1887

Abstract

The heterogeneity of low passage human cytomegalovirus (HCMV) strains was determined by dIII typing of 28 clinical isolates from transplant patients. These data have shown that, in general, each patient's strain has a unique restriction profile, usually comprising combinations of dIII sites present in one or more of the tissue culture-adapted strains AD169, Towne and Davis. To map sequence changes in a more refined manner we performed detailed analyses of 33 low passage clinical isolates, including those aforementioned, analysing a sequence within glycoprotein B containing a major neutralizing epitope. A 149 bp sequence containing the epitope (amino acids 608 to 625) was amplified using the polymerase chain reaction, the products were cloned and their DNA sequence was determined. Comparison of the DNA and deduced amino acid sequences with those of HCMV strain AD169 revealed that there was a high degree of conservation of the epitope between the 33 clinical isolates. However 10 of the isolates possessed silent mutations and three isolates contained mutations producing amino acid changes within the neutralizing epitope. The possible functional significance of these changes is discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-8-1985
1991-08-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/8/JV0720081985.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-8-1985&mimeType=html&fmt=ahah

References

  1. Britt W. J., Vugler L., Stephens E. B. 1988; Induction of complement-dependent and -independent neutralizing antibodies by recombinant-derived human cytomegalovirus gp55-116 (gB). Journal of Virology 62:3309–3318
    [Google Scholar]
  2. 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]
  3. Chee M. S., Bankier A. T., Beck S., Bohni R., Brown C. M., Cerny R., Horsnell T., Hutchinson C. A. III, Kouzarides T., Martignetti J. A., Preddie E., Satchwell S. C., Tomlinson P., Weston K. M., Barrell B. G. 1990; Analysis of the protein coding content of the sequence of human cytomegalovirus strain AD169. Current Topics in Microbiology and Immunology 154:125–169
    [Google Scholar]
  4. Chou S. 1989; Reactivation and recombination of multiple cytomegalovirus strains from individual organ donors. Journal of Infectious Diseases 160:11–15
    [Google Scholar]
  5. Colimon R., Somogyi T., Bertrand C., Mlchelson S. 1985; Demonstration of the colinearity of human cytomegalovirus genomes and construction of restriction maps of unknown isolates using cloned subgenomic fragments. Journal of General Virology 66:2183–2198
    [Google Scholar]
  6. Collier A. C., Chandler S. H., Handsfield H. H., Corey L., McDougall J. K. 1989; Identification of multiple strains of cytomegalovirus in homosexual men. Journal of Infectious Diseases 159:123–126
    [Google Scholar]
  7. Cranage M. P., Kouzarides T., Bankier A. T., Satchwell S., Weston K., Tomlinson P., Barrell B. G., Hart H., Bell S. E., Minson A. C., Smith G. L. 1986; Identification of the human cytomegalovirus glycoprotein B gene and induction of neutralising antibodies via its expression in recombinant vaccinia virus. EMBO Journal 5:3057–3063
    [Google Scholar]
  8. Egbert P. R., Pollar R. B., Gallagher J. G., Merigan T. C. 1980; Cytomegalovirus retinitis in immunosuppressed hosts. II. Ocular manifestations. Annals of Internal Medicine 93:664–670
    [Google Scholar]
  9. Gonczol E., Hudecz F., Ianacone J., Dietzschold B., Starr S., Plotkin S. 1986; Immune response to isolated human cytomegalovirus envelope proteins. Journal of Virology 58:661–664
    [Google Scholar]
  10. Griffiths P. D. 1990; Cytomegalovirus. In Principles and Practice of Clinical Virology pp 69–102 Edited by Zuckerman A. J., Banatvala J. E., Pattison J. R. New York: John Wiley;
    [Google Scholar]
  11. Grundy J. E., Super M., Griffiths P. D. 1986; Reinfection of a seropositive allograft recipient by cytomegalovirus from donor kidney. Lancet i:159–160
    [Google Scholar]
  12. Grundy J. E., Lui S. F., Super M., Berry N. J., Sweny P., Fernando O. N., Moorhead J., Griffiths P. D. 1988; Symptomatic cytomegalovirus infection in seropositive kidney recipients: reinfection with donor virus rather than reactivation of recipient virus. Lancet ii:132–135
    [Google Scholar]
  13. Hirt B. 1967; Selective extraction of polyoma DNA from infected mouse cell cultures. Journal of Molecular Biology 26:365–369
    [Google Scholar]
  14. Ho M. 1982 Cytomegalovirus. Biology and Infection New York & London: Plenum Press;
    [Google Scholar]
  15. Huang E. S., Alford C. A., Reynolds D. W., Stagno S., Pass R. F. 1980; Molecular epidemiology of cytomegalovirus infections in women and their infants. New England Journal of Medicine 303:958–962
    [Google Scholar]
  16. Murphy G., Kavanagh T. 1989; Speeding up the sequencing of double stranded DNA. Nucleic Acids Research 16:51–98
    [Google Scholar]
  17. Onorato I. M., Morens D. M., Martone W. J., Stansfield S. K. 1985; Epidemiology of cytomegalovirus infections: recommendations for prevention and control. Reviews of Infectious Diseases 7:479–497
    [Google Scholar]
  18. Rasmussen L., Mullenax J., Nelson R., Merigan T. 1985; Viral polypeptides detected by a complement-dependent neutralizing murine monoclonal antibody to human cytomegalovirus. Journal of Virology 55:274–280
    [Google Scholar]
  19. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Ehrlich H. A. 1988; Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  20. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd. edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  21. Utz U., Britt W., Vugler L., Mach M. 1989; Identification of a neutralizing epitope on glycoprotein gp58 of human cytomegalovirus. Journal of Virology 63:1995–2001
    [Google Scholar]
  22. Wertheim P., Buurman C., Geelen J., Van Der Noordaa J. 1983; Transmission of cytomegalovirus by renal allograft demonstrated by restriction enzyme analysis. Lancet i:980–981
    [Google Scholar]
  23. Zablotney S., Wentworth B. B., Alexander E. R. 1978; Antigenic relatedness of seventeen strains of human cytomegalovirus. American Journal of Epidemiology 107:336–343
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-8-1985
Loading
/content/journal/jgv/10.1099/0022-1317-72-8-1985
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