1887

Abstract

The attachment protein, G, of human respiratory syncytial virus (RSV) is an 84K to 90K species which has a high content of -linked and -linked carbohydrates. The unglycosylated form of this protein was expressed by inserting a full-length cDNA copy of the mRNA from the A2 strain of RSV into a prokaryotic expression vector under the control of the lambda P promoter. cells transformed with the G-containing plasmid synthesized a protein of 40000 that specifically reacted with polyclonal and two neutralizing monoclonal antibodies raised against the native RSV G glycoprotein. Recombinant G protein was purified by immunoaffinity chromatography using a neutralizing monoclonal antibody. Cotton rats immunized with the recombinant G protein produced serum antibodies to the G glycoprotein that neutralized RSV . The study demonstrates that the G protein of RSV can be expressed in bacteria and that at least one neutralizing epitope is not structurally dependent on carbohydrates.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-74-3-453
1993-03-01
2021-10-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/74/3/JV0740030453.html?itemId=/content/journal/jgv/10.1099/0022-1317-74-3-453&mimeType=html&fmt=ahah

References

  1. Anderson L. J., Hierholzer J. C., Tsou C, Hendry R. M., Fkrnie B. F., Stone Y., McIntosh K. 1985; Antigenic characterization of respiratory syncytial virus strains with monoclonal antibodies. Journal of Infectious Diseases 151:626–633
    [Google Scholar]
  2. Bialy H. 1987; Recombinant proteins virtual authenticity. Bio/Technology 5:883–890
    [Google Scholar]
  3. Bullas L. R., Ryu J. I. 1983; Salmonella typhimurium LT2 strains which are r-m+ for all three chromosomally located systems of DNA restriction and modification. Journal of Bacteriology 156:471–474
    [Google Scholar]
  4. Chanock R. M., Kim H. W., Brandt C. D., Parrot R. H. 1982; Respiratory syncytial virus. In Viral Infections in Humans: Epidemiology and Control pp. 471–489 Edited by Evans A. S. New York: Plenum Press;
    [Google Scholar]
  5. Drahos D., Szybalski W. 1981; Antitermination and termination functions of the cloned nut L, N and t LI modules of coliphage lambda. Gene 16:261–274
    [Google Scholar]
  6. Elango N., Prince G. A., Murphy B. R., Venkatesan S., Chanock R. M., Moss B. 1986; Resistance to human respiratory syncytial virus (RSV) infection induced by immunization of cotton rats with recombinant vaccinia virus expressing the RSV glycoprotein. Proceedings of the National Academy of Sciences, U.S.A. 83:1906–1910
    [Google Scholar]
  7. Elus R. W., Gerety R. 1990; Key issues in the selection of an expression system for vaccine antigens. Journal of Medical Virology 31:54–58
    [Google Scholar]
  8. Garcia-Barreno B., Palomo C., Penas C., Delgado T., Perez-Brena P., Melero J. A. 1989; Marked differences in the antigenic structure of human respiratory syncytial virus F and G glycoproteins. Journal of Virology 63:925–932
    [Google Scholar]
  9. Gottesman M. E., Adhya S., Das A. 1981; Transcription antitermination by bacteriophage lambda N gene product. Journal of Molecular Biology 140:57–75
    [Google Scholar]
  10. Huang Y. T., Collins P. L., Wertz G. W. 1985; Characterization of ten proteins of human respiratory syncytial virus: identification of a fourth envelope-associated protein. Virus Research 2:157–173
    [Google Scholar]
  11. Johnson P. R., Olmsted R. A., Prince G. A., Murphy B. R., Alling D. W., Walsh E. E., Collins P. L. 1987; Antigenic relatedness between glycoproteins of human respiratory syncytial virus subgroups A and B : evaluation of the contributions of F and G glycoproteins to immunity. Journal of Virology 61:3163–3166
    [Google Scholar]
  12. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  13. Leshinskaya N. P., Pokrovska E. E., Kantorovitch E. A., Grigorieva S. K., Shivartsman S. Ya. 1988; Ontogenesis of the formation of secretory antibodies to respiratory syncytial (RS) virus. Epidemiology and Infection 101:565–575
