1887

Abstract

SUMMARY

A baculovirus expression vector (AcLeon) derived from nuclear polyhedrosis virus (AcNPV) was prepared containing the complete 6·6 kb coding region of the P3/Leon/37 strain of poliovirus type 3 placed under the control of the AcNPV polyhedrin promoter. The recombinant virus was used to infect insect cells. As demonstrated by use of the appropriate antibodies, infected insect cells made poliovirus proteins that included the structural proteins VP0, VP1 and VP3. Poliovirus particles were recovered from extracts of the infected cells and demonstrated to be free from detectable levels of RNA and to be non-infectious in tissue culture. After particle purification by CsC1 gradient centrifugation and immunization of outbred mice, antibodies to the structural proteins, including neutralizing antibodies, were obtained. Other recombinant baculoviruses, containing the majority of the capsid coding region of P3/Leon/37 (e.g. AcCAP21, nucleotide residues 742 to 3318), made an unprocessed precursor to the poliovirus structural proteins. These data suggested that processing of the poliovirus gene product by the AcLeon construct was catalysed by the poliovirus-encoded proteases. The results demonstrated that antigenic and immunogenic poliovirus proteins and empty particles can be made in insect cells by recombinant baculoviruses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-70-6-1453
1989-06-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/70/6/JV0700061453.html?itemId=/content/journal/jgv/10.1099/0022-1317-70-6-1453&mimeType=html&fmt=ahah

