1887

Abstract

Hepatitis C virus (HCV) is a major cause of chronic hepatitis and hepatocellular carcinoma worldwide. The purpose of this study was to determine how the HCV structural proteins affect the dynamic structural and functional properties of hepatocytes and measure the extra-hepatic manifestations induced by these viral proteins. A transgenic mouse model was established by expressing core, E1 and E2 proteins downstream of a CMV promoter. HCV RNA was detected using RT-PCR in transgenic mouse model tissues, such as liver, kidney, spleen and heart. Expression of the transgene was analysed by real-time PCR to quantify viral RNA in different tissues at different ages. Immunofluorescence analysis revealed the expression of core, E1 and E2 proteins predominantly in hepatocytes. Lower levels of protein expression were detected in spleen and kidneys. HCV RNA and viral protein expression increased in the liver with age. Histological analysis of liver cells demonstrated steatosis in transgenic mice older than 3 months, which was more progressed with age. Electron microscopy analysis revealed alterations in nuclei, mitochondria and endoplasmic reticulum. HCV structural proteins induce a severe hepatopathy in the transgenic mouse model. These mice became more prone to liver and lymphoid tumour development and hepatocellular carcinoma. In this model, the extra-hepatic effects of HCV, which included swelling of renal tubular cells, were mild. It is likely that the HCV structural proteins mediate some of the histological alterations in hepatocytes by interfering with lipid transport and liver metabolism.

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2005-08-01
2019-11-20
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References

  1. Adinolfi, L. E., Gambardella, M., Andreana, A., Tripodi, M. F., Utili, R. & Ruggiero, G. ( 2001; ). Steatosis accelerates the progression of liver damage of chronic hepatitis C patients and correlates with specific HCV genotype and visceral obesity. Hepatology 33, 1358–1364.[CrossRef]
    [Google Scholar]
  2. Alter, M. J. ( 1997; ). Epidemiology of hepatitis C. Hepatology 26, 62S–65S.[CrossRef]
    [Google Scholar]
  3. Bartenschlager, R., Ahlborn-Laake, L., Mous, J. & Jacobsen, H. ( 1994; ). Kinetic and structural analyses of hepatitis C virus polyprotein processing. J Virol 68, 5045–5055.
    [Google Scholar]
  4. Bruno, S., Silini, E., Crosignani, A. & 9 other authors ( 1997; ). Hepatitis C virus genotypes and risk of hepatocellular carcinoma in cirrhosis: a prospective study. Hepatology 25, 754–758.[CrossRef]
    [Google Scholar]
  5. Bukh, J., Miller, R. H. & Purcell, R. H. ( 1995; ). Genetic heterogeneity of the hepatitis C virus. Princess Takamatsu Symp 25, 75–91.
    [Google Scholar]
  6. Chan, C. H., Hadlock, K. G., Foung, S. K. & Levy, S. ( 2001; ). V(H)1-69 gene is preferentially used by hepatitis C virus-associated B cell lymphomas and by normal B cells responding to the E2 viral antigen. Blood 97, 1023–1026.[CrossRef]
    [Google Scholar]
  7. Chiba, T., Matsuzaki, Y., Abei, M., Shoda, J., Aikawa, T., Tanaka, N. & Osuga, T. ( 1996; ). Multivariate analysis of risk factors for hepatocellular carcinoma in patients with hepatitis C virus-related liver cirrhosis. J Gastroenterol 31, 552–558.[CrossRef]
    [Google Scholar]
  8. Chisari, F. V. ( 1997; ). Cytotoxic T cells and viral hepatitis. J Clin Invest 99, 1472–1477.[CrossRef]
    [Google Scholar]
  9. Choo, Q. L., Richman, K. H., Han, J. H. & 11 other authors ( 1991; ). Genetic organization and diversity of the hepatitis C virus. Proc Natl Acad Sci U S A 88, 2451–2455.[CrossRef]
    [Google Scholar]
  10. Cohen, J. ( 1999; ). The scientific challenge of hepatitis C. Science 285, 26–30.[CrossRef]
    [Google Scholar]
  11. Davis, G. L., Esteban-Mur, R., Rustgi, V. & 8 other authors ( 1998; ). 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 339, 1493–1499.[CrossRef]
    [Google Scholar]
  12. Gasparotto, D., De, R. V. & Boiocchi, M. ( 2002; ). Hepatitis C virus, B-cell proliferation and lymphomas. Leuk Lymphoma 43, 747–751.[CrossRef]
    [Google Scholar]
  13. Grakoui, A., Wychowski, C., Lin, C., Feinstone, S. M. & Rice, C. M. ( 1993; ). Expression and identification of hepatitis C virus polyprotein cleavage products. J Virol 67, 1385–1395.
    [Google Scholar]
  14. Gretch, D. R., Polyak, S. J., Willson, R. A. & Carithers, R. L., Jr ( 1996; ). Treatment of chronic hepatitis C virus infection: a clinical and virological perspective. Adv Exp Med Biol 394, 207–224.
