Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1992 May;66(5):2617–2630. doi: 10.1128/jvi.66.5.2617-2630.1992

Molecular attenuation of vaccinia virus: mutant generation and animal characterization.

M S Lee 1, J M Roos 1, L C McGuigan 1, K A Smith 1, N Cormier 1, L K Cohen 1, B E Roberts 1, L G Payne 1
PMCID: PMC241015  PMID: 1560521

Abstract

These studies demonstrated that the inbred BALB/c mouse strain can be optimized for the assessment of vaccinia virus virulence, growth, and spread from the site of inoculation and immune protection from a lethal vaccinia virus challenge. The studies established that manipulation of the vaccinia virus genome generated mutants exhibiting a wide range of attenuated phenotypes. The nine NYCBH vaccinia virus mutants had intracranial 50% lethal doses that ranged from 2 to greater than 7 log10 units. The decreased neurovirulence was due to decreased replication in brain tissue. Three mutants had a decreased ability to disseminate to the lungs, brains, livers, and spleens of mice after intranasal infection. One mutant had a decreased transmission from mice infected by tail scarification to naive cage mates. Although the mutants, with one exception, grew to wild-type titers in cell culture, they showed a growth potential on the scarified skin of mice that was dramatically different from that of the wild-type virus. Consequently, all of the mutants had significantly compromised immunogenicities at low virus immunization doses compared with that of the wild-type virus. Conversely, at high immunization doses most mutants could induce an immune response similar to that of the wild-type virus. Three Wyeth vaccine strain mutants were also studied. Whereas the thymidine kinase, ribonucleotide reductase, and hemagglutinin mutants had a reduced virulence (50% lethal dose), only the thymidine kinase mutant retained its immunogenicity.

