1887

Abstract

Epstein-Barr virus (EBV) infects both B lymphocytes and oropharyngeal epithelium, and it has been argued that the true reservoir of virus persistence is the self-renewing basal epithelial compartment. The identification of oral hairy leukoplakia (HL) of AIDS patients as a clinically apparent focus of EBV replication in lingual epithelium therefore provides a means of studying the EBV-epithelial cell interaction . Replicative EBV DNA and productive cycle antigens are restricted to the upper, more differentiated epithelial layers in HL, and here we have applied highly sensitive hybridization and immunohistological methods to examine the lower basal/suprabasal layers for evidence of latent EBV infection. We could not detect EBV DNA in these layers using an DNA hybridization protocol which, on reference B cell lines, detected 1 viral genome/cell. Likewise, using sensitive RNA hybridization for both the small non-polyadenylated EBER RNAs (abundant transcripts seen in all known forms of EBV latency) and the latent membrane protein (LMP) mRNA (the most abundant viral mRNA in B lymphoblastoid cell lines), the basal/suprabasal cells in HL were consistently negative; immunohistological staining with specific monoclonal antibodies also gave no evidence of latently infected LMP-positive cells. When the biopsy extracts were analysed by immunoblotting with selected human antisera, in addition to abundant productive cycle antigens, a band of constant size (66K) was observed which also reacted with immunopurified antibodies monospecific for one of the latency-associated nuclear antigens, EBNA 1; the cellular origin of this EBNA 1 could not be ascertained, but it is possible that in HL the protein is expressed during the productive cycle. The absence of demonstrable EBV latency in the basal/suprabasal cells of HL suggests that this is purely a virus replicative lesion which is sustained by continual re-infection of the maturing epithelium, not by the maturation of latently infected cells from the basal compartment.

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1991-12-01
2022-08-10
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References

  1. Allday M. J., Crawford D. H. 1988; Role of epithelium in EBV persistence and pathogenesis of B-cell tumours. Lancet i:855–857
    [Google Scholar]
  2. Andersson M., Lindahl T. 1976; Epstein-Barr virus DNA in human lymphoid cell lines: in vitro conversion. Virology 73:96–105
    [Google Scholar]
  3. Busson P., Ganem G., Flores P., Mugneret F., Clausse B., Caillou B., Braham K., Wakasugi H., Lipinski M., Tursz T. 1988; Establishment and characterization of three transplantable EBV-containing nasopharyngeal carcinomas. International Journal of Cancer 42:599–606
    [Google Scholar]
  4. Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K., Stein H., Mason D. Y. 1984; Immunoenzymatic labelling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). Journal of Histochemistry and Cytochemistry 32:219–229
    [Google Scholar]
  5. Dawson C. W., Rickinson A. B., Young L. S. 1990; Epstein-Barr virus latent membrane protein inhibits human epithelial cell differentiation. Nature, London 344:777–780
    [Google Scholar]
  6. De Souza Y. G., Greenspan D., Felton J. R., Hartzog G. A., Hammer M., Greenspan J. S. 1989; Localization of Epstein-Barr virus DNA in the epithelial cells of oral hairy leukoplakia by in situ hybridization on tissue sections. New England Journal of Medicine 320:1559–1560
    [Google Scholar]
  7. Epstein M. A., Achong B. G. 1979; Relationship of the virus to Burkitt’s lymphoma. In The Epstein-Barr Virus pp 321–337 Edited by Epstein M. A., Achong B. G. Berlin, Heidelberg & New York: Springer-Verlag;
    [Google Scholar]
  8. Ernberg I., Danell E., Gratama J. W., Oosterveer M. A. P., Klein G. 1989; Molecular weight variation of EBNAs as a means to identify different Epstein-Barr virus isolates. In Epstein–Barr Virus and Human Disease pp 341–347 Edited by Ablashi D. V., Faggioni A., Kruger G. R. F., Pagano J. S., Pearson G. R. Clifton, New Jersey: The Humana Press;
    [Google Scholar]
  9. Fahraeus R., Fu H. L., Ernberg I., Finke J., Rowe M., Klein G., Falk K., Nilsson E., Yadav M., Busson P., Tursz T., Kallin B. 1988; Expression of Epstein-Barr virus-encoded proteins in nasopharyngeal carcinoma. International Journal of Cancer 42:329–338
    [Google Scholar]
  10. Fowler C. B., Reed K. D., Brannon R. B. 1989; Intranuclear inclusions correlate with the ultrastructural detection of herpes-type virions in oral hairy leukoplakia. American Journal of Surgical Pathology 13:114–119
