Advertisement

In Situ Detection of Epstein-Barr Virus DNA and Viral Gene Products

  • Gerald Niedobitek
  • Hermann Herbst
Part of the Methods in Molecular Biology™ book series (MIMB, volume 174)

Abstract

Primary Epstein-Barr virus (EBV) infection is followed by a life-long persistence of the virus in the B-cell compartment of the host (1,2). Small numbers of EBV-carrying B cells have been identified in the peripheral blood as well as in lymphoid and nonlymphoid tissues of chronic virus carriers (3,4). This is relevant to the study of human tumors. The detection of EBV DNA in a tumor by polymerase chain reaction (PCR) usually does not permit conclusions as to whether this is due to the presence of the virus in the tumor cell population or to the presence of EBV-carrying “by-stander” B cells in the tissue. For a meaningful analysis of EBV infection, it is therefore necessary in many instances to establish the cellular location of the virus using morphology-based techniques.

Keywords

Cytospin Preparation Aqueous Mount Medium Nonradioactive Probe Dehydrate Section Dextran Sulphate Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Niedobitek, G. and Young, L. S. (1994) Epstein-Barr virus persistence and virus-associated tumours. Lancet 343, 333–335.PubMedCrossRefGoogle Scholar
  2. 2.
    Thorley-Lawson, D. A., Miyashita, E. M., and Khan, G. (1996) Epstein-Barr virus and the B cell: that’s all it takes. Trends Microbiol. 4, 204–208.PubMedCrossRefGoogle Scholar
  3. 3.
    Niedobitek, G., Herbst, H., Young, L. S., Brooks, L., Masucci, M. G., Crocker, J., Rickinson, A. B., and Stein, H. (1992) Patterns of Epstein-Barr virus infection in non-neoplastic lymphoid tissue. Blood 79, 2520–2526.PubMedGoogle Scholar
  4. 4.
    Hubscher, S. G., Williams, A., Davison, S. M., Young, L. S., and Niedobitek, G. (1994) Epstein-Barr virus in inflammatory diseases of the liver and liver allografts: an in situ hybridization study. Hepatology 20, 899–907.PubMedCrossRefGoogle Scholar
  5. 5.
    Niedobitek, G. and Herbst, H. (1991) Applications of in situ hybridization. Int. Rev. Exp. Pathol. 32, 1–56.PubMedGoogle Scholar
  6. 6.
    Hamilton-Dutoit, S. J. and Pallesen, G. (1994) Detection of Epstein-Barr virus small RNAs in routine paraffin sections using non-isotopic RNA/RNA in situ hybridization. Histopa-thology 25, 101–111.CrossRefGoogle Scholar
  7. 7.
    Rickinson, A. B. and Kieff, E. (1996) Epstein-Barr virus, in Fields Virology, vol. 2 (Fields, B. N., Knipe, D. M., and Howley, P. M., eds.), Lippincott-Raven, Philadelphia, pp. 2397–2446.Google Scholar
  8. 8.
    Niedobitek, G., Young, L. S., Lau, R., Brooks, L., Greenspan, D., Greenspan, J. S., and Rickinson, A. B. (1991) Epstein-Barr virus infection in oral hairy leukoplakia: virus replication in the absence of a detectable latent phase. J. Gen. Virol. 72, 3035–3046.PubMedCrossRefGoogle Scholar
  9. 9.
    Rowe, M., Lear, A., Croom-Carter, D., Davies, A. H., and Rickinson, A. B. (1992) Three pathways of Epstein-Barr virus (EBV) gene activation from EBNA1–positive latency in B lymphocytes. J. Virol. 66, 122–131.PubMedGoogle Scholar
  10. 10.
    Niedobitek, G., Herbst, H., Young, L. S., Rowe, M., Dienemann, D., Germer, C., and Stein, H. (1992) Epstein-Barr virus and carcinomas. Expression of the viral genome in an undifferentiated gastric carcinoma. Diagn. Mol. Pathol. 1, 103–108.PubMedGoogle Scholar
  11. 11.
