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Volume 76, Issue 11

Passage of Japanese encephalitis virus in HeLa cells results in attenuation of virulence in mice Free

Abstract

Of four wild-type strains (Nakayama-original, SA14, 826309 and Beijing-1) of Japanese encephalitis (JE) virus that were passaged six times in HeLa cells (HeLa p6), two (Nakayama-original and 826309) became attenuated for mice. In the case of strain Nakayama-original, the virulence for mice was markedly reduced and attenuation was retained on passage in primary chicken embryo fibroblast, LLC-MK2 and C6/36 cells. The binding of non-HeLa-passaged Nakayama virus to mouse brain membrane receptor preparations could be differentiated from binding by Nakayama HeLa p6 virus, suggesting that the envelope (E) protein is involved in the attenuated phenotype. Both of the attenuated viruses can be distinguished from the virulent non-HeLa-passaged parental viruses by examination with E protein reactive vaccine and wild-type-specific monoclonal antibodies (MAbs). The vaccine-specific MAb V23, which is only reactive with the SA14 series of live vaccine viruses, recognized the HeLa cell-attenuated Nakayama-original and 826309 viruses, whereas two wild-type-specific MAbs (MAbs K13 and K39) lost reactivity. Comparison of the nucleotide sequences of the structural protein genes of the 826309 and Nakayama-original virulent parent and attenuated HeLa p6 viruses revealed that the viruses differed by 37 and 46 nucleotides coding for eight and nine amino acid mutations, respectively. However, other than one amino acid in the E protein, the membrane and E protein amino acid sequences of the two attenuated HeLa p6 viruses were identical.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1995
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1995-11-01
2024-04-24
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References

  1. Aihara S., Chunming R., Yu Y. X., Lee T., Watanbe K., Komiya T., Sumiyoshi H., Hashimoto H., Nomoto A. 1991; Identification of mutations that occurred on the genome of Japanese encephalitis virus during the attenuation process. Virus Genes 5:95–109
     [Google Scholar]
  2. Barrett A. D. T., Monath T. P., Cropp C. B., Adkins J. A., Ledger T. N., Gould E. A., Schlesinger J. J., Kinney R. M., Trent D. W. 1990; Attenuation of wild-type yellow fever virus by passage in HeLa cells. Journal of General Virology 71:2301–2306
     [Google Scholar]
  3. Cecilia D., Gould E. A. 1991; Nucleotide changes responsible for loss of neuroinvasiveness in Japanese encephalitis virus neutralization-resistant mutants. Virology 181:70–77
     [Google Scholar]
  4. Dunster L. M., Gibson C. A., Stephenson J. R., Monor P. D., Barrett A. D. T. 1990; Attenuation of virulence of flaviviruses following passage in HeLa cells. Journal of General Virology 71:601–607
     [Google Scholar]
  5. Hardy F. M. 1963; The growth of Asibi strain yellow fever virus in tissue culture. II: modification of virus and cells. Journal of Infectious Diseases 113:9–14
     [Google Scholar]
  6. Hasegawa H., Yoshida M., Shiosaka T., Fujita S., Kobayashi Y. 1992; Mutations in the envelope protein of Japanese encephalitis virus affect entry into cultured cells and virulence in mice. Virology 191:158–165
     [Google Scholar]
  7. Hashimoto H., Nomoto A., Watanabe K., Mori T., Takezawa T., Aizawa C., Takegami T., Hirarnatsu K. 1988; Molecular cloning and complete nucleotide sequence of the genome of Japanese encephalitis virus Beijing-1 strain. Virus Genes 1:305–317
     [Google Scholar]
  8. Higgins D. G., Sharp P. M. 1988; CLUSTAL: a package for performing multiple sequence alignments on a microcomputer. Gene 73:237–244
     [Google Scholar]
  9. Huang C. H. 1982; Studies of Japanese encephalitis in China. Advances in Virus Research 27:71–101
     [Google Scholar]
  10. McAda P. C., Mason P. W., Schmaljohn C. S., Dalrymple J. M., Mason T. L., Fournier M. J. 1987; Partial nucleotide sequence of the Japanese encephalitis virus genome. Virology 158:348–360
     [Google Scholar]
  11. Middlemiss D. N., Fozard J. R. 1983; 80HDPAT discrimination between subtypes of the 5-HT1 recognition site. European Journal of Pharmacology 90:151–153
     [Google Scholar]
  12. Miller B. R., Adkins D. 1988; Biological characterization of plaque-size variants of yellow fever virus in mosquitos and mice. Acta Virologica 32:227–234
     [Google Scholar]
  13. Ni H., Burns N. J., Chang G.-J., Zhang M.-J., Wills M. R., Trent D. W., Sanders P. G., Barrett A. D. T. 1994; Comparison of nucleotide and deduced amino acid sequence of the structural protein genes of the wild-type Japanese encephalitis virus strain SAM and its attenuated vaccine derivatives. Journal of General Virology 75:1505–1510
     [Google Scholar]
  14. Ni H., Chang G.-J., Xie H., Trent D. W., Barrett A. D. T. 1995; Molecular basis of attenuation of neuro virulence of wild-type Japanese encephalitis virus strain SAM. Journal of General Virology 76:409–413
     [Google Scholar]
  15. Nitayaphan S., Chang G. G., Trent D. W. 1990; Nucleotide sequence of the virulent SA-14 strain of Japanese encephalitis virus and its attenuated vaccine derivative, SA-14-14-2. Virology 111:541–552
     [Google Scholar]
  16. Queen C. G., Korn L. J. 1984; A comprehensive sequence analysis program for the IBM personal computer. Nucleic Acids Research 12:581–599
     [Google Scholar]
  17. Rosen L. 1986; The natural history of Japanese encephalitis virus. Annual Reviews of Microbiology 40:395–409
     [Google Scholar]
  18. Sil B. K., Wills M. R., Cao J. X., Sharda R., Islam M. A., Stagg D. J., Jennings A. D., Gibson C. A., Barrett A. D. T. 1992a; Immunogenicity of experimental live attenuated Japanese encephalitis vaccine viruses and comparison with wild-type strains using monoclonal and polyclonal antibodies. Vaccine 10:329–333
     [Google Scholar]
  19. Sil B. K., Dunster L. M., Ledger T. N., Wills M. R., Minor P. D., Barrett A. D. T. 1992b; Identification of envelope protein epitopes that are important in the attenuation process of wild-type yellow fever virus. Journal of Virology 66:4265–4270
     [Google Scholar]
  20. Sumiyoshi H., Mori C., Fuke I., Morita K., Kuhara S., Kondou J., Kikuchi Y., Nagamatsu H., Igarashi A. 1987; Complete nucleotide sequence of the Japanese encephalitis virus genome RNA. Virology 161:497–510
     [Google Scholar]
  21. Wills M. R., Sil B. K., Cao J. X., Yu Y. X., Barrett A. D. T. 1992; Antigenic characterization of the live attenuated Japanese encephalitis vaccine virus SA14-14-2: a comparison with isolates of the virus covering a wide geographic area. Vaccine 10:861–872
     [Google Scholar]
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Passage of Japanese encephalitis virus in HeLa cells results in attenuation of virulence in mice
J. Gen. Virol. 76, 2757 (1995); https://doi.org/10.1099/0022-1317-76-11-2757
/content/journal/jgv/10.1099/0022-1317-76-11-2757
/content/journal/jgv/10.1099/0022-1317-76-11-2757
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