    [Google Scholar]
  11. Gerdes J., Schwab U., Lemke H., Stein H. 1983; Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. International Journal of Cancer 31:13–20
    [Google Scholar]
  12. Gilligan K., Rajadurai P., Resnick L., Raab-Traub N. 1990; Epstein-Barr virus small nuclear RNAs are not expressed in permissively infected cells in AIDS-associated leukoplakia. Proceedings of the National Academy of Sciences, U. S. A. 87:8790–8794
    [Google Scholar]
  13. Gratama J. W., Oosterveer M. A. P., Zwaan F. E., Lepoutre J., Klein G., Ernberg I. 1988; Eradication of Epstein-Barr virus by allogeneic bone marrow transplantation: implications for the site of viral latency. Proceedings of the National Academy of Sciences, U. S. A. 85:8693–8699
    [Google Scholar]
  14. Greenspan D., Greenspan J. S., Conant M., Petersen V., Silverman S., De Souza Y. 1984; Oral ‘hairy’ leukoplakia in male homosexuals: evidence of association with both papillomavirus and a herpes-group virus. Lancet ii:831–834
    [Google Scholar]
  15. 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. New England Journal of Medicine 313:1564–1571
    [Google Scholar]
  16. Howe J. G., Steitz J. A. 1986; Localization of Epstein-Barr virus-encoded small RNAs by in situ hybridization. Proceedings of the National Academy of Sciences, U. S. A. 83:9006–9010
    [Google Scholar]
  17. Jat P., Arrand J. 1982; In vitro transcription of two Epstein-Barr virus specified small RNA molecules. Nucleic Acids Research 10:3407–3425
    [Google Scholar]
  18. Kieff E., Liebowitz D. 1990; Epstein-Barr virus and its replication. In Virology pp 1889–1920 Edited by Fields B. N., Knipe D. M. New York: Raven Press;
    [Google Scholar]
  19. Klein G. 1979; The relationship of the virus to nasopharyngeal carcinoma. In The Epstein-Barr Virus pp 339–350 Edited by Epstein M. A., Achong B. G. Berlin, Heidelberg & New York: Springer Verlag;
    [Google Scholar]
  20. Klein G., Giovanella B., Westman A., Stehlin J. S., Mumford D. 1975; An EBV-genome-negative cell line established from an American Burkitt lymphoma; receptor characteristics, EBV infectibility and permanent conversion into EBV-positive sublines by in vitro infection. Intervirology 5:319–334
    [Google Scholar]
  21. Lemon S. M., Hutt L. M., Shaw J. E., Li J. L. H., Pagano J. S. 1977; Replication of EBV in epithelial cells during infectious mononucleosis. Nature, London 268:268–270
    [Google Scholar]
  22. Lӧning T., Henke R. P., Reichart P., Becker J. 1987; In situ hybridisation to detect Epstein-Barr virus DNA in oral tissues of HIV-infected patients. Virchows Archiv A 412:127–133
    [Google Scholar]
  23. McCarrey J. R., Kaufman J. C., Churchill M. A., Zaia J. A. 1989; Differential detection of viral DNA and RNA in situ in cells infected with human cytomegalovirus. Journal of Virological Methods 25:301–314
    [Google Scholar]
  24. Melton D. A., Krieg P. A., Rebagliati M. R. 1984; Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Research 12:7035–7056
    [Google Scholar]
  25. Milani S., Herbst H., Schuppan D., Hahn E. G., Stein H. 1989; In situ hybridization for procollagen types I, III and IV mRNA in normal and fibrotic rat liver: evidence for predominant expression in nonparenchymal liver cells. Hepatology 10:84–92
    [Google Scholar]
  26. Niedobitek G., Herbst H. 1991; Applications of in situ hybridization.. International Review of Experimental Pathology 32: (in press)
    [Google Scholar]
  27. Niedobitek G., Hamilton-Dutoit S., Herbst H., Finn T., Vetner M., Pallesen G., Stein H. 1989; Identification of Epstein-Barr virus infected cells in tonsils of acute infectious mononucleosis by in situ hybridization. Human Pathology 20:796–799
    [Google Scholar]
  28. Nilsson K., Klein G., Henle W., Henle G. 1971; The establishment of lymphoblastoid cell lines from adult and from foetal human lymphoid tissue and its dependence on EBV. International Journal of Cancer 8:443–450
    [Google Scholar]
  29. Pallesen G., Hamilton-Dutoit S. J., Rowe M., Young L. S. 1991; Expression of Epstein-Barr virus latent gene products in tumor cells of Hodgkin’s disease. Lancet 337:320–322
    [Google Scholar]
  30. Polack A., Hartl G., Zimber U., Freese U. K., Laux G., Takaki K., Hohn B., Gissmann L., Bornkamm G. W. 1984; A complete set of overlapping cosmid clones of M-ABA virus derived from nasopharyngeal carcinoma and its similarity to other Epstein-Barr virus isolates. Gene 27:279–288