    [Google Scholar]
  14. Levine S., Klaiber-Franco R., Paradiso P. R. 1987; Demonstration that glycoprotein G is the attachment protein of respiratory syncytial virus. Journal of General Virology 68:2521–2524
    [Google Scholar]
  15. McIntosh K., Chanock R. M. 1990; Respiratory syncytial virus. In Virology, 2nd edn. pp. 1045–1072 Edited by Fields B. N., Knipe D. M. New York: Raven Press;
    [Google Scholar]
  16. Mufson M. A., Örvell C, Rafnar B., Norrby E. 1985; Two distinct subtypes of human respiratory syncytial virus. Journal of General Virology 66:2111–2124
    [Google Scholar]
  17. Olmsted R. A., Elango N., Prince G. A., Murphy B. R., Johnson P. R., Moss B., Chanock R. M., Collins P. L. 1986; Expression of the F glycoprotein of respiratory syncytial virus by a recombinant vaccinia virus: comparison of the individual contributions of the F and G glycoproteins to host immunity. Proceedings of the National Academy of Sciences, U.S.A. 83:7462–7466
    [Google Scholar]
  18. Palomo C., García-Barreno B., Peñas C., Melero J. A. 1991; The G protein of human respiratory syncytial virus : significance of carbohydrate side-chains and the C-terminal end to its antigenicity. Journal of General Virology 72:669–675
    [Google Scholar]
  19. Prince G. A., Horswood R. L., Chanock R. M. 1985; Quantitative aspects of passive immunity to respiratory syncytial virus infection in infant cotton rats. Journal of Virology 57:517–520
    [Google Scholar]
  20. Reed R. R. 1981; Transposon-mediated site-specific recombination: a defined in vitro system. Cell 25:713–719
    [Google Scholar]
  21. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A. 74:5463–5467
    [Google Scholar]
  22. Satake M., Coligan J. E., Elango N., Norrby E., Venkatesan S. 1985; Respiratory syncytial virus envelope glycoprotein (G) has a novel structure. Nucleic Acids Research 13:7795–7812
    [Google Scholar]
  23. Towbin H., Staehelin T., Gordon J. 1979; Electrophoretic transfer of proteins from polysacrylamide gels to nitrocellulose sheets : procedure and some applications. Proceedings of the National Academy of Sciences, U.S.A. 76:4350–4354
    [Google Scholar]
  24. Wagner D. K., Muelenaer P., Henderson F. W., Snyder M. H., Reimer C. B., Walsh E. E., Anderson L. J., Nelson D. L., Murphy B. R. 1989; Serum immunoglobulin G antibody subclass response to respiratory syncytial virus F and G glycoprotein after first, second and third infections. Journal of Clinical Microbiology 27:589–592
    [Google Scholar]
  25. Walsh E. E., Hruska J. 1983; Monoclonal antibodies to respiratory syncytial virus proteins. Identification of the fusion protein. Journal of Virology 41:171–177
    [Google Scholar]
  26. Walsh E. E., Schlesinger J. J., Brandriss M. W. 1984; Purification and characterization of GP90, one of the envelope glycoproteins of respiratory syncytial virus. Journal of General Virology 65:761–767
    [Google Scholar]
  27. Walsh E. E., Brandriss M. W., Schlesinger J. J. 1987; Immunological differences between the envelope glycoproteins of two strains of human respiratory syncytial virus. Journal of General Virology 68:2169–2176
    [Google Scholar]
  28. Walsh E. E., Hall C. B., Schlesinger J. J., Brandriss M. W., Hildreth S., Paradiso P. 1989; Comparison of antigenic sites of subtype-specific respiratory syncytial virus attachment proteins. Journal of General Virology 70:2953–2961
    [Google Scholar]
  29. Wertz G. W., Collins P. L., Huang Y., Gruber C., Levine S., Ball L. A. 1985; Nucleotide sequence of the G protein gene of human respiratory syncytial virus reveals an unusual type of viral membrane protein. Proceedings of the National Academy of Sciences, U.S.A. 82:4075–4084
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-74-3-453
Loading
/content/journal/jgv/10.1099/0022-1317-74-3-453
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