1887

Abstract

Epstein–Barr virus (EBV) reactivation into the lytic cycle plays certain roles in the development of EBV-associated diseases, so an effective strategy to block the viral lytic cycle may be of value to reduce the disease risk or to improve the clinical outcome. This study examined whether the EBV lytic cycle could be inhibited using RNA interference (RNAi) directed against the essential viral gene Zta. In cases of EBV reactivation triggered by chemicals or by exogenous Rta, Zta-targeted RNAi prevented the induction of Zta and its downstream genes and further blocked the lytic replication of viral genomes. This antiviral effect of RNAi was not likely to be mediated by activation of the interferon pathway, as phosphorylation of STAT1 was not induced. In addition, novel EBV-infected epithelial cells showing constitutive activation of the lytic cycle were cloned; such established lytic infection was also suppressed by Zta-targeted RNAi. These results indicate that RNAi can be used to inhibit the EBV lytic cycle effectively and could also be of potential use to develop anti-EBV treatments.

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2004-06-01
2024-04-18
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References

  1. Adamson A. L., Darr D., Holley-Guthrie E., Johnson R. A., Mauser A., Swenson J., Kenney S. 2000; Epstein-Barr virus immediate-early proteins BZLF1 and BRLF1 activate the ATF2 transcription factor by increasing the levels of phosphorylated p38 and c-Jun N-terminal kinases. J Virol 74:1224–1233 [CrossRef]
    [Google Scholar]
  2. Becker J., Leser U., Marschall M., Langford A., Jilg W., Gelderblom H., Reichart P., Wolf H. 1991; Expression of proteins encoded by Epstein-Barr virus trans-activator genes depends on the differentiation of epithelial cells in oral hairy leukoplakia. Proc Natl Acad Sci U S A 88:8332–8336 [CrossRef]
    [Google Scholar]
  3. Bouvier G., Hergenhahn M., Polack A., Bornkamm G. W., de The G., Bartsch H. 1995; Characterization of macromolecular lignins as Epstein-Barr virus inducer in foodstuff associated with nasopharyngeal carcinoma risk. Carcinogenesis 16:1879–1885 [CrossRef]
    [Google Scholar]
  4. Brummelkamp T. R., Bernards R., Agami R. 2002; A system for stable expression of short interfering RNAs in mammalian cells. Science 296:550–553 [CrossRef]
    [Google Scholar]
  5. Chang Y., Sheen T. S., Lu J., Huang Y. T., Chen J. Y., Yang C. S., Tsai C. H. 1998; Detection of transcripts initiated from two viral promoters (Cp and Wp) in Epstein-Barr virus-infected nasopharyngeal carcinoma cells and biopsies. Lab Invest 78:715–726
    [Google Scholar]
  6. Chang Y., Tung C. H., Huang Y. T., Lu J., Chen J. Y., Tsai C. H. 1999; Requirement for cell-to-cell contact in Epstein-Barr virus infection of nasopharyngeal carcinoma cells and keratinocytes. J Virol 73:8857–8866
    [Google Scholar]
  7. Chang Y., Cheng S. D., Tsai C. H. 2002; Chromosomal integration of Epstein-Barr virus genomes in nasopharyngeal carcinoma cells. Head Neck 24:143–150 [CrossRef]
    [Google Scholar]
  8. Chien Y. C., Chen J. Y., Liu M. Y., Yang H. I., Hsu M. M., Chen C. J., Yang C. S. 2001; Serologic markers of Epstein-Barr virus infection and nasopharyngeal carcinoma in Taiwanese men. N Engl J Med 345:1877–1882 [CrossRef]
    [Google Scholar]
  9. Cox M. A., Leahy J., Hardwick J. M. 1990; An enhancer within the divergent promoter of Epstein-Barr virus responds synergistically to the R and Z transactivators. J Virol 64:313–321
