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Journal of General Virology header logo Journal of General Virology

Volume 68, Issue 9

Immunoaffinity Purification of a 72K Early Antigen of Human Cytomegalovirus: Analysis of Humoral and Cell-mediated Immunity to the Purified Polypeptide No Access

Abstract

Summary

An early antigen of human cytomegalovirus (HCMV) was purified from infected cells as a 72K polypeptide by immunoaffinity chromatography using a monoclonal antibody. It was located in both nucleus and cytoplasm, and was non-glycosylated and undetectable on the surface of infected cells. Known seropositive subjects had antibody against the purified protein and it elicited proliferative T cell responses in 10 of 16 subjects. Five of 14 T cell lines established in response to the purified protein were predominantly CD8 and of these two showed major histocompatibility complex- restricted cytotoxicity against HCMV-infected cells. This provides further evidence that antigens expressed at early times may be targets for the immune response during persistent HCMV infection.

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Keyword(s): CMV and human|early antigens|T cells
© Society for General Microbiology 1987
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1987-09-01
2026-04-13

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References

  1. Bangham C. R. M., Openshaw P. J. M., Ball L. A., King A. M. Q., Wertz G. W., Askonas B. A. 1986; Human and murine cytotoxic T cells specific to respiratory syncytial virus recognise the viral nucleoprotein (N), but not the major glycoprotein (G) expressed by vaccinia virus recombinants. Journal of Immunology 137:3973–3977
     [Google Scholar]
  2. Blanton R. J., Tevethia M. J. 1981; Immunoprécipitation of virus specific immediate early and early polypeptides from cells lytically infected with human cytomegalovirus strain AD169. Virology 112:262–273
     [Google Scholar]
  3. Borysiewicz L. K., Morris S., Page J. D., Sissons J. G. P. 1983; Human cytomegalovirus specific cytotoxic T lymphocytes - requirements for in vitrogeneration and specificity. European Journal of Immunology 13:804–809
     [Google Scholar]
  4. Borysiewicz L. K., Rodgers B., Morris S., Graham S., Sisson J. G. P. 1985; Lysis of human cytomegalovirus infected fibroblasts by natural killer cells: demonstration of an IFN-independent component requiring expression of early viral proteins and characterisation of effector cells. Journal of Immunology 134:2695–2701
     [Google Scholar]
  5. Demarchi J. M. 1981; Human cytomegalovirus DNA: restriction enzyme cleavage maps and map locations for immediate-early, early and late RNAs. Virology 114:23–38
     [Google Scholar]
  6. Demarchi J. M. 1983a; Post-transcriptional control of human cytomegalovirus gene expression. Virology 124:390–402
     [Google Scholar]
  7. Demarchi J. M. 1983b; Correlation between stimulation of host cell DNA synthesis by human cytomegalovirus and lack of expression of a subset of early virus genes. Virology 129:274–286
     [Google Scholar]
  8. Fiala M., Honess R. W., Heiner D. C., Heine J. W. Jr Murnane J., Wallace R., Gube L. B. 1976; Cytomegalovirus proteins. I. Polypeptides of virions and dense bodies. Journal of Virology 19:243–254
     [Google Scholar]
  9. Forman S. J., Zaia J. A., Clark B. R., Wright C. L., Mills B. J., Pottathil R., Racklin B. C., Gallagher M. T., Wette K. 1985; A 64,000 dalton matrix protein of human cytomegalovirus induces in vitroimmune responses similar to those of whole virus antigen. Journal of Immunology 134:3391–3395
     [Google Scholar]
  10. Geballe A. P., Spaete R. R., Mocarski E. S. 1986; A as-acting element within the 5' leader on human cytomegalovirus B transcript determines kinetic class. Cell 46:858–872
     [Google Scholar]
  11. Hale A. H., Lyles D. S., Fan D. P. 1980; Elicitation of anti-Sendai virus cytotoxic T lymphocytes by viral and H2 antigens incorporated into the same lipid bilayers by membrane reconstitution into liposomes. Journal of Immunology 124:724–731
     [Google Scholar]
  12. Huang E-S. 1975; Human cytomegalovirus. III. Virus-induced DNA polymerase. Journal of Virology 16:298–310
     [Google Scholar]
  13. Kim K. S., Sapienza V. J., Carp R. I., Moon H. M. 1976; Analysis of structural polypeptides of human cytomegalovirus. Journal of Virology 24:261–276
     [Google Scholar]
  14. Lawman M. J. P., Naylor P. T., Huang L., Courtney R. J., Rouse B. T. 1981; Cell mediated immunity to herpes simplex virus : induction of cytotoxic T lymphocyte responses by viral antigens incorporated into liposomes. Journal of Immunology 126:304–308
     [Google Scholar]
  15. Mcmichael A. J., Michie C. A., Gotch F. M., Smith G. L., Moss B. 1986; Recognition of influenza A virus nucleoprotein by human cytotoxic T lymphocytes. Journal of General Virology 67:719–726
     [Google Scholar]
  16. Mocarski E. S., Pereira L., Michael N. 1985; Precise localisation 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.: 821266–1270
     [Google Scholar]
  17. Pereira L., Hoffman M., Cremer N. 1982; Electrophoretic analysis of polypeptides immunoprecipitated from cytomegalovirus infected cell extracts by human sera. Infection and Immunity 36:933–942
     [Google Scholar]