    [Google Scholar]
  15. Haddad, J., Deny, P., Munz-Gotheil, C., Ambrosini, J. C., Trinchet, J. C., Pateron, D., Mal, F., Callard, P. & Beaugrand, M. ( 1992; ). Lymphocytic sialadenitis of Sjogren's syndrome associated with chronic hepatitis C virus liver disease. Lancet 339, 321–323.[CrossRef]
    [Google Scholar]
  16. Hoofnagle, J. H. ( 1997; ). Hepatitis C: the clinical spectrum of disease. Hepatology 26, 15S–20S.[CrossRef]
    [Google Scholar]
  17. Hope, R. G. & McLauchlan, J. ( 2000; ). Sequence motifs required for lipid droplet association and protein stability are unique to the hepatitis C virus core protein. J Gen Virol 81, 1913–1925.
    [Google Scholar]
  18. Ikeda, K., Saitoh, S., Koida, I., Arase, Y., Tsubota, A., Chayama, K., Kumada, H. & Kawanishi, M. ( 1993; ). A multivariate analysis of risk factors for hepatocellular carcinogenesis: a prospective observation of 795 patients with viral and alcoholic cirrhosis. Hepatology 18, 47–53.[CrossRef]
    [Google Scholar]
  19. Ishikawa, T., Shibuya, K., Yasui, K., Mitamura, K. & Ueda, S. ( 2003; ). Expression of hepatitis C virus core protein associated with malignant lymphoma in transgenic mice. Comp Immunol Microbiol Infect Dis 26, 115–124.[CrossRef]
    [Google Scholar]
  20. Kato, N. ( 2000; ). Genome of human hepatitis C virus (HCV): gene organization, sequence diversity, and variation. Microb Comp Genomics 5, 129–151.[CrossRef]
    [Google Scholar]
  21. Kato, T., Miyamoto, M., Date, T. & 10 other authors ( 2003; ). Repeated hepatocyte injury promotes hepatic tumorigenesis in hepatitis C virus transgenic mice. Cancer Sci 94, 679–685.[CrossRef]
    [Google Scholar]
  22. Koike, K., Moriya, K., Ishibashi, K., Matsuura, Y., Suzuki, T., Saito, I., Iino, S., Kurokawa, K. & Miyamura, T. ( 1995; ). Expression of hepatitis C virus envelope proteins in transgenic mice. J Gen Virol 76, 3031–3038.[CrossRef]
    [Google Scholar]
  23. Koike, K., Moriya, K., Ishibashi, K., Yotsuyanagi, H., Shintani, Y., Fujie, H., Kurokawa, K., Matsuura, Y. & Miyamura, T. ( 1997; ). Sialadenitis histologically resembling Sjögren syndrome in mice transgenic for hepatitis C virus envelope genes. Proc Natl Acad Sci U S A 94, 233–236.[CrossRef]
    [Google Scholar]
  24. Kumar, D., Farrell, G. C., Fung, C. & George, J. ( 2002; ). Hepatitis C virus genotype 3 is cytopathic to hepatocytes: reversal of hepatic steatosis after sustained therapeutic response. Hepatology 36, 1266–1272.[CrossRef]
    [Google Scholar]
  25. Lemon, S. M., Lerat, H., Weinman, S. A. & Honda, M. ( 2000; ). A transgenic mouse model of steatosis and hepatocellular carcinoma associated with chronic hepatitis C virus infection in humans. Trans Am Clin Climatol Assoc 111, 146–156.
    [Google Scholar]
  26. Lerat, H., Honda, M., Beard, M. R. & 8 other authors ( 2002; ). Steatosis and liver cancer in transgenic mice expressing the structural and nonstructural proteins of hepatitis C virus. Gastroenterology 122, 352–365.[CrossRef]
    [Google Scholar]
  27. Lin, C., Lindenbach, B. D., Pragai, B. M., McCourt, D. W. & Rice, C. M. ( 1994; ). Processing in the hepatitis C virus E2-NS2 region: identification of p7 and two distinct E2-specific products with different C termini. J Virol 68, 5063–5073.
    [Google Scholar]
  28. Livak, K. J. & Schmittgen, T. D. ( 2001; ). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25, 402–408.[CrossRef]
    [Google Scholar]
  29. Lo, S. Y., Selby, M. J. & Ou, J. H. ( 1996; ). Interaction between hepatitis C virus core protein and E1 envelope protein. J Virol 70, 5177–5182.
    [Google Scholar]
  30. Mercer, D. F., Schiller, D. E., Elliott, J. F. & 9 other authors ( 2001; ). Hepatitis C virus replication in mice with chimeric human livers. Nat Med 7, 927–933.[CrossRef]
    [Google Scholar]
  31. Morales, J. M. ( 2004; ). Hepatitis C virus infection and renal disease after renal transplantation. Transplant Proc 36, 760–762.[CrossRef]
    [Google Scholar]
  32. Moriya, K., Yotsuyanagi, H., Shintani, Y., Fujie, H., Ishibashi, K., Matsuura, Y., Miyamura, T. & Koike, K. ( 1997; ). Hepatitis C virus core protein induces hepatic steatosis in transgenic mice. J Gen Virol 78, 1527–1531.
