1887

Abstract

Glycoprotein B (gB, gpUL55) is the major antigen recognized by the neutralizing humoral immune response against human cytomegalovirus (HCMV). The immunodominant region on gB is the antigenic domain 1 (AD-1), a complex structure that requires a minimal continuous sequence of more than 75 amino acids (aa 552–635) for antibody binding. In this study, the structural requirements for antibody binding to AD-1 have been determined. The domain was expressed in prokaryotic and eukaryotic systems and analysed in immunoblots under reducing and non-reducing conditions. In addition, AD-1 was purified in an immunologically active form and the concentration of sulphydryl groups was determined. The data clearly show that the only form that is recognized by antibodies is a disulphide-linked monomer of AD-1. The disulphide bond is formed between cysteines at amino acid positions 573 and 610 of gB.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-11-2659
2000-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/11/0812659a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-11-2659&mimeType=html&fmt=ahah

References

  1. Britt W. J., Mach M. 1996; Human cytomegalovirus glycoproteins. Intervirology 39:401–412
    [Google Scholar]
  2. Britt W. J., Vugler L. G. 1992; Oligomerization of the human cytomegalovirus major envelope glycoprotein complex gB (gp55-116). Journal of Virology 66:6747–6754
    [Google Scholar]
  3. Britt W. J., Vugler L., Butfiloski E. J., Stephens E. B. 1990; Cell surface expression of human cytomegalovirus (HCMV) gp55-116 (gB): use of HCMV-recombinant vaccinia virus-infected cells in analysis of the human neutralizing antibody response. Journal of Virology 64:1079–1085
    [Google Scholar]
  4. Earl P. L., Broder C. C., Doms R. W., Moss B. 1997; Epitope map of human immunodeficiency virus type 1 gp41 derived from 47 monoclonal antibodies produced by immunization with oligomeric envelope protein. Journal of Virology 71:2674–2684
    [Google Scholar]
  5. Eickmann M., Lange R., Ohlin M., Reschke M., Radsak K. 1998; Effect of cysteine substitutions on dimerization and interfragment linkage of human cytomegalovirus glycoprotein B (gp UL55). Archives of Virology 143:1865–1880
    [Google Scholar]
  6. Ellman G. L. 1959; A colorimetric method for determining low concentrations of mercaptans. Archives of Biochemistry and Biophysics 82:70–77
    [Google Scholar]
  7. Gonczol E., deTaisne C., Hirka G., Berencsi K., Lin W. C., Paoletti E., Plotkin S. 1991; High expression of human cytomegalovirus (HCMV)-gB protein in cells infected with a vaccinia-gB recombinant: the importance of the gB protein in HCMV immunity. Vaccine 9:631–637
    [Google Scholar]
  8. Grigera P. R., Keil W., Wagner R. R. 1992; Disulfide-bonded discontinuous epitopes on the glycoprotein of vesicular stomatitis virus (New Jersey serotype). Journal of Virology 66:3749–3757
    [Google Scholar]
  9. Kari B., Liu Y.-N. C., Goertz R., Lussenhop N., Stinski M. F., Gehrz R. 1990; Structure and composition of a family of human cytomegalovirus glycoprotein complexes designated gC-I (gB). Journal of General Virology 71:2673–2680
    [Google Scholar]
  10. Kniess N., Mach M., Fay J., Britt W. J. 1991; Distribution of linear antigenic sites on glycoprotein gp55 of human cytomegalovirus. Journal of Virology 65:138–146
    [Google Scholar]
  11. Laquerre S., Person S., Glorioso J. C. 1996; Glycoprotein B of herpes simplex virus type 1 oligomerizes through the intermolecular interaction of a 28-amino-acid domain. Journal of Virology 70:1640–1650
    [Google Scholar]
  12. Marshall G. S., Rabalais G. P., Stout G. G., Waldeyer S. L. 1992; Antibodies to recombinant-derived glycoprotein B after natural human cytomegalovirus infection correlate with neutralizing activity. Journal of Infectious Diseases 165:381–384
    [Google Scholar]
  13. Megret F., Hugnot J. P., Falconar A., Gentry M. K., Morens D. M., Murray J. M., Schlesinger J. J., Wright P. J., Young P., Van Regenmortel M. H. and others 1992; Use of recombinant fusion proteins and monoclonal antibodies to define linear and discontinuous antigenic sites on the dengue virus envelope glycoprotein. Virology 187:480–491
    [Google Scholar]
  14. Meyer H., Sundqvist V.-A., Pereira L., Mach M. 1992; Glycoprotein gp116 of human cytomegalovirus contains epitopes for strain-common and strain-specific antibodies. Journal of General Virology 73:2375–2383
    [Google Scholar]
  15. Norais N., Tang D., Kaur S., Chamberlain S. H., Masiarz F. R., Burke R. L., Marcus F. 1996; Disulfide bonds of herpes simplex virus type 2 glycoprotein gB. Journal of Virology 70:7379–7387
    [Google Scholar]
  16. Ohlin M., Sundqvist V. A., Mach M., Wahren B., Borrebaeck C. A. 1993; Fine specificity of the human immune response to the major neutralization epitopes expressed on cytomegalovirus gp58/116 (gB), as determined with human monoclonal antibodies. Journal of Virology 67:703–710
    [Google Scholar]
  17. Qadri I., Navarro D., Paz P., Pereira L. 1992; Assembly of conformation-dependent neutralizing domains on glycoprotein B of human cytomegalovirus. Journal of General Virology 73:2913–2921
    [Google Scholar]
  18. Schoppel K., Hassfurther E., Britt W. J., Ohlin M., Borrebaeck C. A., Mach M. 1996; Antibodies specific for the antigenic domain 1 (AD-1) of glycoprotein B (gpUL55) of human cytomegalovirus bind to different substructures. Virology 216:133–145
    [Google Scholar]
  19. Schoppel K., Kropff B., Schmidt C., Vornhagen R., Mach M. 1997; The humoral immune response against human cytomegalovirus is characterized by a delayed synthesis of glycoprotein-specific antibodies. Journal of Infectious Diseases 175:533–544
    [Google Scholar]
  20. Singh R., Blattler W. A., Collinson A. R. 1995; Assay for thiols based on reactivation of papain. Methods in Enzymology 251:229–237
    [Google Scholar]
  21. Speckner A., Glykofrydes D., Ohlin M., Mach M. 1999; Antigenic domain 1 of human cytomegalovirus glycoprotein B induces a multitude of different antibodies which, when combined, results in incomplete virus neutralization. Journal of General Virology 80:2183–2191
    [Google Scholar]
  22. Wagner B., Kropff B., Kalbacher H., Britt W., Sundqvist V. A., Ostberg L., Mach M. 1992; A continuous sequence of more than 70 amino acids is essential for antibody binding to the dominant antigenic site of glycoprotein gp58 of human cytomegalovirus. Journal of Virology 66:5290–5297
    [Google Scholar]
  23. Wang J. B., Adler S. P., Hempfling S., Burke R. L., Duliege A. M., Starr S. E., Plotkin S. A. 1996; Mucosal antibodies to human cytomegalovirus glycoprotein B occur following both natural infection and immunization with human cytomegalovirus vaccines. Journal of Infectious Diseases 174:387–392
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-11-2659
Loading
/content/journal/jgv/10.1099/0022-1317-81-11-2659
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