1887

Abstract

SUMMARY

Cytomegalovirus (CMV) encodes several glycoproteins reported to be structural homologues of glycoproteins encoded by herpes simplex virus type 1 (HSV-1). To map the antigenic and functional domains on the 907 amino acid CMV glycoprotein B (gB), we cloned and expressed a subfragment of HI fragment R of the CMV (Towne) genome into an expression vector and reacted the resulting gene product with a panel of monoclonal antibodies. Our results showed that the DNA fragment encodes related glycoproteins which we previously designated gA and which others have reported to be homologous to HSV-1 gB in CMV (AD169). Analyses of the processing of CMV gB transiently expressed in eukaryotic cells showed that glycosylation occurred independently of viral infection. Ten antibodies with complement-dependent and independent neutralizing activity reacted with a truncated derivative of gB that contained 619 amino-terminal residues but lacked the transmembrane and intracellular regions of the molecule. Twelve additional antibodies reacted with a CHO cell line expressing a 680 amino-terminal derivative of gB. All of the reactive antibodies precipitated the 447 residue carboxy-terminal cleavage product of gB from extracts of CMV-infected cells. These results showed that the neutralizing epitopes map in at least two domains of gB which are located in a discontinuous segment of 219 amino acids between residues 461 and 680 from the amino terminus of the molecule.

Keyword(s): CMV , epitope mapping and glycoprotein B
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-4-979
1989-04-01
2022-01-21
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/4/JV0700040979.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-4-979&mimeType=html&fmt=ahah

