heijtink R, De Clercq E, McGuigan C: Anti-HIV and anti-HBV activity and resistance profile of 2′,3′-dideoxy-3′-thiacytidine (3TC) and its arylphosphoramidate derivative CF 1109. Biochem Biophys Res Commun 1996;225:363–369.
  30. Ling R, Mutimer D, Ahmed M, Boxall EH, Elias E, Dusheiko GM, Harrison TJ: Selection of mutations in the hepatitis B virus polymerase during therapy of transplant recipients with lamivudine. Hepatology 1996;24:711–713.
  31. Tipples GA, Ma MM, Fischer KP, Bain VG, Kneteman NM, Tyrrell DL: Mutation in HBV RNA-dependent DNA polymerase confers resistance to lamivudine in vivo. Hepatology 1996;24:714–717.
  32. Bartholomew MM, Jansen RW, Jeffers LJ, Reddy KR, Johnson LC, Bunzendahl H, Condreay LD, Tzakis AG, Schiff ER, Brown NA: Hepatitis-B-virus resistance to lamivudine given for recurrent infection after orthotopic liver transplantation. Lancet 1997;349:20–22.
  33. Lee JH, von Wagner M, Roth WK, Teuber G, Sarrazin C, Zeuzem S: Effect of ribavirin on virus load and quasispecies distribution in patients infected with hepatitis C virus. J Hepatol 1998;29:29–35.

 goto top of outline Author Contacts

Dr. Kazuaki Chayama
Department of Medicine and Molecular Science, Division of Frontier Medical Science
Programs for Biomedical Research, Graduate School of Biomedical Sciences
Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8551 (Japan)
Tel. +81 82 257 5190, Fax +81 82 255 6220, E-Mail chayama@hiroshima-u.ac.jp


 goto top of outline Article Information

Received: August 25, 2005
Accepted after revision: October 26, 2005
Published online: August 21, 2006
Number of Print Pages : 10
Number of Figures : 3, Number of Tables : 2, Number of References : 33


 goto top of outline Publication Details

Intervirology (International Journal of Basic and Medical Virology)

Vol. 49, No. 6, Year 2006 (Cover Date: October 2006)

Journal Editor: Liebert, U.G. (Leipzig)
ISSN: 0300–5526 (print), 1423–0100 (Online)

For additional information: http://www.karger.com/INT


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