Full text

PDF
2617

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Buller R. M., Chakrabarti S., Cooper J. A., Twardzik D. R., Moss B. Deletion of the vaccinia virus growth factor gene reduces virus virulence. J Virol. 1988 Mar;62(3):866–874. doi: 10.1128/jvi.62.3.866-874.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Buller R. M., Smith G. L., Cremer K., Notkins A. L., Moss B. Decreased virulence of recombinant vaccinia virus expression vectors is associated with a thymidine kinase-negative phenotype. 1985 Oct 31-Nov 6Nature. 317(6040):813–815. doi: 10.1038/317813a0. [DOI] [PubMed] [Google Scholar]
  3. Chakrabarti S., Brechling K., Moss B. Vaccinia virus expression vector: coexpression of beta-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol. 1985 Dec;5(12):3403–3409. doi: 10.1128/mcb.5.12.3403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Child S. J., Palumbo G. J., Buller R. M., Hruby D. E. Insertional inactivation of the large subunit of ribonucleotide reductase encoded by vaccinia virus is associated with reduced virulence in vivo. Virology. 1990 Feb;174(2):625–629. doi: 10.1016/0042-6822(90)90119-c. [DOI] [PubMed] [Google Scholar]
  5. Dallo S., Esteban M. Isolation and characterization of attenuated mutants of vaccinia virus. Virology. 1987 Aug;159(2):408–422. doi: 10.1016/0042-6822(87)90480-6. [DOI] [PubMed] [Google Scholar]
  6. Dallo S., Maa J. S., Rodriguez J. R., Rodriguez D., Esteban M. Humoral immune response elicited by highly attenuated variants of vaccinia virus and by an attenuated recombinant expressing HIV-1 envelope protein. Virology. 1989 Nov;173(1):323–329. doi: 10.1016/0042-6822(89)90250-x. [DOI] [PubMed] [Google Scholar]
  7. Gillard S., Spehner D., Drillien R. Mapping of a vaccinia host range sequence by insertion into the viral thymidine kinase gene. J Virol. 1985 Jan;53(1):316–318. doi: 10.1128/jvi.53.1.316-318.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ginsberg H. S., Lundholm-Beauchamp U., Horswood R. L., Pernis B., Wold W. S., Chanock R. M., Prince G. A. Role of early region 3 (E3) in pathogenesis of adenovirus disease. Proc Natl Acad Sci U S A. 1989 May;86(10):3823–3827. doi: 10.1073/pnas.86.10.3823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Goebel S. J., Johnson G. P., Perkus M. E., Davis S. W., Winslow J. P., Paoletti E. The complete DNA sequence of vaccinia virus. Virology. 1990 Nov;179(1):247-66, 517-63. doi: 10.1016/0042-6822(90)90294-2. [DOI] [PubMed] [Google Scholar]
  10. JOKLIK W. K. The purification fo four strains of poxvirus. Virology. 1962 Sep;18:9–18. doi: 10.1016/0042-6822(62)90172-1. [DOI] [PubMed] [Google Scholar]
  11. Javier R. T., Izumi K. M., Stevens J. G. Localization of a herpes simplex virus neurovirulence gene dissociated from high-titer virus replication in the brain. J Virol. 1988 Apr;62(4):1381–1387. doi: 10.1128/jvi.62.4.1381-1387.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kempe C. H., Fulginiti V., Minamitani M., Shinefield H. Smallpox vaccination of eczema patients with a strain of attenuated live vaccinia (CVI-78). Pediatrics. 1968 Dec;42(6):980–985. [PubMed] [Google Scholar]
  13. Kotwal G. J., Hügin A. W., Moss B. Mapping and insertional mutagenesis of a vaccinia virus gene encoding a 13,800-Da secreted protein. Virology. 1989 Aug;171(2):579–587. doi: 10.1016/0042-6822(89)90627-2. [DOI] [PubMed] [Google Scholar]
  14. Lane J. M., Ruben F. L., Neff J. M., Millar J. D. Complications of smallpox vaccination, 1968. N Engl J Med. 1969 Nov 27;281(22):1201–1208. doi: 10.1056/NEJM196911272812201. [DOI] [PubMed] [Google Scholar]
  15. Mackett M., Smith G. L., Moss B. Vaccinia virus: a selectable eukaryotic cloning and expression vector. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7415–7419. doi: 10.1073/pnas.79.23.7415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Morgan A. J., Mackett M., Finerty S., Arrand J. R., Scullion F. T., Epstein M. A. Recombinant vaccinia virus expressing Epstein-Barr virus glycoprotein gp340 protects cottontop tamarins against EB virus-induced malignant lymphomas. J Med Virol. 1988 Jun;25(2):189–195. doi: 10.1002/jmv.1890250209. [DOI] [PubMed] [Google Scholar]
  17. Morita M., Suzuki K., Yasuda A., Kojima A., Sugimoto M., Watanabe K., Kobayashi H., Kajima K., Hashizume S. Recombinant vaccinia virus LC16m0 or LC16m8 that expresses hepatitis B surface antigen while preserving the attenuation of the parental virus strain. Vaccine. 1987 Mar;5(1):65–70. doi: 10.1016/0264-410x(87)90012-0. [DOI] [PubMed] [Google Scholar]
  18. Panicali D., Grzelecki A., Huang C. Vaccinia virus vectors utilizing the beta-galactosidase assay for rapid selection of recombinant viruses and measurement of gene expression. Gene. 1986;47(2-3):193–199. doi: 10.1016/0378-1119(86)90063-6. [DOI] [PubMed] [Google Scholar]
  19. Panicali D., Paoletti E. Construction of poxviruses as cloning vectors: insertion of the thymidine kinase gene from herpes simplex virus into the DNA of infectious vaccinia virus. Proc Natl Acad Sci U S A. 1982 Aug;79(16):4927–4931. doi: 10.1073/pnas.79.16.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shida H., Hinuma Y., Hatanaka M., Morita M., Kidokoro M., Suzuki K., Maruyama T., Takahashi-Nishimaki F., Sugimoto M., Kitamura R. Effects and virulences of recombinant vaccinia viruses derived from attenuated strains that express the human T-cell leukemia virus type I envelope gene. J Virol. 1988 Dec;62(12):4474–4480. doi: 10.1128/jvi.62.12.4474-4480.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shida H., Tochikura T., Sato T., Konno T., Hirayoshi K., Seki M., Ito Y., Hatanaka M., Hinuma Y., Sugimoto M. Effect of the recombinant vaccinia viruses that express HTLV-I envelope gene on HTLV-I infection. EMBO J. 1987 Nov;6(11):3379–3384. doi: 10.1002/j.1460-2075.1987.tb02660.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Spyropoulos D. D., Roberts B. E., Panicali D. L., Cohen L. K. Delineation of the viral products of recombination in vaccinia virus-infected cells. J Virol. 1988 Mar;62(3):1046–1054. doi: 10.1128/jvi.62.3.1046-1054.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES