1887

Journal of General Virology header logo

Volume 72, Issue 11

Mapping of the epitopes of Epstein—Barr virus gp350 using monoclonal antibodies and recombinant proteins expressed in defines three antigenic determinants Free

Abstract

The Epstein—Barr virus (EBV) major surface membrane antigen (MA), gp350/220, induces antibodies that neutralize virus infectivity . The MA glycoprotein is encoded by nucleotides 1784 to 4504 of the HI L fragment of the EBV genome. To define the antigenic epitopes on gp350, sequences encoding portions of the protein were cloned into an expression system and eight recombinant clones were generated, two overlapping clones representing the C terminus and six overlapping clones representing the N terminus. The epitopes expressed by the recombinant proteins were mapped using 14 anti-MA monoclonal antibodies (MAbs) in a dot blot immunoassay. One of the MAbs reacted with clones that express the C terminus of gp350 and three others reacted with clones expressing the N terminal portion of the protein; the remaining MAbs tested were not reactive with the cloned proteins. The data identify three antigenic determinants on gp350. DNA sequences encoding these epitopes are located between nucleotides 1980 and 2307, 3186 and 3528, and 3528 and 3576 of the HI L fragment. In an attempt to elicit neutralizing antibodies, rabbits were immunized with gel-purified recombinant proteins from four of the clones. Neutralization assays indicate that the proteins expressed by these clones do not induce virus-neutralizing antibodies.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1991
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-11-2747
1991-11-01
2024-05-01
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/11/JV0720112747.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-11-2747&mimeType=html&fmt=ahah