    [Google Scholar]
  10. de-Vathaire F., Sancho-Garnier H., de-Thé H. 8 other authors 1988; Prognostic value of EBV markers in the clinical management of nasopharyngeal carcinoma (NPC): a multicenter follow-up study. Int J Cancer 42:176–181 [CrossRef]
    [Google Scholar]
  11. Dykxhoorn D. M., Novina C. D., Sharp P. A. 2003; Killing the messenger: short RNAs that silence gene expression. Nat Rev Mol Cell Biol 4:457–467 [CrossRef]
    [Google Scholar]
  12. Feederle R., Kost M., Baumann M., Janz A., Drouet E., Hammerschmidt W., Delecluse H. J. 2000; The Epstein-Barr virus lytic program is controlled by the co-operative functions of two transactivators. EMBO J 19:3080–3089 [CrossRef]
    [Google Scholar]
  13. Ge Q., McManus M. T., Nguyen T., Shen C. H., Sharp P. A., Eisen H. N., Chen J. 2003; RNA interference of influenza virus production by directly targeting mRNA for degradation and indirectly inhibiting all viral RNA transcription. Proc Natl Acad Sci U S A 100:2718–2723 [CrossRef]
    [Google Scholar]
  14. Geser A., de The G., Lenoir G., Day N. E., Williams E. H. 1982; Final case reporting from the Ugandan prospective study of the relationship between EBV and Burkitt's lymphoma. Int J Cancer 29:397–400 [CrossRef]
    [Google Scholar]
  15. Gitlin L., Andino R. 2003; Nucleic acid-based immune system: the antiviral potential of mammalian RNA silencing. J Virol 77:7159–7165 [CrossRef]
    [Google Scholar]
  16. Gitlin L., Karelsky S., Andino R. 2002; Short interfering RNA confers intracellular antiviral immunity in human cells. Nature 418:430–434 [CrossRef]
    [Google Scholar]
  17. Greenspan J. S., Greenspan D., Lennette E. T., Abrams D. I., Conant M. A., Petersen V., Freese U. K. 1985; Replication of Epstein-Barr virus within the epithelial cells of oral ‘hairy’ leukoplakia, an AIDS-associated lesion. N Engl J Med 313:1564–1571 [CrossRef]
    [Google Scholar]
  18. Grogan E., Jenson H., Countryman J., Heston L., Gradoville L., Miller G. 1987; Transfection of a rearranged viral DNA fragment, WZhet, stably converts latent Epstein-Barr viral infection to productive infection in lymphoid cells. Proc Natl Acad Sci U S A 84:1332–1336 [CrossRef]
    [Google Scholar]
  19. Henle G., Henle W. 1976; Epstein-Barr virus-specific IgA serum antibodies as an outstanding feature of nasopharyngeal carcinoma. Int J Cancer 17:1–7 [CrossRef]
    [Google Scholar]
  20. Henle G., Henle W., Clifford P. 9 other authors 1969; Antibodies to Epstein-Barr virus in Burkitt's lymphoma and control groups. J Natl Cancer Inst 43:1147–1157
    [Google Scholar]
  21. Henle W., Ho J. H., Henle G., Chau J. C., Kwan H. C. 1977; Nasopharyngeal carcinoma: significance of changes in Epstein-Barr virus-related antibody patterns following therapy. Int J Cancer 20:663–672 [CrossRef]
    [Google Scholar]
  22. Holley-Guthrie E. A., Quinlivan E. B., Mar E. C., Kenney S. 1990; The Epstein-Barr virus (EBV) BMRF1 promoter for early antigen (EA-D) is regulated by the EBV transactivators, BRLF1 and BZLF1, in a cell-specific manner. J Virol 64:3753–3759
    [Google Scholar]
  23. Jia Q., Sun R. 2003; Inhibition of gammaherpesvirus replication by RNA interference. J Virol 77:3301–3306 [CrossRef]
    [Google Scholar]
