1887

Abstract

Summary

We have isolated reactive clones from a λgt11 expression library of human cytomegalovirus (HCMV) DNA using HCMV-positive human sera. Among the recombinant clones obtained, one carried a fragment encoding a portion of p52, the major non-structural DNA-binding protein of 52K (p52) and another carried a part of the gene coding for p150, the major structural phosphoprotein. These two fusion proteins were examined by immunoblot analysis to test their ability to bind specific antibodies in human sera. The results showed that high titres of antibody to the DNA-binding protein are present in sera of patients undergoing acute HCMV infection, whereas high titres of antibodies to the structural phosphoprotein are widespread in the healthy HCMV-seropositive population. The use of these fusion proteins as antigens for differential screening of serum as a way of detecting recent HCMV infection is discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-5-1247
1989-05-01
2021-10-21
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/5/JV0700051247.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-5-1247&mimeType=html&fmt=ahah

References

  1. Braun D. J., Pereira L., Norrild B., Roizman B. 1983; Application of denatured, electrophoretically separated and immobilized lysates of herpes simplex virus infected cells for detection of monoclonal antibodies and for the study of the properties of viral proteins. Journal of Virology 46:103–112
    [Google Scholar]
  2. Burnette W. N. 1981; “Western blotting”: electrophoretic transfer of proteins from sodium dodecyl sulfate–polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Analytical Biochemistry 112:195–203
    [Google Scholar]
  3. Geballe A. P., Leach F. S., Mocarski E. S. 1986; Regulation of cytomegalovirus late gene expression. Alpha genes are controlled by post-transcriptional events. Journal of Virology 57:864–874
    [Google Scholar]
  4. Gibson W. 1983; Protein counterparts of human and simian cytomegaloviruses. Virology 128:391–406
    [Google Scholar]
  5. Ho M. 1982 Cytomegalovirus: Biology and Infection119–204 Greenough W. R. III, Merigan T. New York: Plenum Press;
    [Google Scholar]
  6. Jahn G., Scholl B.-C., Traupe B., Fleckenstein B. 1987; The two major phosphoproteins (pp65 and pp150) of human cytomegalovirus and their antigenic properties. Journal of General Virology 68:1327–1337
    [Google Scholar]
  7. Landini M. P., Ripalti A. 1982; A DNA nicking activity associated with nucleocapsid of human cytomegalovirus. Archives of Virology 73:351–356
    [Google Scholar]
  8. Landini M. P., Re M. C., Mirolo G., Baldassarri B., La Placa M. 1985; Human immune response to cytomegalovirus structural polypeptides studied by immunoblotting. Journal of Medical Virology 17:303–311
    [Google Scholar]
  9. Landini M. P., Mirolo G., Cooppolecchia P., Re M. C., La Placa M. 1986; Serum antibodies to individual cytomegalovirus structural polypeptides in renal transplant recipients during viral infection. Microbiology and Immunology 30:683–695
    [Google Scholar]
  10. Meyer H., Bankier T., Landini M. P., Brown C. M., Barrell B. G., Rüger R., Mach M. 1988; Identification and procaryotic expression of the gene coding for the highly immunogenic 28 kilodalton structural phosphoprotein (pp28). Journal of Virology 62:2243–2250
    [Google Scholar]
  11. Mocarski E. S., Pereira L., Michael N. 1985; Precise localization of genes on large animal genomes: use of λgt11 and monoclonal antibodies to map the gene for a cytomegalovirus protein family. Proceedings of the National Academy of SciencesU.S.A 82:1266–1270
    [Google Scholar]
  12. Pereira L., Hoffman M., Tatsuno M., Dondero D. 1984; Polymorphism of human cytomegalovirus glycoproteins characterized by monoclonal antibodies. Virology 139:73–86
    [Google Scholar]
  13. Quinnan G. V., Masur H., Rook A. H., Armstrong G., Frederick W. R., Epstein J., Manischewitz M. S., Macher M. D., Jackson L., Ames J., Smith H. A., Parker M., Pearson G. R., Parillo J., Mitchell C., Straus S. E. 1984; Herpesvirus infection in the acquired immunodeficiency syndrome. Journal of the American Medical Association 252:72–77
    [Google Scholar]
  14. Ripalti A., Landini M. P., La Placa M. 1988; A 46 Kd polypeptide, present in purified cytomegalovirus is provided with DNase activity and is antigenically related to a higher molecular weight, enzymatically inactive cellular protein. Microbiologica 11:131–140
    [Google Scholar]
  15. Scholl B.-C., Von Hintzenstern J., Borisch B., Traupe B., Bröker M., Jahn G. 1988; Prokaryotic expression of immunogenic polypeptides of the large phosphoprotein (ppl50) of human cytomegalovirus. Journal of General Virology 69:1195–1204
    [Google Scholar]
  16. Smith D. E., Fisher P. A. 1984; Identification, developmental regulation and response to heat shock of two antigenically related forms of a major nuclear envelope protein in Drosophila embryos: application of an improved method for affinity purification of antibodies using polypeptides immobilized on nitrocellulose blots. Journal of Cell Biology 99:20–28
    [Google Scholar]
  17. Thomsen D. R., Stinski M. F. 1981; Cloning of the human cytomegalovirus genome as XbaI endonuclease fragments. Gene 16:207–216
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-70-5-1247
Loading
/content/journal/jgv/10.1099/0022-1317-70-5-1247
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