Construction and Characterization of Deletion Mutants of Pseudorabies Virus: a New Generation of ‘Live’ Vaccines Free

Abstract

SUMMARY

Various deletions were introduced into a cloned subgenomic fragment (HI-1), located in the unique short (U) region of the DNA from the virulent Northern Ireland Aujeszky-3 (NIA-3) strain of pseudorabies virus (PRV). In the cloned dIII-B fragment, the I -II fragment was replaced by different I-II fragments of the deleted HI-7clones. Transfection of the deleted dIII-B fragments together with the dIII-A fragment of either the NIA-3 or the non-virulent NIA-4 strain yielded replication-competent deletion mutants. The region in U in which sequences were deleted specified several mRNAs. Some of the mRNAs present in cells infected with NIA-3 were absent from cells infected with the deletion mutants, whereas other differently sized mRNAs were generated. The mutants were examined with respect to their biological properties in cell culture, mice and pigs. The results showed that (i) the type of cytopathic effect induced in cell culture seemed to be determined by the Uregion, (ii) using the mean time to death in mice as a parameter, markers for virulence were present in the U and U regions and (ii) the introduction of deletions in Ustrongly reduced the virulence of PRV for pigs. Despite the impaired capacity of the deletion mutants to induce high titres of neutralizing antibodies in the serum, inoculation with mutants derived from NIA-3 prevented clinical disease in pigs upon challenge with the virulent parent strain. These deletion mutants provide a good basis for the production of bioengineered live PRV vaccines.

Keyword(s): deletion mutants , PRV and vaccine
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1987-02-01
2024-03-29
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References

  1. Baskerville A., Mcferran J. B., C. Dow. 1973; Aujeszky’s disease in pigs. Veterinary Bulletin 43:465–480
    [Google Scholar]
  2. Ben-PORAT T., Rixon F. J., Blankenship M. L. 1979; Analysis of the structure of the genome of pseudorabies virus. Virology 95:285–294
    [Google Scholar]
  3. Ben-Porat T., Demarche J., Pendrys J., Veach R. A., Kaplan A. S. 1986; Proteins specified by the short unique region of the genome of pseudorabies virus play a role in the release of virions from certain cells. Journal of Virology 57:191–196
    [Google Scholar]
  4. Berns A., Vanderouweland A., Quint W., Vanoirschot J., Gielkens A. 1985; Presence of markers for virulence in the unique short region or repeat region or both of pseudorabies hybrid viruses. Journal of Virology 53:89–93
    [Google Scholar]
  5. Bitsch v. 1980; Correlation between the pathogenicity of field strains of Aujeszky’s disease virus and their ability to cause cell fusion - syncytia formation - in cell cultureS. Acta veterinaria scandinavica 21:708–710
    [Google Scholar]
  6. Bitsch v., Eskildsen M. 1976; A comparative examination of swine sera for antibody to Aujeszky virus with the conventional test and a modified virus-serum neutralization test and a modified direct complement fixation test. Acta veterinaria scandinavica 17:142–152
    [Google Scholar]
  7. Deleeuw p. W., Vanoirschot J. T. 1985; Vaccines against Aujeszky’s disease: evaluation of their efficacy under standardized laboratory conditionS. Veterinary Quarterly 7:191–197
    [Google Scholar]
  8. Deleeuw p. W., Wijsmuller J. M., Zantinga J. W., Tielen M. J. M. 1982; Intranasal vaccination of pigs against Aujeszky’s disease. 1. Comparison of intranasal or parenteral vaccination with an attenuated vaccine in 12 week old pigs from immunized dams. Veterinary Quarterly 4:49–56
    [Google Scholar]
  9. Gielkens A., Berns A. 1982; Differentiation of Aujeszky’s disease virus strains by restriction endonuclease analysis of the viral DNAs. Current Topics in Veterinary Medicine and Animal Science 17:3–13
    [Google Scholar]
  10. Gielkens A. L. J., Briaire l. 1984; Genome differences among plaque isolates of Aujeszky’s disease virus vaccines. Abstract 8 th International Pig Veterinary Society Congress (Ghent) p 19
    [Google Scholar]
  11. Gielkens A. L. J., Vanoirschot J. T., Berns A. J. M. 1985; Genome differences among field isolates and vaccine strains of pseudorabies virus. Journal of General Virology 66:69–82
    [Google Scholar]
  12. Graham F. L., Vandereb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–467
    [Google Scholar]
  13. Honess R. W., Watson D. H. 1977; Unity and diversity in the herpesviruses. Journal of General Virology 37:15–37
    [Google Scholar]
  14. Kaplan A. S. 1969; Herpes simplex and pseudorabies viruses. Virology Monographs 5 Gard S., Hallauer C., Meyer K. F. Edited by Wien & New York: Springer-Verlag;
    [Google Scholar]
  15. Ladin B. F., Ihara S., Hampl H., Ben-PORAT T. 1982; Pathway of assembly of herpesvirus capsids: an analysis using DNA+ temperature-sensitive mutants of pseudorabies virus. Virology 116:544–561
    [Google Scholar]
  16. Lomniczi B., Blankenship M. L., Ben-PORAT T. 1984; Deletions in the genomes of pseudorabies virus vaccine strains and existence of four isomers of the genome. Journal of Virology 49:970–979
    [Google Scholar]
  17. Mettenleiter T. C., Lukacs N., Rziha H. J. 1985a; Mapping of the structural gene of pseudorabies virus glycoprotein A and identification of two non-glycosylated precursor polypeptides. Journal of Virology 53:52–57
    [Google Scholar]
  18. Mettenleiter T. C., Lukacs N., Rziha H. J. 1985b; Pseudorabies virus avirulent strains fail to express a major glycoprotein. Journal of Virology 56:307–311
    [Google Scholar]
  19. Platt K. B., Mare C. J., Hinz p. N. 1980; Differentiation of vaccine strains and of field isolates of pseudorabies (Aujeszky’s disease) virus: trypsin sensitivity and mouse virulence markers. Archives of Virology 63:107–114
    [Google Scholar]
  20. Quint W., Quax W., Vanderputten H., Berns A. 1981; Characterization of AKR murine leukemia virus sequences in AKR mouse substrains and structure of integrated recombinant genomes in tumor tissues. Journal of Virology 39:1–10
    [Google Scholar]
  21. Rixon F. J., Ben-PORAT T. 1979; Structural evolution of the DNA of pseudorabies defective viral particles. Virology 97:151–163
    [Google Scholar]
  22. Roizman B., Jacob R. J., Knipe D. M., Morse L. S., Ruyechan W. T. 1978; On the structure, functional equivalence, and replication of the four arrangements of herpes simplex virus DNA. Cold Spring Harbor Symposia on Quantitative Biology 43:809–826
    [Google Scholar]
  23. Rubenstein A. S., Kaplan A. S. 1975; Electron microscopic studies of the DNA of defective and standard pseudorabies virions. Virology 66:385–392
    [Google Scholar]
  24. Thomas P. S. 1980; Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proceedings of the National Academy of Sciences, U.S.A. 77:5201–5205
    [Google Scholar]
  25. Vanderputten H., Quint W., Vanraay J., Robanusmaandag E., Verma I. M., Berns A. 1981; M-MuLV induced leukemogenesis: integration and structure of recombinant proviruses in tumors. Cell 24:729–739
    [Google Scholar]
  26. Vanoirschot J. T., Gielkens A. L. J. 1984a; Some characteristics of four attenuated vaccine virus strains and a virulent strain of Aujeszky’s disease virus. Veterinary Quarterly 6:225–229
    [Google Scholar]
  27. Vanoirschot J. T. 1984b; Intranasal vaccination of pigs against pseudorabieS. 2. Absence of vaccinal virus latency and failure to prevent latency of virulent virus. American Journal of Veterinary Research 45:2099–2103
    [Google Scholar]
  28. Vanoirschot J. T., Quint W., Gielkens A. L. J., Deleeuw P. W., Berns A. J. M. 1985; On the immunity induced in pigs by conventional vaccines and deletion mutants of Aujeszky’s disease virus. Proceedings, EEC Seminar on Immunity to Herpesvirus Infections of Domestic Animals (Brussels, December 4-5 1984) pp. 281–290
    [Google Scholar]
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