  18. Pereira L., Stagno S., Hoffman M., Volanakis J. E. 1983; Cytomegalovirus-infected cell specific polypeptides immunoprecipitated by sera from children with congenital and perinatal infections. Infection and Immunity 39:100–108
     [Google Scholar]
  19. Rasmussen L. E., Nelson R. H., Kelsall D. C., Merigan T. 1984; Murine monoclonal antibody to a single protein neutralises the infectivity of human cytomegalovirus. Proceedings of the National Academy of Sciences U.S.A.: 81876–880
     [Google Scholar]
  20. Reddehase M. J., Koszinowski U. H. 1984; Significance of herpesvirus immediate early gene expression in alveolar immunity to cytomegalovirus infection. Nature; London: 312369–371
     [Google Scholar]
  21. Reddehase M. J., Keil G. M., Kozinowski U. H. 1984; The cytolytic T lymphocyte response to murine cytomegalovirus. II. Detection of virus replication stage-specific antigens by separate populations of in vivoactive cytolytic T lymphocyte precursors. European Journal of Immunology 14:56–61
     [Google Scholar]
  22. Reddehase M. J., Bbuhring H.-J., Kozinowski U. H. 1985; Cloned long-term cytolytic T-lymphocyte line with specificity for an immediate-early membrane antigen of murine cytomegalovirus. Journal of Virology 57:408–412
     [Google Scholar]
  23. Rodgers B. C., Mundin J., Sissons J. G. P. 1985; Monoclonal antibodies recognizing early and late antigens of human cytomegalovirus : heterogeneity of polypeptides recognized between virus isolates. Journal of General Virology 66:2045–2049
     [Google Scholar]
  24. Schirm J., Roenhorst H. W., The T. H. 1980; Comparison of in vitrolymphocyte proliferation induced by cytomegalovirus-infected fibroblasts and cell free cytomegalovirus. Infection and Immunity 30:621–627
     [Google Scholar]
  25. Spaete R. R., Mocarski E. S. 1985; Regulation of cytomegalovirus gene expression: a and j3 promoters are transactivated by viral functions in permissive human fibroblasts. Journal of Virology 56:135–143
     [Google Scholar]
  26. Stenberg R. M., Witle P. R., Stinski M. F. 1985; Multiple spliced and unspliced transcripts from human cytomegalovirus immediate early region two and evidence for a common initiation site within immediate early region one. Journal of Virology 56:665–675
     [Google Scholar]
  27. Stinski M. F. 1976; Human cytomegalovirus glycoproteins associated with virion and dense bodies. Journal of Virology 19:594–609
     [Google Scholar]
  28. Stinski M. F. 1978; Sequence of protein synthesis in cells infected by human cytomegalovirus : early and late virus induced polypeptides. Journal of Virology 26:686–701
     [Google Scholar]
  29. Stinski M. F., Thomsen D. R., Stenberg R. M., Goldstein L. C. 1983; Organisation and expression of the immediate early genes of human cytomegalovirus. Journal of Virology 46:1–14
     [Google Scholar]
  30. Townsend A. R. M., Mcmichael A. J., Carter N. P., Huddleston J. A., Brownlee G. G. 1984; Cytotoxic T Cell recognition of the influenza nucleoprotein and hemagglutinin expressed in transfected mouse L cells. Cell 39:13–25
     [Google Scholar]
  31. Townsend A. R. M., Gotch F. M., Davey J. 1985; Cytotoxic T cells recognise fragments of the influenza nucleoprotein. Cell 42:457–467
     [Google Scholar]
  32. Townsend A. R. M., Rothbard J., Gotch F. M., Badhur G., Wraith D., Mcmichael A. J. 1986; The epitopes of influenza nucleoprotein recognized by cytotoxic T lymphocytes can be defined with short synthetic peptides. Cell 44:959–966
     [Google Scholar]
  33. Vaughan P. J., Banks L. M., Purifoy D. J. M., Powell K. L. 1984; Interactions between herpes simplex virus DNA-binding proteins. Journal of General Virology 65:2033–2041
     [Google Scholar]
  34. Wahren B., Robert K. H., Nordland S. 1981; Conditions for cytomegalovirus stimulation of lymphocytes. Scandinavian Journal of Immunology 13:581–586
     [Google Scholar]
  35. Wathen M. W., Stinski M. F. 1982; Temporal patterns of human cytomegalovirus transcription: mapping the viral RNAs synthesized at immediate early, early and late times after infection. Journal of Virology 41:462–477
     [Google Scholar]
  36. Wilkinson G. W. G., Akrigg A., Greenaway P. J. 1984; Transcription of the immediate early genes of human cytomegalovirus strain AD169. Virus Research 1:101–106
     [Google Scholar]
  37. Yewdell J. W., Bennink J. R., Mackett M., Lefrancois L., Lyles D. S., Moss B. 1986; Recognition of cloned vesicular stomatitis virus internal and external gene products by cytotoxic T lymphocytes. Journal of Experimental Medicine 163:1529–1538
     [Google Scholar]
  38. Zweerink H. J., Askonas B. A., Millican D., Courtneidge S. A., Skehel J. J. 1977; Cytotoxic T cells to type A influenza virus : viral haemagglutinin induces A-strain specificity while infected cells confer cross-reactive cytotoxicity. European Journal of Immunology 7:630–635
     [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-68-9-2371
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Immunoaffinity Purification of a 72K Early Antigen of Human Cytomegalovirus: Analysis of Humoral and Cell-mediated Immunity to the Purified Polypeptide
J. Gen. Virol. 68, 2371 (1987); https://doi.org/10.1099/0022-1317-68-9-2371
/content/journal/jgv/10.1099/0022-1317-68-9-2371
/content/journal/jgv/10.1099/0022-1317-68-9-2371
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