1887

Abstract

Monoclonal antibody (MAb) E-13 to human cytomegalovirus is used widely for diagnostic and fundamental studies, and has been shown to be directed against an immediate early (IE) protein(s). To determine which viral antigen is detected by MAb E-13, four subfragments from the open reading frame encoded by exons 2, 3 or 4 of IE-1 were cloned in the bacterial expression vector pROS. The resulting fusion proteins contained amino acids 77 to 491 encoded by mainly exon 4, amino acids 25 to 78 encoded by exon 3, amino acids 1 to 85 encoded by exons 2 and 3, and amino acids 1 to 24 encoded by exon 2. The reactivity of MAb E-13 with the fusion proteins was assayed by Western blotting. MAb E-13 was shown to react exclusively with proteins encoded by exon 2 and therefore recognizes IE proteins which contain the N-terminal amino acid sequence encoded by exon 2, namely the major 72K IE protein, the 82K to 86K IE-2 protein and the 52K to 55K IE-2 protein. MAb E-13 can be used to detect both IE-1- and IE-2-encoded proteins, which share the polypeptide encoded by exon 2.

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1992-10-01
2022-01-20
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References

  1. Akrigg A., Wilkinson G. W., Oram J. D. 1985; The structure of the major immediate early gene of human cytomegalovirus strain AD169. Virus Research 2:107–121
    [Google Scholar]
  2. Alpert G., Mazeron M. C., Colimon R., Plotkin S. 1985; Rapid detection of human cytomegalovirus in the urine of humans. Journal of Infectious Diseases 152:631–633
    [Google Scholar]
  3. Boland G. J., De Gast G. C., Hené R. J., Jambroes G., Donckerwolcke R., The T. H., Mudde G. C. 1990; Early detection of active cytomegalovirus (CMV) infection after heart and kidney transplantation by testing for immediate early antigenemia and influence of cellular immunity on the occurence of CMV infection. Journal of Clinical Microbiology 28:2069–2075
    [Google Scholar]
  4. Boshart M., Weber F., Jahn G., Dorsch-Hasler K., Flecken-stein B., Schaffner W. 1985; A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell 41:521–530
    [Google Scholar]
  5. Chang C. P., Malone C. L., Stinski M. F. 1989; A human cytomegalovirus early gene has three inducible promoters that are regulated differentially at various times after infection. Journal of Virology 63:281–290
    [Google Scholar]
  6. Chee M., Rudolph S. A., Plachter B., Barrell B., Jahn G. 1989; Identification of the major capsid protein gene of human cytomegalovirus. Journal of Virology 63:1345–1353
    [Google Scholar]
  7. Cherrington J. M., Khoury E. L., Mocarski E. S. 1991; Human cytomegalovirus IE2 negatively regulates α gene expression via a short target sequence near the transcription start site. Journal of Virology 65:887–896
    [Google Scholar]
  8. Depto A. S., Stenberg R. M. 1989; Regulated expression of the human cytomegalovirus pp65 gene: octamer sequence in the promoter is required for activation by viral gene products. Journal of Virology 63:1232–1238
    [Google Scholar]
  9. Ellinger S., Glockshuber R., Jahn G., Plückthun A. 1989; Cleavage of procaryotically expressed human immunodeficiency virus fusion proteins by factor Xa and application in Western blot (immunoblot) assays. Journal of Clinical Microbiology 27:971–976
    [Google Scholar]
  10. Goldstein L. C., McDougall J. M., Hackman R., Meyers J. D., Donnal Thomas E., Nowinski R. C. 1982; Monoclonal antibodies to cytomegalovirus: rapid identification of clinical isolates and preliminary use in diagnosis of cytomegalovirus pneumonia. Infection and Immunity 38:273–281
    [Google Scholar]
  11. Gӧnczol E., Andrews P. W., Plotkin S. A. 1984; Cytomegalovirus replicates in differentiated but not in undifferentiated human embryonal carcinoma cells. Science 224:159–161
    [Google Scholar]
  12. Hermiston T. W., Malona C. L., Witte P. R., Stinski M. F. 1987; Identification and characterization of human cytomegalovirus immediate early region-2 gene that stimulates gene expression from an inducible promoter. Journal of Virology 61:3214–3221
    [Google Scholar]
  13. Kadonaga J. T., Kristin R., Masiarz F. R., Than R. 1987; Isolation of cDNA encoding transcription factor SP1 and functional analysis of the DNA binding domain. Cell 51:1079–1090
    [Google Scholar]
  14. LaFemina R. L., Hayward G. S. 1986; Constitutive and retinoic acid-inducible expression of cytomegalovirus immediate-early genes in human teratocarcinoma cells. Journal of Virology 58:434–440
    [Google Scholar]
  15. LaFemina R. L., Hayward G. S. 1988; Differences in cell type-specific blocks to immediate-early gene expression and DNA replication of human, simian and murine cytomegalovirus. Journal of General Virology 69:355–374
