1887

Abstract

SUMMARY

Preparative SDS-polyacrylamide gel electrophoresis has been used in the purification of the gp340 component of Epstein-Barr (EB) virus-determined membrane antigen (MA), in tractable quantities, from the B95-8 marmoset lymphoblastoid cell line. Successful renaturation of the purified molecule was achieved. This procedure gave a 50-fold increase in the recovery of antigen compared to conventional techniques. The data suggest that the antigenic sites recognized by human sera containing antibodies to MA are largely confined to the protein portion of the molecule. An eightfold improvement in the yield of gp340 was obtained when B95-8 cells were cultured in the presence of 12--tetradecanoyl-phorbol-13-acetate. Gel filtration studies indicate that the major polypeptide components of MA are not associated in detergent solution. Immunization of rabbits with purified and renatured gp340 resulted in the generation of high-titre antisera which were specific for gp340, demonstrating that antigen prepared by this procedure is suitable for further evaluation as an experimental vaccine against EB virus infection.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-64-2-455
1983-02-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/jgv/64/2/JV0640020455.html?itemId=/content/journal/jgv/10.1099/0022-1317-64-2-455&mimeType=html&fmt=ahah

References

  1. Bowen B., Steinberg J., Laemmli U. K., Weintraub H. 1980; The detection of DNA binding proteins by protein blotting. Nucleic Acids Research 8:1–20
    [Google Scholar]
  2. Crumpton M. J., Snary D. 1974; Preparation and properties of lymphocyte plasma membrane. Contemporary Topics in Molecular Immunology 3:27–56
    [Google Scholar]
  3. De Schryver A., Klein G., Hewetson J., Rocchi G., Henle W., Henle G., Moss D. J., Pope J. H. 1974; Comparison of EBV neutralization tests based on abortive infection or transformation of lymphoid cells and their relation to membrane reactive antibodies (anti-MA). International Journal of Cancer 13:353–362
    [Google Scholar]
  4. Edson C. M., Thorley-Lawson D. A. 1981; Epstein Barr virus membrane antigens: characterisation, distribution, and strain differences. Journal of Virology 39:172–184
    [Google Scholar]
  5. Epstein M. A. 1976; Epstein–Barr virus - is it time to develop a vaccine program?. Journal of the National Cancer Institute 56:697–700
    [Google Scholar]
  6. Epstein M. A., Achong B. G. (editors) 1979; The relationship of the virus to Burkitt’s lymphoma. In The Epstein–Barr Virus pp 321–337 Berlin, Heidelberg & New York: Springer-Verlag;
    [Google Scholar]
  7. Epstein M. A., North J. R. 1982; Characterisation of Epstein–Barr virus antigens: towards a vaccine for malignancies associated with the virus. In Monoclonal Antibodies in Clinical Medicine pp 277–300 Edited by McMichael A. J., Fabre J. W. London: Academic Press;
    [Google Scholar]
  8. Franklin S. M., North J. R., Morgan A. J., Epstein M. A. 1981; Antigenic differences between the membrane antigen polypeptides determined by different EB virus isolates. Journal of General Virology 53:371–376
    [Google Scholar]
  9. Gergely L., Klein G., Ernberg I. 1971; Appearance of Epstein–Barr virus-associated antigens in infected Raji cells. Virology 45:19–21
    [Google Scholar]
  10. Hager D. A., Burgess R. R. 1980; Elution of proteins from sodium dodecyl sulphate-polyacrylamide gels, removal of sodium dodecyl sulphate, and renaturation of enzymatic activity: Results with sigma subunit of Escherichia coli RNA polymerase, wheat germ DNA topiosomerase, and other enzymes. Analytical Biochemistry 109:76–86
    [Google Scholar]
  11. Henle G., Henle W. 1979; The virus as the etiologic agent of infectious mononucleosis. In The Epstein–Barr Virus pp 297–320 Edited by Epstein M. A., Achong B. G. Berlin, Heidelberg & New York: Springer-Verlag;
