Журнал микробиологии, эпидемиологии и иммунобиологии. 2019; : 68-76
Изучение биологических свойств аттенуированных вариантов штамма А/WSN/33, полученных с помощью сайт-специфического мутагенеза РВ2-гена
https://doi.org/10.36233/0372-9311-2019-2-68-76Аннотация
Цель. Изучение биологических свойств аттенуированных вариантов штамма А/WSN/33(Н1N1) вируса гриппа А, полученных с помощью сайт-специфического мутагенеза РВ2-гена. Материалы и методы. С помощью методов обратной генетики получены сайт-специфические мутанты штамма А/WSN/33, имеющие в РВ2-гене ts-мутации из генома холодоадаптированных (ХА) штаммов-доноров аттенуации: А/Энн Арбор/6/60(Н2N2), А/Ленинград/134/17/57(Н2N2), А/Краснодар/101/35/59 (Н2N2). У полученных сайт-специфических мутантов исследован ts-фенотип, att-фенотип, иммуногенность и защитная эффективность при гомологичном и гетерологичном контрольном заражении. Результаты. Показано, что включение в РВ2-ген вирулентного штамма А/WSN/33 как единичных ts-мутаций, так и комбинации ts-мутаций из генома известных ХА штаммов-доноров аттенуации ведет к изменению ts- и att-фенотипа полученных сайт-специфических мутантов. Наблюдалось падение способности к размножению при повышенной температуре и снижение вирулентности для мышей при интраназальном заражении. Полученные мутанты имели высокую защитную эффективность при гомологическом и гетерологическом контрольном заражении. Заключение. Полученные данные позволяют сделать вывод, что некоторые сайт-специфические мутанты не уступают по защитной эффективности как при гомологичном, так и гетерологичном контрольном заражении ХА реассортантным вакцинным вариантам. Результаты работы дают основания рассматривать некоторые из этих мутантов как возможные кандидаты в живые гриппозные вакцины.
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16. Solorzano A.,Li Yo., Perez D.R. Alternative live —attenuated influenza vaccines based on modification in the polymerase genes protect against epidemic and pandemic flu. J. Virol. 2010, 84(9):4587-4596.
17. Song H., Nieto G., Perez D. A new generation of modified live-attenuated avian influenza viruses using a two-strategy combination as potential vaccine candidates. J. Virol. 2007, 81(17):9238-9248.
18. Subbarao E.K., Park E., Lawson C. et al. Secvential addition of temperature-sensitive missense mutations into the PB2 gene of influenza A transfectant viruses can effect an increase in temperature sensitivity and attenuation and permits the rational design of a genetically engineered live influenza A virus vaccine. J. Virol. 1995, 69(10):59-69.
19. Yamanaka K., Ogasawara N., Ueda M. et al. Characterization of a temperature —sensitive mutant in the RNA polymerase PB2 subunit gene of influenza A/WSN/33 virus. Arch. Virol. 1990, 114:65-73.
20. Zhou B., Li Yo., Speer S. et al. Engineering temperature sensitive live attenuated influenza vaccines from emerging viruses. Vaccine. 2012, 30(24):3691-3702.
Journal of microbiology, epidemiology and immunobiology. 2019; : 68-76
Study of the biological properties of attenuated variants of the virulent A/WSN/33 strain of influenza virus, obtained by the site-specific mutagenesis of PB2-gene
https://doi.org/10.36233/0372-9311-2019-2-68-76Abstract
Aim. Study of biological properties of attenuated variants of the virulent A/WSN/33 strain of influenza virus, obtained by the site-specific mutagenesis of PB2-gene. Materials and methods. Site-specific mutants of A/WSN/33 of influenza virus, having in PB2-gene ts-mutations from genome of cold-adapted (CA) master-strains: A/Ann Arbor/6/60 (H2N2); A/Leningrad/134/17/57 (H2N2); A/Krasnodar/101/35/59 (H2N2) were obtained with help of reverse genetics methods. The ts-phenotype, att-phenotype, immunogenicity and protective efficacy in homologous and heterologous control infections were studied in the obtained site-specific mutants. Results. It was shown that the inclusion in the PB2-gene of the virulent A/WSN/33 strain as single mutations and a combination of mutations from the genomes of CA donor-strains leads to a change in the ts-phenotype and att-phenotype of the mutants obtained. These mutants had high protective efficacy in homologous and heterologous control infection. Conclusion. The results obtained allow us to consider the site-specific mutants of influenza virus as possible candidates for live influenza vaccines.
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7. Jin H., Zhou H., Lu B.et al. Imparting temperature sensitivity and attenuation in ferrets to A/Puerto Rico/8/34 influenza virus by transferring the genetic signature for temperature sensitivity from cold-adapted A/Ann Arbor/6/60. J. Virol. 2004, 78 (2):995-998.
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9. Lawson C.M., Subbarao E.K., Murphy B.R. Nucleotide sequences changes in the polymerase basic protein 2 gene of temperature-sensitive mutants of influenza A virus. Virology.1992, 191:506-510.
10. Maassab H. Adaptation and growth characteristics of influenza virus at 25 degrees C. Nature. 1967, 213:612-614.
11. McCauley J.W., Penn C.R. The critical cut-off temperature of avian influenza viruses. Virus Res. 1990, 17:191-198.
12. Parkin N., Chiu P., Coelingh K. Genetically engineering live attenuated influenza A virus vaccine candidates. J. Virol. 1997, 71(4):2772-2778.
13. Poleshaev F.I. The conditions of influenza virus ts-recombinants development. Acta virologica. 1978, 22:263-269.
14. Smirnov Y.A. et al. Characterization of adaptation of an avian influenza A (H5N2) virus to mammalian host. Acta virologica. 2000, 44(1):1-8.
15. Snyder M. H., Betts R.F.,De Borde D. et al. Four viral genes independently contribute to attenuation of live influenza A/Ann Arbor/6/60 (H2N2) cold-adapted reassortant virus vaccines. J. Virol. 1988, 62:488-495.
16. Solorzano A.,Li Yo., Perez D.R. Alternative live —attenuated influenza vaccines based on modification in the polymerase genes protect against epidemic and pandemic flu. J. Virol. 2010, 84(9):4587-4596.
17. Song H., Nieto G., Perez D. A new generation of modified live-attenuated avian influenza viruses using a two-strategy combination as potential vaccine candidates. J. Virol. 2007, 81(17):9238-9248.
18. Subbarao E.K., Park E., Lawson C. et al. Secvential addition of temperature-sensitive missense mutations into the PB2 gene of influenza A transfectant viruses can effect an increase in temperature sensitivity and attenuation and permits the rational design of a genetically engineered live influenza A virus vaccine. J. Virol. 1995, 69(10):59-69.
19. Yamanaka K., Ogasawara N., Ueda M. et al. Characterization of a temperature —sensitive mutant in the RNA polymerase PB2 subunit gene of influenza A/WSN/33 virus. Arch. Virol. 1990, 114:65-73.
20. Zhou B., Li Yo., Speer S. et al. Engineering temperature sensitive live attenuated influenza vaccines from emerging viruses. Vaccine. 2012, 30(24):3691-3702.