    Herbst, H., Dallenbach, F., Hummel, M., Niedobitek, G., Pileri, S., Müller-Lantzsch, N., and Stein, H. (1991) Epstein-Barr virus latent membrane protein expression in Hodgkin and Reed-Sternberg cells. Proc. Natl. Acad. Sci. USA 88, 4766–4770.PubMedCrossRefGoogle Scholar
  12. 12.
    Grässer, F. A., Murray, P. G., Kremmer, E., Klein, K., Remberger, K., Feiden, W., et al. (1994) Monoclonal antibodies directed against the Epstein-Barr virus-encoded nuclear antigen 1 (EBNA1): immunohistologic detection of EBNA1 in the malignant cells of Hodgkin’s disease. Blood 84, 3792–3798.PubMedGoogle Scholar
  13. 13.
    Murray, P. G., Niedobitek, G., Kremmer, E., Grasser, F., Reynolds, G. M., Cruchley, A., et al. (1996) In situ detection of the Epstein-Barr virus-encoded nuclear antigen 1 in oral hairy leukoplakia and virus-associated carcinomas. J. Pathol. 178, 44–47.PubMedCrossRefGoogle Scholar
  14. 14.
    Pallesen, G., Hamilton-Dutoit, S. J., Rowe, M., and Young, L. S. (1991) Expression of Epstein-Barr virus latent gene products in tumour cells of Hodgkin’s disease. Lancet 337, 320–322.PubMedCrossRefGoogle Scholar
  15. 15.
    Miller, G. (1990) The switch between latency and replication of Epstein-Barr virus. J. Infect. Dis. 161, 833–844.PubMedGoogle Scholar
  16. 16.
    Young, L. S., Lau, R., Rowe, M., Niedobitek, G., Packham, G., Shanaham, F., et al. (1991) Differentiation-associated expression of the Epstein-Barr virus BZLF1 transactivator protein in oral “hairy” leukoplakia. J. Virol. 65, 2868–2874.PubMedGoogle Scholar
  17. 17.
    Niedobitek, G., Agathanggelou, A., Herbst, H., Whitehead, L., Wright, D. H., and Young, L. S. (1997) Epstein-Barr virus (EBV) infection in infectious mononucleosis: virus latency, replication and phenotype of EBV-infected cells. J. Pathol. 182, 151–159.PubMedCrossRefGoogle Scholar
  18. 18.
    Young, L., Alfieri, C., Hennessey, K., Evans, H., O’Hara, C., Anderson, K. C., et al. (1989) Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. N. Engl. J. Med. 321, 1080–1085.PubMedCrossRefGoogle Scholar
  19. 19.
    Rowe, M., Evans, H. S., Young, L. S., Hennessy, K., Kieff, E., and Rickinson, A. B. (1987) Monoclonal antibodies to the latent membrane protein of Epstein-Barr virus reveal heterogeneity of the protein and inducible expression in virus-transformed cells. J. Gen. Virol. 68, 1575–1586.PubMedCrossRefGoogle Scholar
  20. 20.
    Niedobitek, G., Kremmer, E., Herbst, H., Whitehead, L., Dawson, C. W., Niedobitek, E., et al. (1997) Immunohistochemical detection of the Epstein-Barr virus-encoded latent membrane protein 2A (LMP2A) in Hodgkin’s disease and infectious mononucleosis. Blood 90, 1664–1672.PubMedGoogle Scholar
  21. 21.
    Shi, S.-R., Cote, R. J., and Taylor, C. R. (1997) Antigen retrieval immunohistochemistry: past, present, and future. J. Histochem. Cytochem. 45, 327–343.PubMedGoogle Scholar
  22. 22.
    Bobrow, M. N., Harris, T. D., Shaughnessy, K. J., and Litt, G. J. (1989) Catalyzed reporter deposition, a novel method of signal amplification. J. Immunol. Methods 125, 279–285.PubMedCrossRefGoogle Scholar
  23. 23.
    Bobrow, M. N., Shaughnessy, K. J., and Litt, G. J. (1991) Catalyzed reporter deposition, a novel method of signal amplification. II. Application to membrane immunoassays. J. Immunol. Methods 137, 103–112.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2001

Authors and Affiliations

  • Gerald Niedobitek
    • 1
  • Hermann Herbst
    • 2
  1. 1.Pathologisches InstitutFriedrich-Alexander UniversitätErlangenGermany
  2. 2.Gerhard-Domagk-Institut für PathologieUniversität MünsterMünsterGermany

Personalised recommendations