    [Google Scholar]
  12. Hoffman G. F., Lazarowitz S. G., Hayward S. D. 1980; Monoclonal antibody against a 250000-dalton glycoprotein of Epstein–Barr virus identifies a membrane antigen and a neutralising antigen. Proceedings of the National Academy of Sciences of the United States of America 77:2979–2983
    [Google Scholar]
  13. Hudewentz J., Bornkamm G. W., Zur Hausen H. 1980; Effect of the diterpene ester TPA on Epstein–Barr virus antigen and DNA synthesis in producer and non-producer cell lines. Virology 100:175–178
    [Google Scholar]
  14. 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]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  16. Miller G., Shope T., Lisco H., Stitt D., Lipman M. 1972; Epstein–Barr virus: transformation, cytopathic changes, and viral antigens in squirrel monkey and marmoset leukocytes. Proceedings of the National Academy of Sciences of the United States of America 69:383–387
    [Google Scholar]
  17. Mueller-Lantzsch N., Georg-Fries B., Herbst H., Zur-Hausen H., Braun D. G. 1981; Epstein–Barr virus strain and group specific antigenic determinants detected by monoclonal antibodies. International Journal of Cancer 28:321–327
    [Google Scholar]
  18. North J. R., Morgan A. J., Epstein M. A. 1980; Observations on the EB virus envelope and virus-determined membrane antigen (MA) polypeptides. International Journal of Cancer 26:231–240
    [Google Scholar]
  19. North J. R., Morgan A. J., Thompson J. L., Epstein M. A. 1982a; Quantification of an EB virus-associated membrane antigen (MA) component. Journal of Virological Methods 5:55–65
    [Google Scholar]
  20. North J. R., Morgan A. J., Thompson J. L., Epstein M. A. 1982b; Purified EB virus gp340 induces potent virus-neutralising antibodies when incorporated in liposomes. Proceedings of the National Academy of Sciences of the United States of America (in press)
    [Google Scholar]
  21. Pearson G., Dewey F., Klein G., Henle G., Henle W. 1970; Relation between neutralization of Epstein–Barr virus and antibodies to cell-membrane antigens induced by the virus. Journal of the National Cancer Institute 45:989–995
    [Google Scholar]
  22. Pearson G., Henle G., Henle W. 1971; Production of antigens associated with Epstein–Barr virus in experimentally infected lymphoblastoid cell lines. Journal of the National Cancer Institute 46:1243–1250
    [Google Scholar]
  23. Qualtiere L. F., Pearson G. R. 1980; Radioimmune precipitation study comparing the Epstein–Barr virus membrane antigens expressed on P, HR-1 virus-superinfected Raji cells to those expressed on cells in a B95-8 virus-transformed producer culture activated with tumor-promoting agent (TPA). Virology 102:360–369
    [Google Scholar]
  24. Strnad B. C., Schuster T., Klein R., Hopkins R. F. III, Witmer T., Neubauer R. H., Rabin H. 1982; Production and characterisation of monoclonal antibodies against the Epstein–Barr virus membrane antigen. Journal of Virology 41:258–264
    [Google Scholar]
  25. Thorley-Lawson D. A. 1979; A virus-free immunogen effective against Epstein–Barr virus. Nature, London 281:486–488
    [Google Scholar]
  26. Thorley-Lawson D. A., Edson C. M. 1979; The polypeptides of the Epstein–Barr virus membrane antigen complex. Journal of Virology 32:458–467
    [Google Scholar]
  27. Thorley-Lawson D. A., Geilinger K. 1980; Monoclonal antibodies against the major glycoprotein (gp 350/220) of Epstein–Barr virus neutralise infectivity. Proceedings of the National Academy of Sciences of the United States of America 77:5307–5311
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-64-2-455
Loading
/content/journal/jgv/10.1099/0022-1317-64-2-455
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error