  24. Kieff E., Rickinson A. B. 2001; Epstein-Barr virus and its replication. In Fields Virology , 4th edn. pp  2511–2573 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  25. Lei K. I., Chan L. Y., Chan W. Y., Johnson P. J., Lo Y. M. 2000; Quantitative analysis of circulating cell-free Epstein-Barr virus (EBV) DNA levels in patients with EBV-associated lymphoid malignancies. Br J Haematol 111:239–246 [CrossRef]
    [Google Scholar]
  26. Levine P. H., Ablashi D. V., Berard C. W., Carbone P. P., Waggoner D. E., Malan L. 1971; Elevated antibody titers to Epstein-Barr virus in Hodgkin's disease. Cancer 27:416–421 [CrossRef]
    [Google Scholar]
  27. Liu P., Speck S. H. 2003; Synergistic autoactivation of the Epstein-Barr virus immediate-early BRLF1 promoter by Rta and Zta. Virology 310:199–206 [CrossRef]
    [Google Scholar]
  28. Lo Y. M. 2001; Quantitative analysis of Epstein-Barr virus DNA in plasma and serum: applications to tumor detection and monitoring. Ann N Y Acad Sci 945:68–72
    [Google Scholar]
  29. Lo Y. M., Chan L. Y., Lo K. W. 7 other authors 1999; Quantitative analysis of cell-free Epstein-Barr virus DNA in plasma of patients with nasopharyngeal carcinoma. Cancer Res 59:1188–1191
    [Google Scholar]
  30. Lu J., Chen S. Y., Chua H. H., Liu Y. S., Huang Y. T., Chang Y., Chen J. Y., Sheen T. S., Tsai C. H. 2000; Upregulation of tyrosine kinase TKT by the Epstein-Barr virus transactivator Zta. J Virol 74:7391–7399 [CrossRef]
    [Google Scholar]
  31. MacNeil A., Sumba O. P., Lutzke M. L., Moormann A., Rochford R. 2003; Activation of the Epstein-Barr virus lytic cycle by the latex of the plant Euphorbia tirucalli . Br J Cancer 88:1566–1569 [CrossRef]
    [Google Scholar]
  32. Manet E., Gruffat H., Trescol-Biemont M. C., Moreno N., Chambard P., Giot J. F., Sergeant A. 1989; Epstein-Barr virus bicistronic mRNAs generated by facultative splicing code for two transcriptional trans-activators. EMBO J 8:1819–1826
    [Google Scholar]
  33. Mauser A., Holley-Guthrie E., Zanation A., Yarborough W., Kaufmann W., Klingelhutz A., Seaman W. T., Kenney S. 2002; The Epstein-Barr virus immediate-early protein BZLF1 induces expression of E2F-1 and other proteins involved in cell cycle progression in primary keratinocytes and gastric carcinoma cells. J Virol 76:12543–12552 [CrossRef]
    [Google Scholar]
  34. Mellinghoff I., Daibata M., Humphreys R. E., Mulder C., Takada K., Sairenji T. 1991; Early events in Epstein-Barr virus genome expression after activation: regulation by second messengers of B cell activation. Virology 185:922–928 [CrossRef]
    [Google Scholar]
  35. Metzenberg S. 1990; Levels of Epstein-Barr virus DNA in lymphoblastoid cell lines are correlated with frequencies of spontaneous lytic growth but not with levels of expression of EBNA-1, EBNA-2, or latent membrane protein. J Virol 64:437–444
    [Google Scholar]
  36. Miller G., Lipman M. 1973; Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proc Natl Acad Sci U S A 70:190–194 [CrossRef]
    [Google Scholar]
  37. Mueller N., Evans A., Harris N. L., Comstock G. W., Jellum E., Magnus K., Orentreich N., Polk B. F., Vogelman J. 1989; Hodgkin's disease and Epstein-Barr virus. Altered antibody pattern before diagnosis. N Engl J Med 320:689–695 [CrossRef]
    [Google Scholar]
  38. Novina C. D., Murray M. F., Dykxhoorn D. M. 7 other authors 2002; siRNA-directed inhibition of HIV-1 infection. Nat Med 8:681–686
    [Google Scholar]
  39. Ragoczy T., Miller G. 1999; Role of the Epstein-Barr virus RTA protein in activation of distinct classes of viral lytic cycle genes. J Virol 73:9858–9866
    [Google Scholar]
