1887

Abstract

Summary

Monoclonal antibodies specific for the ‘latent membrane protein’ (LMP) of Epstein-Barr virus (EBV), one of the effector proteins of EBV-induced B cell transformation, have been generated from mice immunized with a -galactosidase fusion protein containing the carboxyl half of the B95. 8 strain LMP sequence. Four monoclonal IgG antibodies, designated CS. 1, CS. 2, CS. 3 and CS. 4, which together recognized at least three different epitopes on the molecule, were used to examine various aspects of LMP expression in B cell lines transformed . The pooled CS. 1 to 4 reagent detected the LMPs encoded by each of 20 geographically distinct EBV isolates, despite a degree of inter-isolate heterogeneity in the size and antigenicity of the protein. In cell lines carrying the prototype B95. 8 virus strain, particularly if these were virus producers, an additional lower molecular weight LMP was also detected; this appeared to correspond to the truncated form of the protein already predicted to exist from the analysis of B95. 8 lytic cycle mRNAs. Attempts were made to identify an analogous truncated form of LMP in cell lines carrying other virus isolates after treatment with phorbol ester and/or sodium butyrate to induce virus production. Surprisingly these experiments showed that expression of the full length LMP molecule was itself strongly inducible by these agents; when monitored at the single cell level, this was a generalized response and was not restricted to cells entering a lytic cycle. Expression of LMP in EBV-transformed B cells therefore appears to be subject to a distinct type of regulation.

Keyword(s): EBV , latent membrane protein and MAbs
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1987-06-01
2024-04-19
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References

  1. Burnette W. N. 1981; “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated Protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  2. Dambaugh T., Hennessy K., Fennewald S., Kieff E. 1986; The EBV genome and its expression in latent infection. In The Epstein-Barr Virus: Recent Advances pp. 14–45 Epstein M. A., Achong B. G. Edited by London: Heinemann;
    [Google Scholar]
  3. De Thé G. 1982; Epidemiology of Epstein-Barr virus and associated diseases in man. In The Herpesviruses 1 pp. 25–103 Roizman B. Edited by New York & London: Plenum Press;
    [Google Scholar]
  4. Fennewald S., Van Santen V, Kieff E. 1984; Nucleotide sequence of an mRNA transcribed in latent growth-transforming virus infection indicates that it may encode a membrane protein. Journal of Virology 51:411–419
    [Google Scholar]
  5. Griffith I. p. 1972; Immediate visualization of proteins in dodecyl sulfate-polyacrylamide gels by prestaining with remazole dyes. Analytical Biochemistry 46:402–412
    [Google Scholar]
  6. Hennessy K., Fennewald S., Hummel M., Cole T., Kieff E. 1984; A membrane protein encoded by Epstein-Barr virus in latent growth transforming infection. Proceedings of the National Academy of Sciences U.S.A: 817207–7211
    [Google Scholar]
  7. Hudson G. S., Farrell P. J., Barrell B. G. 1985; Two related but differentially expressed potential membrane proteins encoded by the EcoRI Dhet region of Epstein-Barr virus B95-8. Journal of Virology 53:528–535
    [Google Scholar]
  8. Johnson G. D., Davidson R. S., McNamee K. C., Russell G., Goodwin D., Holborrow E. J. 1982; Fading of immunofluorescence during microscopy: a study of this phenomenon and its remedy. Journal of Immunological Methods 55:231–242
    [Google Scholar]
  9. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature; London: 227680–685
    [Google Scholar]
  10. Liebowitz D., Wang D., Kieff E. 1986; Orientation and patching of the latent infection membrane protein encoded by Epstein-Barr virus. Journal of Virology 58:233–237
    [Google Scholar]
  11. Mann K. P., Staunton D., Thorley-Lawson D. A. 1985; Epstein-Barr virus-encoded protein found in plasma membranes of transformed cells. Journal of Virology 55:710–720
    [Google Scholar]
  12. Miller G., Lipman M. 1973; Release of infectious Epstein-Barr virus by transformed marmoset leukocytes. Proceedings of the National Academy of Sciences U.S.A: 70190–194
    [Google Scholar]
  13. Miller J. F. 1972; Assay of β-galactosidase. In Experiments in Molecular Genetics pp. 352–355 Miller J. F. Edited by New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  14. Rickinson A. B. 1986; Cellular immunological responses to the virus infection. In The Epstein-Barr Virus: Recent Advances pp. 75–125 Epstein M. A., Achong B. G. Edited by London: Heinemann;
    [Google Scholar]
  15. Rickinson A. B., Young L. S., Rowe M. 1987; Influence of the Epstein-Barr virus nuclear antigen EBNA 2 on the growth phenotype of virus transformed B cells. Journal of Virology in press
    [Google Scholar]
  16. Rooney C. M., Gregory C. D., Rowe M., Finerty S., Edwards C., Rupani H., Rickinson A. B. 1986; Endemic Burkitt’s lymphoma: phenotypic analysis of Burkitt’s lymphoma biopsy cells and of the derived tumour cell lines. Journal of the National Cancer Institute 77:681–687
    [Google Scholar]
  17. Rowe D. T., Rowe M., Evan G. I., Wallace L. E., Farrell P. J., Rickinson A. B. 1986; Restricted expression of EBV latent genes and T-lymphocyte-detected membrane antigen in Burkitt’s lymphoma cells. EMBO Journal 5:2599–2607
    [Google Scholar]
  18. Rowe M., Hildreth J. E. K., Rickinson A. B., Epstein M. A. 1982; Monoclonal antibodies to Epstein-Barr virus-induced, transformation-associated cell surface antigens: binding patterns and effect upon virus-specific T-cell cytotoxicity. International Journal of Cancer 29:373–381
    [Google Scholar]
  19. Rowe M., Rooney C. M., Rickinson A. B., Lenoir G. M., Rupani H., Moss D. J., Stein H., Epstein M. A. 1985; Distinctions between endemic and sporadic forms of Epstein-Barr virus-positive Burkitt’s lymphoma. International Journal of Cancer 35:435–441
    [Google Scholar]
  20. Rüther U., Müller-Hill B. 1983; Easy identification of cDNA clones. EMBO Journal 2:1791–1794
    [Google Scholar]
  21. Skare J., Farley J., Strominger J. L., Fresen K. O., Cho M. S., Zur Hausen H. 1985; Transformation by Epstein-Barr virus requires DNA sequences in the region of BamHI fragments Y and H. Journal of Virology 55:286–297
    [Google Scholar]
  22. Van Santen V., Cheung A., Kieff E. 1981; Epstein-Barr virus RNA. VII. Size and direction of transcription of virus-specified cytoplasmic RNAs in a transformed cell line. Proceedings of the National Academy of Sciences U.S.A: 781930–1934
    [Google Scholar]
  23. Wang D., Liebowitz D., Kieff E. 1985; An EBV membrane protein expressed in immortalized lymphocytes transforms established rodent cells. Cell 43:831–840
    [Google Scholar]
  24. Wang F., Gregory C. D., Rowe M., Rickinson A. B., Wang D., Birkenbach M., Kikutani H., Kishimoto T., Kieff E. 1987; Epstein-Barr virus nuclear protein 2 specifically induces expression of the B cell activation antigen, CD23. Proceedings of the National Academy of Sciences U.S.A: in press
    [Google Scholar]
  25. Yates J. L., Warren N., Sugden B. 1985; Stable replication of plasmids derived from Epstein-Barr virus in various mammalian cells. Nature; London: 313812–815
    [Google Scholar]
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