References

  1. Britt W. J. 1984; Neutralizing antibodies detect a disulfide-linked glycoprotein complex within the envelope of human cytomegalovirus. Virology 135:369–378
    [Google Scholar]
  2. Britt W. J., Auger d. 1985; Identification of a 65,000 dalton virion envelope protein of human cytomegalovirus. Virus Research 4:31–36
    [Google Scholar]
  3. Cranage M. P., Kouzarides T., Bankier A. T., Satchwell S., Weston K., Tomlinson P., Barrell B., Hart H., Bell S. E., Minson A. C., Smith G. L. 1986; Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralizing antibodies via its expression in recombinant vaccinia virus. EMBO Journal 5:3057–3063
    [Google Scholar]
  4. Cremer N., Cossen C. K., Shell G. R., Pereira L. 1985; Antibody response to cytomegalovirus polypeptides captured by monoclonal antibodies on the solid phase in enzyme immunoassays. Journal of Clinical Microbiology 21:517–521
    [Google Scholar]
  5. Graham F. L., Van Dereb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
    [Google Scholar]
  6. Gretch D. R., Kari B., Rasmussen L., Gehrz R. C., Stinski M. F. 1988; Identification and characterization of three distinct families of glycoprotein complexes in the envelopes of human cytomegalovirus. Journal of Virology 62:875–881
    [Google Scholar]
  7. Johnson D. C., Spear P. G. 1983; O-Linked oligosaccharides are acquired by herpes simplex virus glycoproteins in the Golgi apparatus. Cell 32:987–997
    [Google Scholar]
  8. Kari B., Lussenhop N., Goertz R., Wabuke-Bunoti M., Radeke R., Gehrz R. 1986; Characterization of monoclonal antibodies reactive to several biochemically distinct human cytomegalovirus glycoprotein complexes. Journal of Virology 60:345–352
    [Google Scholar]
  9. Mach M., Utz U., Fleckenstein B. 1986; Mapping of the major glycoprotein gene of human cytomegalovirus. Journal of General Virology 67:1461–1467
    [Google Scholar]
  10. Meyers I. D. 1985; Cytomegalovirus infection after organ allografting. In The Herpesviruses201 Roizman B., Lopez C. New York & London: Plenum Press;
    [Google Scholar]
  11. Middeldorp J. M., Iongsma J., The T. H. 1985; Cytomegalovirus early and late membrane antigens detected by antibodies in human convalescent sera. Journal of Virology 54:240–244
    [Google Scholar]
  12. Mocarski E. S., Pereira L., Michael N. 1985; Precise localization of genes on large animal virus genomes: use of λgtll and monoclonal antibodies to map the gene for a cytomegalovirus protein family. Proceedings of the National Academy of Sciences U.S.A 82:1266–1270
    [Google Scholar]
  13. Pereira L., Hoffman M. 1986; Immunology of human cytomegalovirus glycoproteins. Human Herpesvirus Infections: Pathogenesis, Diagnosis, and Treatment69–92 Lopez C., Roizman B. New York: Raven Press;
    [Google Scholar]
  14. Pereira L., Hoffman M., Cremer N. 1982a; Electrophoretic analysis of polypeptides immune precipitated from extracts of cytomegalovirus infected cells by human sera. Infection and Immunity 36:933–942
    [Google Scholar]
  15. Pereira L., Hoffman M., Gallo D., Cremer N. 1982b; Monoclonal antibodies to human cytomegalovirus. I.Three cell surface proteins with unique immunologic and electrophoretic properties specify cross-rective determinants. Infection and Immunity 36:924–932
    [Google Scholar]
  16. Pereira L., Stagno S., Hoffman M., Volanakis J. 1983; Cytomegalovirus infected cell polypeptides immune precipitated by children with congenital and perinatal cytomegalovirus infections. Infection and Immunity 39:100–108
    [Google Scholar]
  17. Pereira L., Hoffman M., Tatsuno M., Dondero D. 1984; aPolymorphism of human cytomegalovirus glycoproteins characterized by monoclonal antibodies. Virology 139:73–86
    [Google Scholar]
  18. Rasmussen L. E., Nelson R. M., Kelsall D. C., Merigan T. C. 1984; Murine monoclonal antibody to a single protein neutralizes the infectivity of human cytomegalovirus. Proceedings of the National Academy of Sciences U.S.A 81:876–880
    [Google Scholar]
  19. Rasmussen L., Mullenax J., Nelson M., Merigan T. C. 1985a; Human cytomegalovirus polypeptides stimulate neutralizing antibody (in vivo). Virology 145:186–190
    [Google Scholar]
  20. Rasmussen L., Mullenax J., Nelson R., Merigan T. C. 1985b; Viral polypeptides detected by a complement-dependent neutralizing murine monoclonal antibody to human cytomegalovirus. Journal of Virology 55:274–280
    [Google Scholar]
  21. Rasmussen L., Nelson M., Neff M., Merigan T. C. 1988; Characterization of two different human cytomegalovirus glycoproteins which are targets for virus neutralizing antibody. Virology 163:309–318
    [Google Scholar]
  22. Spaete R. R., Thayer R. M., Probert W. S., Masiarz F. R., Chamberlain S. H., Rasmussen L., Merigan T. C., Pachl C. 1989; Human cytomegalovirus strain Towne glycoprotein B is processed by proteolytic cleavage. Virology (in press)
    [Google Scholar]
  23. Stagno S., Pass R. F., Alford C. A. 1982; Perinatal infections: medical impact and diagnostic difficulties. Medical Virology149–181 de la Maza L., Peterson E. M. Amsterdam: Elsevier;
    [Google Scholar]
  24. Stagno S., Pass R. F., Dworsky M. E., Alford C. A. 1983; Congenital and perinatal cytomegalovirus infections. Seminars Perinatal 7:31–42
    [Google Scholar]
  25. Uchida N., Smilowitz M., Tanzer M. L. 1979; Monovalent ionophores inhibit secretion of procollagen and fibronectin from cultured human fibroblasts. Proceedings of the National Academy of Sciences U.S.A 76:1868–1972
    [Google Scholar]
  26. Wong G. G., Witek J. S., Temple P. A., Wilkens K. M., Leary A. C., Luxenberg D. P., Jones S. S., Brown E. L., Kay R. M., Orr E. C., Shoemaker C., Golde D. W., Kaufman R. J., Hewick R. M., Wang E. A., Clark S. C. 1985; Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins. Science 228:810–812
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-70-4-979
Loading
/content/journal/jgv/10.1099/0022-1317-70-4-979
Loading

Data & Media loading...

Most cited this month 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