vestibИзвестия Национальной академии наук Беларуси. Серия биологических наукProceedings of the National Academy of Sciences of Belarus, Biological Series1029-89402524-230XThe Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"10.29235/1029-8940-2018-63-4-419-425vestib-393Research ArticleСтатьиГенетическая конструкция для создания ДНК-вакцины против репродуктивно-респираторного синдрома свинейDesign of genetic construction for creation DNA vaccine against porcine reproductive and respiratory syndromeКравченкоЛ. М.KravchenkoL. M.

Кравченко Лидия Михайловна – младший научный сотрудник.

пр. Независимости,  4,  220030, Минск.

Lidia M. Kravchenko – Junior researcher.

4, Nezavisimosti Ave., 220030, Minsk.

lidia_kravchenko@tut.byКудинК. В.KudzinK. V.

Кудин Кирилл Валерьевич – научный сотрудник.

пр. Независимости, 4, 220030, Минск.

Kiryl V. Kudzin – Researcher.

4, Nezavisimosti Ave., 220030, Minsk.

kiryl.kudin@gmail.comПрокулевичВ. А.PrakulevichU. A.

Прокулевич Владимир Антонович – доктор биологических наук, профессор, заведующий кафедрой.

пр. Независимости, 4, 220030, Минск.

Uladzimir A. Prakulevich – D. Sc. (Biol.), Professor, Head of the Department.

4, Nezavisimosti  Ave.,  220030,  Minsk.

prokulevich@mail.ruБелорусский государственный университетBelarusian State University201830102018634419425Copyright © Кравченко Л.М., Кудин К.В., Прокулевич В.А., 20182018Кравченко Л.М., Кудин К.В., Прокулевич В.А.Kravchenko L.M., Kudzin K.V., Prakulevich U.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestibio.belnauka.by/jour/article/view/393

Серьезный экономический ущерб свиноводству по всему миру наносит репродуктивно-респираторный синдром свиней (РРСС). Это вирусное инфекционное заболевание, против которого живые ослабленные и инактивированные вакцины не всегда успешны. В связи с этим надежды на повышение эффективности профилактики данного заболевания связывают с разработкой нового типа препаратов – ДНК-вакцин, которые способны индуцировать развитие как клеточного, так и гуморального иммунного ответа. Такие вакцины состоят из плазмидного или вирусного вектора, в который встроены гены потенциально иммуногенных белков возбудителя инфекционного заболевания. Экспрессия этих генов осуществляется в клетках вакцинированного животного, что приводит к синтезу белков-антигенов, которые индуцируют запуск иммунного ответа.

Представленная в данной работе генетическая конструкция, разработанная на основе коммерческого плазмидного вектора pVAX1, может быть использована для создания ДНК-вакцины против РРСС. Модифицированный вектор содержит   открытые рамки считывания двух структурных белков вируса РРСС, участок гена инвариантной цепи, кодирующий сигнал лизосомной локализации, а также регуляторные элементы, необходимые для экспрессии клонированных генов в клетках млекопитающих.

The porcine reproductive and respiratory syndrome (PRRS) caused the serious economic damage to swine breeding around the world. It is a viral infective disease against which live attenuated and inactivated vaccines are not always successful. Development of new types of drugs such as DNA vaccines is necessary for improving the protection against the virus. DNA vaccines induce the development of both a cellular and humoral immune response. Such vaccines consist of a plasmid or viral vector with genes of potentially immunogenic proteins. The expression of these genes realized in cells of the vaccinated animal. It leads to the synthesis of antigen proteins triggering the immune response. The purpose of this work is to create a genetic construction that can be used as DNA vaccine against PRRS virus. The construction consists of the commercial vector pVAX1 and open reading frame of two structural proteins of PRRS virus, a lysosomal localization signal sequence of the invariant chain gene and regulatory elements necessary for the expression of cloned genes in mammalian cells.