    [Google Scholar]
  33. Moriya, K., Todoroki, T., Tsutsumi, T. & 10 other authors ( 2001; ). Increase in the concentration of carbon 18 monounsaturated fatty acids in the liver with hepatitis C: analysis in transgenic mice and humans. Biochem Biophys Res Commun 281, 1207–1212.[CrossRef]
    [Google Scholar]
  34. Ohata, K., Hamasaki, K., Toriyama, K. & 11 other authors ( 2003; ). Hepatic steatosis is a risk factor for hepatocellular carcinoma in patients with chronic hepatitis C virus infection. Cancer 97, 3036–3043.[CrossRef]
    [Google Scholar]
  35. Penin, F. ( 2003; ). Structural biology of hepatitis C virus. Clin Liver Dis 7, 1–21, vii.[CrossRef]
    [Google Scholar]
  36. Perlemuter, G., Sabile, A., Letteron, P. & 8 other authors ( 2002; ). Hepatitis C virus core protein inhibits microsomal triglyceride transfer protein activity and very low density lipoprotein secretion: a model of viral-related steatosis. FASEB J 16, 185–194.[CrossRef]
    [Google Scholar]
  37. Poynard, T., Marcellin, P., Lee, S. S. & 8 other authors ( 1998; ). Randomised trial of interferon α2b plus ribavirin for 48 weeks or for 24 weeks versus interferon α2b plus placebo for 48 weeks for treatment of chronic infection with hepatitis C virus. International Hepatitis Interventional Therapy Group (IHIT). Lancet 352, 1426–1432.[CrossRef]
    [Google Scholar]
  38. Rubbia-Brandt, L., Quadri, R., Abid, K. & 8 other authors ( 2000; ). Hepatocyte steatosis is a cytopathic effect of hepatitis C virus genotype 3. J Hepatol 33, 106–115.[CrossRef]
    [Google Scholar]
  39. Sabile, A., Perlemuter, G., Bono, F. & 7 other authors ( 1999; ). Hepatitis C virus core protein binds to apolipoprotein AII and its secretion is modulated by fibrates. Hepatology 30, 1064–1076.[CrossRef]
    [Google Scholar]
  40. Santolini, E., Migliaccio, G. & La Monica, N. ( 1994; ). Biosynthesis and biochemical properties of the hepatitis C virus core protein. J Virol 68, 3631–3641.
    [Google Scholar]
  41. Shibata, M., Morizane, T., Uchida, T., Yamagami, T., Onozuka, Y., Nakano, M., Mitamura, K. & Ueno, Y. ( 1998; ). Irregular regeneration of hepatocytes and risk of hepatocellular carcinoma in chronic hepatitis and cirrhosis with hepatitis-C-virus infection. Lancet 351, 1773–1777.[CrossRef]
    [Google Scholar]
  42. Silini, E., Bottelli, R., Asti, M. & 8 other authors ( 1996; ). Hepatitis C virus genotypes and risk of hepatocellular carcinoma in cirrhosis: a case-control study. Gastroenterology 111, 199–205.[CrossRef]
    [Google Scholar]
  43. Simmonds, P., Holmes, E. C., Cha, T. A. & 7 other authors ( 1993; ). Classification of hepatitis C virus into six major genotypes and a series of subtypes by phylogenetic analysis of the NS-5 region. J Gen Virol 74, 2391–2399.[CrossRef]
    [Google Scholar]
  44. Takano, S., Yokosuka, O., Imazeki, F., Tagawa, M. & Omata, M. ( 1995; ). Incidence of hepatocellular carcinoma in chronic hepatitis B and C: a prospective study of 251 patients. Hepatology 21, 650–655.[CrossRef]
    [Google Scholar]
  45. Toussirot, E., Le Huede, G., Mougin, C., Balblanc, J. C., Bettinger, D. & Wendling, D. ( 2002; ). Presence of hepatitis C virus RNA in the salivary glands of patients with Sjogren's syndrome and hepatitis C virus infection. J Rheumatol 29, 2382–2385.
    [Google Scholar]
  46. Ueno, Y., Moriyama, M., Uchida, T. & Arakawa, Y. ( 2001; ). Irregular regeneration of hepatocytes is an important factor in the hepatocarcinogenesis of liver disease. Hepatology 33, 357–362.[CrossRef]
    [Google Scholar]
  47. Wakita, T., Taya, C., Katsume, A. & 7 other authors ( 1998; ). Efficient conditional transgene expression in hepatitis C virus cDNA transgenic mice mediated by the Cre/loxP system. J Biol Chem 273, 9001–9006.[CrossRef]
    [Google Scholar]
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