  40. Ragoczy T., Heston L., Miller G. 1998; The Epstein-Barr virus Rta protein activates lytic cycle genes and can disrupt latency in B lymphocytes. J Virol 72:7978–7984
    [Google Scholar]
  41. Resnick L., Herbst J. S., Ablashi D. V., Atherton S., Frank B., Rosen L., Horwitz S. N. 1988; Regression of oral hairy leukoplakia after orally administered acyclovir therapy. JAMA 259:384–388 [CrossRef]
    [Google Scholar]
  42. Rickinson A. B., Kieff E. 2001; Epstein-Barr virus. In Fields Virology , 4th edn. pp  2575–2627 Edited by Knipe D. M., Howley P. M. Philadelphia: Lippincott Williams & Wilkins;
    [Google Scholar]
  43. Shao Y. M., Poirier S., Ohshima H., Malaveille C., Zeng Y., de The G., Bartsch H. 1988; Epstein-Barr virus activation in Raji cells by extracts of preserved food from high risk areas for nasopharyngeal carcinoma. Carcinogenesis 9:1455–1457 [CrossRef]
    [Google Scholar]
  44. Sledz C. A., Holko M., de Veer M. J., Silverman R. H., Williams B. R. 2003; Activation of the interferon system by short-interfering RNAs. Nat Cell Biol 5:834–839 [CrossRef]
    [Google Scholar]
  45. Stark G. R., Kerr I. M., Williams B. R., Silverman R. H., Schreiber R. D. 1998; How cells respond to interferons. Annu Rev Biochem 67:227–264 [CrossRef]
    [Google Scholar]
  46. Takase K., Kelleher C. A., Terada N., Jones J. F., Lucas J. J., Gelfand E. W. 1996; Dissociation of EBV genome replication and host cell proliferation in anti-IgG-stimulated Akata cells. Clin Immunol Immunopathol 81:168–174 [CrossRef]
    [Google Scholar]
  47. Triantos D., Porter S. R., Scully C., Teo C. G. 1997; Oral hairy leukoplakia: clinicopathologic features, pathogenesis, diagnosis, and clinical significance. Clin Infect Dis 25:1392–1396 [CrossRef]
    [Google Scholar]
  48. Tsai C. H. A., Glaser R. 1991; A comparison of Epstein-Barr virus specific proteins expressed by three Epstein-Barr virus isolates using specific monoclonal antibodies. Intervirology 32:376–382
    [Google Scholar]
  49. Tsai C. H. A., Williams M. V., Glaser R. 1991; Characterization of two monoclonal antibodies to Epstein-Barr virus diffuse early antigen which react to two different epitopes and have different biological function. J Virol Methods 33:47–52
    [Google Scholar]
  50. Tsai C. H., Liu M. T., Chen M. R., Lu J., Yang H. L., Chen J. Y., Yang C. S. 1997; Characterization of monoclonal antibodies to the Zta and DNase proteins of Epstein-Barr virus. J Biomed Sci 4:69–77 [CrossRef]
    [Google Scholar]
  51. Walling D. M., Flaitz C. M., Nichols C. M. 2003; Epstein-Barr virus replication in oral hairy leukoplakia: response, persistence, and resistance to treatment with valacyclovir. J Infect Dis 188:883–890 [CrossRef]
    [Google Scholar]
  52. Young L. S., Lau R., Rowe M. 8 other authors 1991; Differentiation-associated expression of the Epstein-Barr virus BZLF1 transactivator protein in oral hairy leukoplakia. J Virol 65:2868–2874
    [Google Scholar]
  53. Zalani S., Holley-Guthrie E., Kenney S. 1996; Epstein-Barr viral latency is disrupted by the immediate-early BRLF1 protein through a cell-specific mechanism. Proc Natl Acad Sci U S A 93:9194–9199 [CrossRef]
    [Google Scholar]
  54. Zeng Y., Zhang L. G., Wu Y. C. 7 other authors 1985; Prospective studies on nasopharyngeal carcinoma in Epstein-Barr virus IgA/VCA antibody-positive persons in Wuzhou City, China. Int J Cancer 36:545–547 [CrossRef]
    [Google Scholar]
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