References

  1. Akrand J. R., Rymo L., Walsh J. E., Bjorck E., Lindahl T., Griffin B. E. 1981; Molecular cloning of the complete Epstein-Barr virus genome as a set of overlapping restriction endonuclease fragments. Nucleic Acids Research 9:2999–3014
     [Google Scholar]
  2. Baer R., Bankier A. T., Biggin M. D., Deininger P. L., Farrell P. J., Gibson T. J., Hatfull G., Hudson G. S., Satchwell S. C., Seguin C., Tuffnell P. S., Barrell B. G. 1984; DNA sequence and expression of the B95-8 Epstein-Barr virus genome. Nature, London 310:207–211
     [Google Scholar]
  3. Beisel C., Tanner J., Matsuo T., Thorley-Lawson D., Kezdy F., Kieff E. 1985; Two major outer envelope glycoproteins of Epstein-Barr virus are encoded by the same gene. Journal of Virology 54:665–674
     [Google Scholar]
  4. Bejarano M. T., Masucci M. G., Morgan A., Morein B., Klein G., Klein E. 1990; Epstein-Barr virus (EBV) antigens processed and presented by B cells, B blasts, and macrophages trigger T-cell-mediated inhibition of EBV-induced B-cell transformation. Journal of Virology 64:139S–1401
     [Google Scholar]
  5. Biggin M., Farrell P. J., Barrell B. G. 1984; Transcription and DNA sequence of the Bam HI L fragment of B95-8 Epstein-Barr virus. EMBO Journal 3:1083–1090
     [Google Scholar]
  6. Clements G. B., Klein G., Povey S. 1975; Production by EBV infection of an EBNA-positive subline from an EBNA-negative human lymphoma cell line without detectable EBV DNA. International Journal of Cancer 16:125–133
     [Google Scholar]
  7. Edson C. M., Thorley-Lawson D. A. 1983; Synthesis and processing of the three major envelope glycoproteins of Epstein-Barr virus. Journal of Virology 46:547–556
     [Google Scholar]
  8. Emini E. A., Schleif W. A., Armstrong M. E., Silberklang M., Schultz L. D., Lehman D., Maigetter R. Z., Qualtiere L. F., Pearson G. R., Ellis R. W. 1988; Antigenic analysis of theEpstein-Barr virus major membrane antigen (gp350/220) expressed in yeast and mammalian cells: implications for the development of a subunit vaccine. Virology 166:387–393
     [Google Scholar]
  9. Epstein M. A. 1984; A prototype vaccine to prevent Epstein-Barr virus-associated tumours. Proceedings of the Royal Society B221:1–20
     [Google Scholar]
  10. Epstein M. A., Achong B. G. 1979; The 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]
  11. Epstein M. A., Achong B. G., Barr Y. M., Zajac B., Henle G., Henle W. 1966; Morphological and virological investigations on cultured Burkitt tumor lymphoblasts (strain Raji). Journal of the National Cancer Institute 37:547–559
     [Google Scholar]
  12. Epstein M. A., Morgan A. J., Finerty S., Randle B. J., Kirkwood J. K. 1985; Protection of cottontop tamarins against Epstein-Barr virus-induced malignant lymphoma by a prototype subunit vaccine. Nature, London 318:287–289
     [Google Scholar]
  13. Epstein M. A., Randle B. J., Finerty S., Kirkwood J. K. 1986; Not all potently neutralizing, vaccine-induced antibodies to Epstein-Barr virus ensure protection of susceptible experimental animals. Clinical and Experimental Immunology 63:485–490
     [Google Scholar]
  14. Fingeroth J. D., Weis J. J., Tedder T. F., Strominger J. L., Biro P. A., Fearon D. T. 1984; Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2. Proceedings of the National Academy of Sciences, U.S.A. 81:4510–4514
     [Google Scholar]
  15. Henle G., Henle W. 1966; Immunofluorescence in cells derived from Burkitt’s lymphoma. Journal of Bacteriology 91:1248–1256
     [Google Scholar]
  16. 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]
  17. Hoffman G. J., Lazarowitz S. G., Hayward S. D. 1980; Monoclonal antibody against a 250, 000-dalton glycoprotein of Epstein-Barr virus identifies a membrane antigen and a neutralizing antigen. Proceedings of the National Academy of Sciences, U.S.A. 77:2979–2983
     [Google Scholar]
  18. Hummel M., Thorley-Lawson D., Kieff E. 1984; An Epstein-Barr virus DNA fragment encodes messages for the two major envelope glycoproteins (gp350/300 and 220/200). Journal of Virology 49:413–417
     [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. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
     [Google Scholar]
  21. Lowe R. S., Keller P. M., Keech B. J., Davison A. J., Whang Y., Morgan A. J., Kieff E., Ellis R. W. 1987; Varicella-zoster virus as a live vector for the expression of foreign genes. Proceedings of the National Academy of Sciences, U.S.A. 84:3896–3900
     [Google Scholar]
  22. Mackett M., Arrand J. R. 1985; Recombinant vaccinia virus induces neutralizing antibodies in rabbits against Epstein-Barr virus membrane antigen gp340. EMBO Journal 4:3229–3234
     [Google Scholar]
  23. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
     [Google Scholar]
  24. Marcus-Sekura C. J., Woerner A. M., Klutch M., Quinnan G. V. 1988; Reactivity of an HIV gag gene polypeptide expressed in E. coli with sera from AIDS patients and monoclonal antibodies to gag. Biochimica et biophysica acta 949:213–223
     [Google Scholar]
  25. Marcus-Sekura C. J., Woerner A. M., Zhang P.-F., Klutch M. 1990a; Epitope mapping of the HIV-1 gag region by analysis of gag gene deletion fragments expressed in Escherichia coli defines eight antigenic determinants. AIDS Research and Human Retroviruses 6:317–327