    [Google Scholar]
  16. LaFemina R. L., Pizzorno M. C., Mosca J. D., Hayward G. S. 1989; Expression of the acidic nuclear immediate-early protein (IE1) of human cytomegalovirus in stable cell lines and its preferential association with metaphase chromosomes. Virology 172:584–600
    [Google Scholar]
  17. Lautenschlager I., Suni J., Ahonen J., Grӧnhagen-Riska C., Ruutu P., Ruutu T., Tukieainen P. 1989; Detection of cytomegalovirus by the early-antigen immunofluorescence test versus conventional tissue culture. European Journal of Clinical Microbiology and Infectious Diseases 8:610–613
    [Google Scholar]
  18. Liu B., Hermiston T. W., Stinski M. F. 1991; A cis-acting element in the major immediate-early (IE) promoter of human cytomegalovirus is required for negative regulation by IE-2. Journal of Virology 65:897–903
    [Google Scholar]
  19. Mackenzie D., McLaren L. C. 1989; Increased sensitivity for rapid detection of cytomegalovirus by shell vial centrifugation assay using mink lung cell cultures. Journal of Virological Methods 26:183–188
    [Google Scholar]
  20. Malone C. L., Vesole D. H., Stinski M. F. 1990; Transactivation of a human cytomegalovirus early promoter by gene products from the immediate-early gene IE-2 and augmentation by IE-1: mutational analysis of the viral proteins. Journal of Virology 64:1498–1506
    [Google Scholar]
  21. Mazeron M. C., Berbar T., Guillemin M. C., Colimon R., Roseto A., Pérol Y. 1983; Production d’anticorps mono-clonaux contre le cytomégalovirus humain. Compte Rendu de VAcademie des Sciences, Paris 297:305–308
    [Google Scholar]
  22. Mazeron M. C., Colimon R., Roseto A., Perol Y. 1984; Detection of cytomegaloviremia using monoclonal antibodies. Developments in Biological Standardization 57:287–291
    [Google Scholar]
  23. Otto S. M., Sullivan-Tailyour G., Malone C. L., Stinski M. F. 1988; Subcellular localization of the major immediate-early protein (IE-1) of human cytomegalovirus at early times after infection. Virology 61:190–199
    [Google Scholar]
  24. Pizzorno M. C., Hayward G. S. 1990; The IE-2 gene products of human cytomegalovirus specifically down-regulate expression from the major immediate-early promoter through a target sequence located near the cap site. Journal of Virology 64:6154–6165
    [Google Scholar]
  25. Said G., Lacroix C., Chemouilli P., Goulon-Goeau C., Roullet E., Penaud E., De Broucker T., Meduri G., Vincent D., Vittcoq D., Vildé J. L. 1991; Cytomegalovirus neuropathy in acquired immunodeficiency syndrome: a clinical and pathological study. Annals of Neurology 29:139–146
    [Google Scholar]
  26. Spatz A., Tricottet V., Saliba F., Samuel D., Bismuth H., Reynѐs M. 1990; Diagnostic rapide des hépatites à cytomégalovirus apres transplantation hépatique. Annales de Pathologic 10:331–335
    [Google Scholar]
  27. Stenberg R. M., Stinski M. F. 1985; Autoregulation of the human cytomegalovirus major immediate-early gene. Journal of Virology 56:676–682
    [Google Scholar]
  28. Stenberg R. M., Thomsen D. R., Stinski M. F. 1984; Structural analysis of the major immediate-early gene of human cytomegalovirus. Journal of Virology 49:190–199
    [Google Scholar]
  29. Stenberg R. M., Depto A. S., Fortney J., Nelson J. A. 1989; Regulated expression of early and late RNAs and proteins from the human cytomegalovirus immediate-early gene region. Journal of Virology 63:2699–2708
    [Google Scholar]
  30. Stenberg R. M., Fortney J., Barlow S. W., Magram B. P., Nelson J. A., Ghazal P. 1990; Promoter-specific trans activation and repression by human cytomegalovirus immediate-early proteins involve common and unique domains. Journal of Virology 64:1556–1565
    [Google Scholar]
  31. Stinski M. F., Thomsen D. R., Stenberg R. M., Goldstein L. C. 1983; Organization and expression of the immediate early genes of human cytomegalovirus. Journal of Virology 46:1–14
    [Google Scholar]
  32. Thomsen D. R., Stenberg R. M., Goins W. F., Stinski M. F. 1984; Promoter-regulatory region of the major immediate early gene of human cytomegalovirus. Proceedings of the National Academy of Sciences, U.S.A. 81:659–663
    [Google Scholar]
  33. Torkey C. B., Carrigan D. R. 1989; Immunohistochemical detection of an immediate early antigen of human cytomegalovirus in normal tissues. Journal of Infectious Diseases 160:741–751
    [Google Scholar]
  34. Van Der Loos C. M., Volkers H. H., Rook R., Van Den Berg F. M., Houthoff H. J. 1989; Simultaneous application of in situ DNA hybridization and immunochemistry on one tissue section. Histochemical Journal 21:279–284
    [Google Scholar]
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