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.

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2000-11-01
2019-12-13
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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.[CrossRef]
    [Google Scholar]
  6. Ellman, G. L. ( 1959; ). A colorimetric method for determining low concentrations of mercaptans. Archives of Biochemistry and Biophysics 82, 70-77.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [Google Scholar]
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