     [Google Scholar]
  26. Marcus-Sekura C. J., Woerner A. M., Zweig M., Court D. L., Levin J. G., Klutch M. 1990b; Expression of HIV-1 integrase in E. coli: immunological analysis of the recombinant protein. AIDS Research and Human Retroviruses 6:1399–1408
     [Google Scholar]
  27. 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. 70:190–194
     [Google Scholar]
  28. Morgan A. J., Epstein M. A., North J. R. 1984; Comparative immunogenicity studies on Epstein-Barr virus membrane antigen (MA) gp340 with novel adjuvants in mice, rabbits, and cotton-top tamarins. Journal of Medical Virology 13:281–292
     [Google Scholar]
  29. Mullis K. B., Faloona F. A. 1987; Specific synthesis of DNA in vitro via a polymerase catalysed chain reaction. Methods in Enzymology 155:335–350
     [Google Scholar]
  30. Nemerow G. R., Wolfert R., McNaughton M. E., Cooper N. R. 1985; Identification and characterization of the Epstein-Barr virus receptor on human B lymphocytes and its relationship to the C3d complement receptor (CR2). Journal of Virology 55:347–351
     [Google Scholar]
  31. O’Farrell P. Z., Gold L. M. 1973; The identification of prereplicative bacteriophage T4 proteins. Journal of Biological Chemistry 248:5499–5501
     [Google Scholar]
  32. Qualtiere L. F., Chase R., Pearson G. R. 1982a; Purification and biologic characterization of a major Epstein-Barr virus-induced membrane glycoprotein. Journal of Immunology 129:814–818
     [Google Scholar]
  33. Qualtiere L. F., Chase R., Vorman B., Pearson G. R. 1982b; Identification of Epstein-Barr virus strain differences with monoclonal antibodies to a membrane glycoprotein. Proceedings of the National Academy of Sciences, U.S.A. 79:616–620
     [Google Scholar]
  34. Qualtiere L. F., Decoteau J. F., Nasr-El-Din M. H. 1987; Epitope mapping of the major Epstein-Barr virus outer envelope glycoprotein gp350/220. Journal of General Virology 68:535–543
     [Google Scholar]
  35. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. 1985; Enzymatic amplification of β-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 230:1350–1354
     [Google Scholar]
  36. Schultz L. D., Tanner J., Hofmann K. J., Emini E. A., Condra J. H., Jones R. E., Kieff E., Ellis R. W. 1987; Expression and secretion in yeast of a 400-kDa envelope glycoprotein derived from Epstein-Barr virus. Gene 54:113–123
     [Google Scholar]
  37. Singer C., Knauert F., Bushar G., Klutch M., Lundquist R., Quinnan G. V. 1989; Quantitation of poliovirus antigens in inactivated viral vaccines by enzyme-linked immunosorbent assay using animal sera and monoclonal antibody. Journal of Biological Standardization 17:137–150
     [Google Scholar]
  38. Sisk W. P., Chirikjian J. G., Lautenberger J., Jorcyk C., Papas T. S., Berman M. L., Zagursky R., Court D. L. 1986; A plasmid vector for cloning and expression of gene segments: expression of an HTLV-1 envelope gene segment. Gene 48:183–193
     [Google Scholar]
  39. Thorley-Lawson D. A. 1979; A virus-free immunogen effective against Epstein-Barr virus. Nature, London 281:486–488
     [Google Scholar]
  40. Thorley-Lawson D. A., Geilinger K. 1980; Monoclonal antibodies against the major glycoprotein (gp350/220) of Epstein-Barr virus neutralize infectivity. Proceedings of the National Academy of Sciences, U.S.A. 77:5307–5311
     [Google Scholar]
  41. Thorley-Lawson D. A., Poodry C. A. 1982; Identification and isolation of the main component (gp350-gp220) of Epstein-Barr virus responsible for generating neutralizing antibodies in vivo. Journal of Virology 43:730–736
     [Google Scholar]
  42. Ulaeto D., Wallace L., Morgan A., Morein B., Rickinson A. B. 1988; In vitro T cell responses to a candidate Epstein-Barr virus vaccine: human CD4+ T cell clones specific for the major envelope glycoprotein gp340. European Journal of Immunology 18:1689–1697
     [Google Scholar]
  43. Wei C.-F., Alianell G. A., Bencen G. H., Gray H. B. Jr 1983; Isolation and comparison of two molecular species of the BAL 31 nuclease from Alteromonas espejiana with distinct kinetic properties. Journal of Biological Chemistry 258:13506–13512
     [Google Scholar]
  44. Wells A., Koide N., Klein G. 1982; Two large virion envelope glycoproteins mediate Epstein-Barr virus binding to receptor-positive cells. Journal of Virology 41:286–297
     [Google Scholar]
  45. Whang Y., Silberklang M., Morgan A., Munshi S., Lenny A. B., Ellis R. W., Kieff E. 1987; Expression of the Epstein-Barr virus gp350/220 gene in rodent and primate cells. Journal of Virology 61:1796–1807
     [Google Scholar]
  46. Zweig M., Showalter S. D., Dubois G. C., Sisk W. P., Court D. L. 1987; Detection of heterologous fusion proteins in Escherichia coli with a monoclonal antibody. Gene 55:47–53
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-11-2747
Loading
Mapping of the epitopes of Epstein—Barr virus gp350 using monoclonal antibodies and recombinant proteins expressed in Escherichia coli defines three antigenic determinants
J. Gen. Virol. 72, 2747 (1991); https://doi.org/10.1099/0022-1317-72-11-2747
/content/journal/jgv/10.1099/0022-1317-72-11-2747
/content/journal/jgv/10.1099/0022-1317-72-11-2747
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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