    [Google Scholar]
  31. Pope J. H. 1967; Establishment of cell lines from peripheral leukocytes in infectious mononucleosis. Nature, London 216:810–811
    [Google Scholar]
  32. Pringle J. H., Primrose L., Kind C. N., Talbot I. C., Lauder I. 1989; In situ hybridisation demonstration of poly-adenylated RNA sequences in formalin-fixed paraffin sections using a biotinylated oligonucleotide poly d(T) probe. Journal of Pathology 158:279–286
    [Google Scholar]
  33. Rickinson A. B., Yao Q. Y., Wallace L. E. 1985; The Epstein-Barr virus as a model of virus-host interactions. British Medical Bulletin 41:75–79
    [Google Scholar]
  34. Rowe M., Rowe D. T., Gregory C. D., Young L. S., Farrell P. J., Rupani H., Rickinson A. B. 1987a; Differences in B cell growth phenotype reflect novel patterns of Epstein-Barr virus latent gene expression in Burkitt’s lymphoma cells. EMBO Journal 6:2743–2751
    [Google Scholar]
  35. Rowe M., Evans H. S., Young L. S., Hennessy K., Kieff E., Rickinson A. B. 1987b; Monoclonal antibodies to the latent membrane protein of Epstein-Barr virus reveal heterogeneity of the protein and inducible expression in virus-transformed cells. Journal of General Virology 68:1575–1586
    [Google Scholar]
  36. Rowe M., Young L. S., Crocker J., Stokes H., Henderson S., Rickinson A. B. 1991; Epstein-Barr virus (EBV)-associated lymphoproliferative disease in the SCID mouse model: implications for the pathogenesis of EBV-positive lymphomas in man. Journal of Experimental Medicine 173:147–158
    [Google Scholar]
  37. Sample J., Kieff E. 1990; Transcription of the Epstein-Barr virus genome during latency in growth-transformed lymphocytes. Journal of Virology 64:1667–1674
    [Google Scholar]
  38. Sixbey J. W. 1989; Epstein-Barr virus and epithelial cells. In Advances in Viral Oncology vol 8 pp 187–202 Edited by Klein G. New York: Raven Press;
    [Google Scholar]
  39. Sixbey J. W., Nedrud J. G., Raab-Traub N., Hanes R. A., Pagano J. S. 1984; Epstein-Barr virus replication in oropharyngeal cells. New England Journal of Medicine 310:1225–1230
    [Google Scholar]
  40. Wang F., Petti L., Braun D., Seung S., Kieff E. 1987; A bicistronic Epstein-Barr virus mRNA encodes two nuclear proteins in latently infected, growth-transformed lymphocytes. Journal of Virology 61:945–954
    [Google Scholar]
  41. Wilson J. B., Weinberg W., Johnson R., Yuspa S., Levine A. J. 1990; Expression of the BNLF-1 oncogene of Epstein-Barr virus in the skin of transgenic mice induces hyperplasia and aberrant expression of keratin 6. Cell 61:1315–1327
    [Google Scholar]
  42. Wu T. C., Mann R. B., Charache P., Hayward S. D., Staal S., Lambe B. C., Ambinder R. F. 1990; Detection of EBV gene expression in Reed-Stemberg cells of Hodgkin’s disease. International Journal of Cancer 46:801–804
    [Google Scholar]
  43. Yao Q. Y., Ogan P., Rowe M., Wood M., Rickinson A. B. 1989a; Epstein-Barr virus-infected B cells persist in the circulation of Acyclovir-treated virus carriers. International Journal of Cancer 43:67–71
    [Google Scholar]
  44. Yao Q. Y., Ogan P., Rowe M., Wood M., Rickinson A. B. 1989b; The Epstein-Barr virus-host balance in acute infectious mononucleosis patients receiving Acyclovir anti-viral therapy. International Journal of Cancer 43:61–66
    [Google Scholar]
  45. Young L. S., Dawson C. W., Clark D., Rupani H., Busson P., Tursz T., Johnson A., Rickinson A. B. 1988; Epstein-Barr virus gene expression in nasopharyngeal carcinoma. Journal of General Virology 69:1051–1065
    [Google Scholar]
  46. Young L., Alfieri C., Hennessy K., Evans H., O’Hara C., Anderson K. C., Ritz J., Shapiro R. S., Rickinson A., Kieff E., Cohen J. I. 1989a; Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphopro-liferative disease. New England Journal of Medicine 321:1080–1085
    [Google Scholar]
  47. Young L. S., Finerty S., Brooks L., Scullion F., Rickinson A. B., Morgan A. J. 1989b; Epstein-Barr virus gene expression in malignant lymphomas induced by experimental virus infection of cottontop tamarins. Journal of Virology 63:1967–1974
    [Google Scholar]
  48. Young L. S., Lau R., Rowe M., Niedobitek G., Packham G., Shanahan F., Rowe D. T., Greenspan D., Greenspan J. S., Rickinson A. B., Farrell P. J. 1991; Differentiation-associated expression of the Epstein-Barr virus BZLF1 transactivator protein in oral hairy leukoplakia. Journal of Virology 65:2868–2874
    [Google Scholar]
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