Linear antigenic and immunogenic regions of human respiratory syncytial virus N protein Free

Abstract

Three linear antigenic regions on the N protein from human respiratory syncytial virus (RSV) subgroup A (strain A2) were identified by using peptides which reacted in ELISA with sera from humans with recent or previous RSV infection. The determinants were localized within three hydrophilic regions of the protein: Thr to Gly (N3 peptide), Ser to Ala (N25 peptide) and Thr to Leu (N39 peptide). The site represented by the N39 peptide reacted with four subgroup A-specific MAbs. There were minor variations in the amino acid epitope dependencies of each of these MAbs. Two additional antigenic regions Ser to Arg and Ala to Leu, were represented by peptides that reacted with human convalescent sera, but these peptides did not differentiate between acute and convalescent sera from RSV-infected humans. Rabbit hyperimmune sera raised against selected peptides specifically precipitated different forms of the N protein from a nucleocapsid-containing homogenate derived from extracts of RSV-(subgroup A and/or B)-infected S-labelled cells in a radioimmuneprecipitation assay (RIPA); the sera were also used to demonstrate RSV infection in cells by immunofluorescent assay (IFA). Anti-N3 peptide sera precipitated N, the full-length ( 41000) form of N protein, in a RIPA done on a soluble protein pool. Anti-N39 (C-terminal region) peptide sera precipitated both forms, suggesting that N ( 38000) is an N-terminally truncated (probably at position Tyr located inside the N3 peptide linear antigenic region) form of N protein.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-76-2-357
1995-02-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/76/2/JV0760020357.html?itemId=/content/journal/jgv/10.1099/0022-1317-76-2-357&mimeType=html&fmt=ahah

