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Journal of General Virology header logo Journal of General Virology

Volume 76, Issue 12

Co-expression of the Epstein—Barr virus BXLF2 and BKRF2 genes with a recombinant baculovirus produces gp85 on the cell surface with antigenic similarity to the native protein No Access

Abstract

Glycoprotein H (gH) is a conserved herpesvirus gene product functionally implicated in the penetration of the virus into the host cell. Other human herpesviruses require an accessory glycoprotein named gL for the synthesis of mature gH. We constructed a series of recombinant baculoviruses to determine whether gL expression can overcome the constraints upon Epstein—Barr virus (EBV) gH (gp85) folding and transport in this expression system. When gH and gL were co-expressed some EBV gH was transported to the insect cell surface. Deletion of 27 amino acids from the gH carboxy terminus resulted in the secretion of an 80 kDa protein (gHt) into the culture medium when it was expressed either in the presence or absence of gL and this protein could also be immunoprecipitated with E1D1. In contrast, gL was not secreted into the culture medium. Our results suggest that either co-expression of gH with gL or removal of the predicted transmembrane anchor sequence can overcome some of the constraints upon EBV gH expression in the baculovirus system.

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© Journal of General Virology 1995
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/content/journal/jgv/10.1099/0022-1317-76-12-3145
1995-12-01
2025-04-20
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References

  1. Balachandran N., Oba D. E., Hutt-Fletcher L. M. 1987; Antigenic cross-reactions among herpes simplex virus type 1 and 2, Epstein-Barr virus, and cytomegalovirus. Journal of Virology 61:1125–1135
     [Google Scholar]
  2. Cranage M. P., Smith G. L., Bell S. E., Hart H., Brown C., Bankier A. T., Tomlinson P., Barrell B. A., Brown T. C. 1988; Identification and expression of a human cytomegalovirus glycoprotein with homology to the Epstein-Barr virus BXLF2 product, varicella-zoster virus gpIII, and herpes simplex virus type I glycoprotein H. Journal of Virology 62:1416–1422
     [Google Scholar]
  3. Forghani B., Ni L. i., Grose C. 1994; Neutralization epitope of the varicella-zoster virus gH:gL glycoprotein complex. Virology 199:458–462
     [Google Scholar]
  4. Forrester A. J., Sullivan V., Simmons A., Blacklaws B. A., Smith G. L., Nash A. A., Minson A. 1991; Induction of protective immunity with antibody to herpes simplex virus type 1 glycoprotein H (gH) and analysis of the immune response to gH expressed in recombinant vaccinia virus. Journal of Virology 72:369–375
     [Google Scholar]
  5. Ghiasi H., Nesburn A. B., Wechsler S. L. 1991; Cell surface expression of herpes simplex virus type 1 glycoprotein H in recombinant baculovirus-infected cells. Virology 185:187–194
     [Google Scholar]
  6. Gompels U. A., Minson A. C. 1989; Antigenic properties and cellular localization of herpes simplex virus glycoprotein gH synthesized in a mammalian expression system. Journal of Virology 63:4744–4755
     [Google Scholar]
  7. Haddad R. S., Hutt-Fletcher L. M. 1989; Depletion of glycoprotein gp85 from virosomes made with Epstein-Barr virus proteins abolishes their ability to fuse with virus receptor-bearing cells. Journal of Virology 63:4998–5005
     [Google Scholar]
  8. Heineman T., Gong M., Sample J., Keiff E. 1988; Identification of the Epstein-Barr virus gp85 gene. Journal of Virology 62:1101–1107
     [Google Scholar]
  9. 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, USA 77:2979–2983
     [Google Scholar]
  10. Hutchinson L., Browne H., Wargent V., Davis-Poynter N., Primrac S., Goldsmith K., Minson A. C., Johnson D. C. 1992; A novel herpes simplex virus glycoprotein, gL, forms a complex with glycoprotein H (gH) and affects normal folding and surface expression of gH. Journal of Virology 66:2240–2250
     [Google Scholar]
  11. Kaye J. F., Gompels U. A., Minson A. C. 1992; Glycoprotein H of human cytomegalovirus (HCMV) forms a stable complex with the HCMV UL115 gene product. Journal of General Virology 73:2693–2698
     [Google Scholar]
  12. Li Q., Turk S. M., Hutt-Fletcher L. M. 1995; The Epstein-Barr virus (EBV) BZLF2 gene product associates with the gH and gL homologs of EBV and carries an epitope critical to infection of B cells but not epithelial cells. Journal of Virology 69:3987–3994
     [Google Scholar]
  13. Liu D. X., Gompels U. A., Foa-Tomasi L., Campadelli-Fiume G. 1993; Human herpesvirus-6 glycoprotein H and L homologs are components of the gplOO complex and the gH external domain is the target of neutralizing monoclonal antibodies. Virology 197:12–22
     [Google Scholar]
  14. McGowan E. M., Hayden M. B., Edwards S. J., Pye D., Love D. N., Whalley J. M. 1994; Expression and characterization of equine herpesvirus 1 glycoprotein H using a recombinant baculo- virus. Archives of Virology 137:389–395
     [Google Scholar]
  15. Miller N., Hutt-Fletcher L. 1988; A monoclonal antibody to glycoprotein gp85 inhibits fusion but not attachment of Epstein-Barr virus. Journal of Virology 62:2366–2372
     [Google Scholar]
  16. Morgan A. J. 1992; The development of Epstein-Barr virus vaccines. Vaccine 10:563–571
     [Google Scholar]
  17. Pulford D., Lowrey P., Morgan A. J. 1994; Expression of the Epstein-Barr virus envelope fusion glycoprotein gp85 gene by a recombinant baculovirus. Journal of General Virology 75:3241–3248
     [Google Scholar]
  18. Rickinson A. B. 1994; EBV infection and EBV-associated tumours. Symposia of the Society for General Microbiology 51:81–100
     [Google Scholar]
  19. Roberts S. R., Ponce De Leon M., Cohen G. H., Eisenberg R. J. 1991; Analysis of the intracellular maturation of the herpes simplex virus type 1 glycoprotein gH in infected and transfected cells. Virology 184:609–624
     [Google Scholar]
  20. Spaete R. R., Perot K., Scott P. I., Nelson J. A., Stinski M. F., Pachl C. 1993; Co-expression of truncated human cytomegalovirus gH with the UL115 gene product or the truncated human fibroblast growth factor receptor results in transport of gH to the cell surface. Virology 193:853–861
     [Google Scholar]
  21. Strnad B. C., Schuster T., Klein R., Hopkins R. F. III, Witmer T., Neubauer R. H., Rabin H. 1982; Production and characterization of monoclonal antibodies against Epstein-Barr virus membrane antigen. Journal of Virology 41:258–264
     [Google Scholar]
  22. Yaswen L. R., Stephens L. C., Davenport L. C., Hutt-Fletcher L. M. 1993; Epstein-Barr virus glycoprotein gp85 associates with the BKRF2 gene product and is incompletely processed as a recombinant protein. Virology 195:387–396
     [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-76-12-3145
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Co-expression of the Epstein—Barr virus BXLF2 and BKRF2 genes with a recombinant baculovirus produces gp85 on the cell surface with antigenic similarity to the native protein
J. Gen. Virol. 76, 3145 (1995); https://doi.org/10.1099/0022-1317-76-12-3145
/content/journal/jgv/10.1099/0022-1317-76-12-3145
/content/journal/jgv/10.1099/0022-1317-76-12-3145
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