References

  1. Blondel B., Crainic R., Fichot O., Dufraisse G., Cardrea A., Girard M., Horaud F. 1986; Mutations conferring resistance to neutralization with monoclonal antibodies in type 1 poliovirus can be located outside or inside the antibody binding site. Journal of Virology 57:81–90
    [Google Scholar]
  2. Brown M., Faulkner P. 1977; A plaque assay for nuclear polyhedrosis viruses using a solid overlay. Journal of General Virology 36:361–364
    [Google Scholar]
  3. Burke K. L., Dunn G., Ferguson M., Minor P. D., Almond J. W. 1988; Antigen chimaeras of poliovirus as potential new vaccines. Nature, London 322:81–82
    [Google Scholar]
  4. Clarke B. E., Sangar D. V. 1988; Processing and assembly of foot-and-mouth disease virus proteins using subgenomic RNA. Journal of General Virology 69:2313–2325
    [Google Scholar]
  5. Diamond D. C., Jameson B. A., Brown J., Kohara M., Abe S., Ithoh H., Komatsu T., Arita M., Kuge S., Osterhaus A. D. M. E., Crainic R., Nomoto A., Wimmer E. 1985; Antigenic variation and resistance to neutralization in poliovirus type 1. Science 229:1090–1093
    [Google Scholar]
  6. Hanecak R., Semler B. L., Anderson C. W., Wimmer E. 1982; Proteolytic processing of poliovirus polypeptides: antibodies to polypeptide P3-7c inhibit cleavage at glutamine-glycine pairs. Proceedings of the National Academy of Sciences, U.S.A 79:3973–3977
    [Google Scholar]
  7. Hogle J. M., Chow M., Filman D. I. 1985; The three-dimensional structure of poliovirus at 2·9 Å resolution. Science 229:1358–1365
    [Google Scholar]
  8. Holland J. J., Kiehn E. D. 1968; Specific cleavage of viral proteins as steps in the synthesis and maturation of enteroviruses. Proceedings of the National Academy of Sciences, U.S.A 60:1015–1022
    [Google Scholar]
  9. Jacobson M. F., Baltimore D. 1968; Polypeptide cleavages in the formation of poliovirus proteins. Proceedings of the National Academy of Sciences, U.S.A 61:77–84
    [Google Scholar]
  10. Jacobson M. F., Asso J., Baltimore D. 1970; Further evidence on the formation of poliovirus proteins. Journal of Molecular Biology 49:657–669
    [Google Scholar]
  11. Kitamura N., Semler B. L., Rothberg P. G., Larsen G. R., Adler C. J., Emini E. A., Hanecak R., Lee J. J., Van Der Werf S., Anderson C. W., Wimmer E. 1981; Primary structure, gene organization, and polypeptide expression of poliovirus RNA. Nature, London 291:547–553
    [Google Scholar]
  12. Kozak M. 1986; Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44:283–292
    [Google Scholar]
  13. Kyhse-Andersen J. 1984; Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of protein from polyacrylamide to nitrocellulose. Journal of Biochemical and Biophysical Methods 10:203
    [Google Scholar]
  14. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  15. Matsuura Y., Possee R. D., Overton H. A., Bishop D. H. L. 1987; Baculovirus expression vectors: the requirements for high level expression of proteins, including glycoproteins. Journal of General Virology 68:1233–1250
    [Google Scholar]
  16. Minor P. D. 1980; Comparative biochemical studies of type 3 poliovirus. Journal of Virology 34:73–84
    [Google Scholar]
  17. Minor P. D., Schild G. C., Bootman J., Evans D. M. A., Ferguson M., Reeve P., Spitz M., Stanway G., Cann A. J., Hauptmann R., Clarke L. D., Mountford R. C., Almond J. W. 1983; Location and primary structure of a major antigenic site for poliovirus neutralization. Nature, London 301:674–679
    [Google Scholar]
  18. Minor P. D., Evans D. M. A., Ferguson M., Schild G. C., Westrop G., Almond J. W. 1985; Principal and subsidiary antigenic sites of VP1 involved in the neutralization of poliovirus type 3. Journal of General Virology 66:1159–1165
    [Google Scholar]
  19. Minor P. D., Ferguson M., Evans D. M. A., Almond J. W., Icenogle J. P. 1986; Antigenic structure of poliovirus of serotypes 1, 2 and 3. Journal of General Virology 67:1283–1291
    [Google Scholar]
  20. Palmenberg A. C. 1982; In vitro synthesis and assembly of picornaviral capsid intermediate structures. Journal of Virology 44:900–906
    [Google Scholar]
  21. Racaniello V. R., Baltimore D. 1981; Molecular cloning of poliovirus cDNA and determination of the complete nucleotide sequence of the genome. Proceedings of the National Academy of Sciences, U.S.A 78:4887–4891
    [Google Scholar]
  22. Stanway G., Cann A. J., Hauptmann R., Hughes P., Clark L. D., Mountford R. C., Minor P. D., Schild G. C., Almond J. W. 1983; The nucleotide sequence of poliovirus type 3 Leon 12 a1b: comparison with poliovirus type 1. Nucleic Acids Research 11:5629–5643
    [Google Scholar]
  23. Stanway G., Mountford R. C., Cox S. D. J., Schild G. C., Minor P. D., Almond J. W. 1984; Molecular cloning of the genomes of poliovirus type 3 strains by the cDNA:RNA hybrid method. Archives of Virology 81:1539–1543
    [Google Scholar]
  24. Toyoda H., Kohara M., Kataoka Y., Suganuma T., Omata T., Imura N., Nomoto A. 1984; The complete nucleotide sequence of all three poliovirus serotype genomes: implication for the genetic relationship, gene function and antigenic determinants. Journal of Molecular Biology 174:561–585
    [Google Scholar]
  25. Toyoda H., Nicklin M. J. H., Murry M. G., Anderson C. W., Dunn J. J., Studier F. W., Wimmer E. 1986; A second virus-encoded proteinase involved in proteolytic processing of poliovirus polyprotein. Cell 45:761–770
    [Google Scholar]
  26. Van Der Werf S., Wychowski C, Bruneau P., Blondel B., Crainic R., Horodniceanu F., Girard M. 1983; Localization of a poliovirus type 1 neutralization epitope in viral capsid polypeptide VP1. Proceedings of the National Academy of Sciences, U.S.A 80:5080–5084
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-70-6-1453
Loading
/content/journal/jgv/10.1099/0022-1317-70-6-1453
Loading

Data & Media loading...

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