References

  1. Åkerlind B., Norrby E., Örvell C., Mufson M. A. 1988; Respiratory syncytial virus: heterogeneity of subgroup B strains. Journal of General Virology 69:2145–2154
    [Google Scholar]
  2. Åkerlind-Stopner B., Utter G., Mufson M. A., Örvell C., Lerner R. A., Norrby E. 1990; A subgroup-specific antigenic site in the G protein of respiratory syncytial virus forms a disulfide-bonded loop. Journal of Virology 64:5143–5148
    [Google Scholar]
  3. Amann V. L., Lerch R. A., Anderson K., Wertz G. W. 1992; Bovine respiratory syncytial virus nucleocapsid protein: mRNA sequence analysis and expression from recombinant vaccinia virus vectors. Journal of General Virology 73:999–1003
    [Google Scholar]
  4. Banerjee A. K., Barik S. 1992; Gene expression of vesicular stomatitis virus genome RNA. Virology 188:417–428
    [Google Scholar]
  5. Barge A., Gaudin Y., Coulon P., Rujgrok R. W. H. 1993; Vesicular stomatitis virus M protein may be inside the ribonucleocapsid coil. Journal of Virology 67:7246–7253
    [Google Scholar]
  6. Barik S. 1992; Transcription of human respiratory syncytial virus genome RNA in vitro: requirement of cellular factor(s). Journal of Virology 66:6813–6818
    [Google Scholar]
  7. Bishop D. H. L., Roy P., Bean W. J., Simpson R. W. 1971; Transcription of the influenza ribonucleic acid genome by a virion polymerase. III. Completeness of the transcription process. Journal of Virology 10:689–697
    [Google Scholar]
  8. Buchholz C. J., Spehner D., Drillien R., Neubert W. J., Homann H. E. 1993; The conserved N-terminal region of Sendai virus nucleocapsid protein NP is required for nucleocapsid assembly. Journal of Virology 10:5803–5812
    [Google Scholar]
  9. Cash P., Pringle C. R., Preston C. M. 1979; The polypeptides of human respiratory syncytial virus: products of cell-free protein synthesis and post-translational modifications. Virology 92:375–384
    [Google Scholar]
  10. Collins P. L. 1991; The molecular biology of human respiratory syncytial virus (RSV) of Genus Pneumovirus. In The Paramixoviruses pp 103–162 Edited by Kingsbury D. W. New York: Plenum Press;
    [Google Scholar]
  11. Galinski M. S. 1991; Annotated nucleotide and protein sequences for selected Paramyxoviridae. In The Paramixoviruses pp 537–568 Edited by Kingsbury D. W. New York: Plenum Press;
    [Google Scholar]
  12. Garcia J., Garcia-Barreno B., Vivo A., Melero J. A. 1993; Cytoplasmic inclusions of respiratory syncytial virus-infected cells: formation of inclusion bodies in transfected cells that co-express the nucleoprotein, the phosphoprotein, and the 22K protein. Virology 195:243–247
    [Google Scholar]
  13. Gimenez H. B., Cash P., Melvin W. T. 1984; Monoclonal antibodies to human respiratory syncytial virus and their use in comparison of different virus isolates. Journal of General Virology 65:963–971
    [Google Scholar]
  14. Gombart A. F., Hirano A., Wong T. C. 1993; Conformational maturation of measles virus nucleocapsid protein. Journal of Virology 67:4133–4141
    [Google Scholar]
  15. Hill V. M., Summers D. F. 1982; Synthesis of VSV RNPs in vitro by cellular VSV RNPs added to uninfected HeLa cell extracts: VSV protein requirements for replication in vitro . Virology 123:407–419
    [Google Scholar]
  16. Houghten R. A. 1985; General method for the rapid solid phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids. Proceedings of the National Academy of Sciences, USA 82:5131–5135
    [Google Scholar]
  17. Huang Y. T., Romito R. R., De B. P., Banerjee A. K. 1993; Characterization of the in vitro system for synthesis of mRNA from human respiratory syncytial virus. Virology 193:862–867
    [Google Scholar]
  18. Joisson C., Van Regenmortel M. H. V. 1991; Influence of the C terminus of the small protein subunit of bean pod mottle virus on the antigenicity of the virus determined using monoclonal antibodies and anti-peptide antiserum. Journal of General Virology 72:2225–2232
    [Google Scholar]
  19. Leonov S. V., Utter G., Waris M., Norrby E. 1994; Linear antigenic and immunogenic regions of the respiratory syncytial virus P protein. Journal of General Virology 75:1353–1359
    [Google Scholar]
  20. McIntosh K. M., Chanock R. M. 1985; Respiratory syncytial virus. In Virology pp 1285–1304 Edited by Fields B. N., Knipe D. M. New York: Raven Press;
    [Google Scholar]
  21. Mufson M. A., Örvell C., Rafnar R. B., Norrby E. 1985; Two distinct subtypes of human respiratory syncytial virus. Journal of General Virology 66:2111–2124
    [Google Scholar]
  22. Neurath A. R., Strick N., Jiang S. 1992; Synthetic peptides and anti-peptide antibodies as probes to study interdomain interactions involved in virus assembly: the envelope of the human immuno-deficiency virus (HIV-1). Virology 188:1–13
    [Google Scholar]
  23. Norrby E., Marusyk H., Örvell C. 1970; Morphogenesis of respiratory syncytial virus in a green monkey kidney cell line (Vero). Journal of Virology 6:237–242
    [Google Scholar]
  24. Norrby E., Chen S. N., Togashi T., Sheshberadaran H., Johnson K. P. 1982; Five measles virus antigens demonstrated by use of mouse hybridoma antibodies in productively infected tissue culture cells. Archives of Virology 71:1–11
    [Google Scholar]
  25. Norrby E., Mufson M. A., Alexander H., Houghten R. A., Lerner R. A. 1987; Site-directed serology with synthetic peptides representing the large glycoprotein G of respiratory syncytial virus. Proceedings of the National Academy of Sciences, USA 84:6572–6576
    [Google Scholar]
  26. Obert G., Beyer C. 1988; An enzyme-linked immunosorbent assay using monoclonal antibodies for the detection of respiratory syncytial virus in clinical specimens. Archives of Virology 100:37–49
    [Google Scholar]
  27. Örvell C., Norrby E., Mufson M. A. 1987; Preparation and characterization of monoclonal antibodies directed against five structural components of human respiratory syncytial virus sub-group B. Journal of General Virology 68:3125–3135
    [Google Scholar]
  28. Ryan K. W., Portner A., Murti K. G. 1993; Antibodies against paramyxovirus nucleoproteins define regions important for immunogenicity and nucleocapsid assembly. Virology 183:376–384
    [Google Scholar]
  29. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  30. Ward K. A., Lambden P. R., Ogilvie M. M., Watt P. J. 1983; Antibodies to respiratory syncytial virus polypeptides and their significance in human infection. Journal of General Virology 64:1867–1876
    [Google Scholar]
  31. Waris M. 1992 Detection and antigenic variation of respiratory syncytial virus PhD thesis University of Turku; Turku, Finland:
    [Google Scholar]
  32. Yamashita T., Kawai A. 1990; Vesicular stomatitis virion-associated transcriptase activity was suppressed in vitro by a synthetic 21 amino acid oligopeptide prepared to mimic the carboxy-terminus of NS protein. Virology 178:166–173
    [Google Scholar]
  33. Zhirnov O., Bukrinskaya A. G. 1984; Nucleoprotein of animal influenza viruses, in contrast to those of human strains, are not cleaved in infected cells. Journal of General Virology 65:1127–1134
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-76-2-357
Loading
/content/journal/jgv/10.1099/0022-1317-76-2-357
Loading

Data & Media loading...

Most cited Most Cited RSS feed