репродуктивно-респираторный синдром свинейДНК-вакцинаинвариантная цепьMHC IIгенетическая конструкцияporcine reproductive and respiratory syndromeDNA vaccineinvariant chainMHC IIgenetic constructionReferencesLiu, M. A. DNA vaccines: a review / M. A Liu // J. Int. Med. – 2003. – Vol. 253, N 4. – P. 402–410.Liu M. A. DNA vaccines: a review. Journal of Internal Medicine, 2003, vol. 253, no. 4, pp. 402–410.Shedlock, D. J. DNA vaccination: antigen presentation and the induction of immunity / D. J. Shedlock, D. B. Weiner // J. Leuk. Biol. – 2000. – Vol. 68, N 6. – Р. 793–806.Shedlock D. J., Weiner D. B. DNA vaccination: antigen presentation and the induction of immunity. Journal of Leukocyte Biology, 2000, vol. 68, no. 6, pp. 793–806.Molecular and cellular mechanisms of DNA vaccines / C. Coban [et al.] // Human Vaccines. – 2008. – Vol. 4, N 6. – Р. 453–457. https://doi.org/10.4161/hv.4.6.6200Coban C., Koyama S., Takeshita F., Akira S., Ishii K. J. Molecular and cellular mechanisms of DNA vaccines. Human Vaccines, 2008, vol. 4, no. 6, pp. 453–457. https://doi.org/10.4161/hv.4.6.6200Impact of genetic diversity of European-type porcine reproductive and respiratory syndrome virus strains on vaccine efficacy / G. Labarque [et al.] // Vaccine. – 2004. – Vol. 22, N 31–32. – P. 4183–4190. https://doi.org/10.1016/j.vaccine.2004.05.008Labarque G., van Reeth K., Nauwynck H., Drexler C., van Gucht S., Pensaert M. Impact of genetic diversity of European-type porcine reproductive and respiratory syndrome virus strains on vaccine efficacy. Vaccine, 2004, vol. 22, no. 31–32, pp. 4183–4190. https://doi.org/10.1016/j.vaccine.2004.05.008Failure of an inactivated vaccine against porcine reproductive and respiratory syndrome to protect gilts against a heterologous challenge with PRRSV / M. Scortti [et al.] // Vet. Rec. – 2007. – Vol. 161, N 24. – P. 809–813.Scortti M., Prieto C., Alvarez E., Simarro I., Castro J. M. Failure of an inactivated vaccine against porcine reproductive and respiratory syndrome to protect gilts against a heterologous challenge with PRRSV. Veterinary Record, 2007, vol. 161, no. 24, pp. 809–813.Evaluation of different DNA vaccines against porcine reproductive and respiratory syndrome (PRRS) in pigs / S. Petrini [et al.] // Vaccines. – 2013. – Vol. 1, N 4. – P. 463–480. https://doi.org/10.3390/vaccines1040463Petrini S., Ramadori G., Villa R., Borghetti P., de Angelis E., Cantoni A. M., Corradi A., Amici A., Ferrari M. Evaluation of different DNA vaccines against porcine reproductive and respiratory syndrome (PRRS) in pigs. Vaccines, 2013, vol. 1, no. 4, pp. 463–480. https://doi.org/10.3390/vaccines1040463Production and evaluation of virus-like particles displaying immunogenic epitopes of porcine reproductive and respiratory syndrome virus (PRRSV) / A. Murthy [et al.] // Int. J. Mol. Sci. – 2015. – Vol. 16, N 12. – P. 8382–8396. https://doi.org/10.3390/ijms16048382Murthy A., Ni Y., Meng X., Zhang Ch. Production and evaluation of virus-like particles displaying immunogenic epitopes of porcine reproductive and respiratory syndrome virus (PRRSV). International Journal of Molecular Sciences, 2015, vol. 16, no. 12, pp. 8382–8396. https://doi.org/10.3390/ijms16048382Lu, Z. H. Beyond the whole genome consensus: unravelling of PRRSV phylogenomics using next generation sequencing technologies / Z. H. Lu, A. L. Archibald, T. Ait-Ali // Virus Res. – 2014. – Vol. 194. – P. 167–174. https://doi.org/10.1016/j.virusres.2014.10.004Lu Z. H., Archibald A. L., Ait-Ali T. Beyond the whole genome consensus: unravelling of PRRSV phylogenomics using next generation sequencing technologies. Virus Research, 2014, vol. 194, pp. 167–174. https://doi.org/10.1016/j.virusres.2014.10.004Rossow, K. D. Porcine reproductive and respiratory syndrome / K. D. Rossow // Veterinary Pathology. – 1998. – Vol. 35, N 1. – P. 1–20.Rossow K. D. Porcine reproductive and respiratory syndrome. Veterinary Pathology, 1998, vol. 35, no. 1, pp. 1–20.Wang, R. Induction of antigen-specific cytotoxic t lymphocytes in humans by a malaria DNA vaccine / R. Wang // Science. – 1998. – Vol. 282, N 5388. – P. 476–480. https://doi.org/10.1126/science.282.5388.476Wang R. Induction of antigen-specific cytotoxic t lymphocytes in humans by a malaria DNA vaccine. Science, 1998, vol. 282, no. 5388, pp. 476–480. https://doi.org/10.1126/science.282.5388.476MHC class II transport from lysosomal compartments to the cell surface is determined by stable peptide binding, but not by the cytosolic domains of the β- and α-chains / C. Théry [et al.] // J. Immun. – 1998. – Vol. 161, N 5. – P. 2106–2113.Théry C., Brachet V., Regnault A., Rescigno M., Ricciardi-Castagnoli P., Bonnerot C., Amigorena S. MHC class II transport from lysosomal compartments to the cell surface is determined by stable peptide binding, but not by the cytosolic domains of the β- and α-chains. Journal of Immunology, 1998, vol. 161, no. 5, pp. 2106–2113.Встраивание сигнала направления в лизосомы инвариантной цепи изменяет деградацию обратной транскриптазы ВИЧ-1, повышая ее иммуногенность / Е. С. Стародубова [и др.] // Acta Naturae. – 2014. – Т. 6, № 1. – С. 66–73.Starodubova E. S., Isaguliants М. G., Kuzmenko Y. V., Latanova A. A., Krotova О. А., Karpov V. L. Fusion to the lysosome targeting signal of the invariant chain alters the processing and enhances the immunogenicity of HIV-1 reverse transcriptase. Acta Naturae, 2014, vol. 6, no. 1, pp. 61–68.Current protocols in molecular biology / ed. : F. M. Ausubel [et al.]. – 2nd ed. – New York : John Wiley & Sons, 1993. – 4410 p.Ausubel F. M., Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., Struhl K. (eds.). Current protocols in molecular biology. 2nd ed. New York, John Wiley & Sons, 1993. 4410 p.Immune responses in mice vaccinated with virus-like particle composed of the GP5 and M proteins of porcine reproductive and respiratory syndrome virus / H.-M. Nam [et al.] // Arch. Virol. – 2013. – Vol. 158, N 6. – P. 1275–1285. https://doi.org/10.1007/s00705-013-1612-zNam H.-M., Chae K.-S., Song Y.-J., Lee N.-H., Lee J.-B., Park S.-Y., Song Ch.-S., Seo K.-H., Kang S.-M., Kim M.-Ch., Choi I.-S. Immune responses in mice vaccinated with virus-like particle composed of the GP5 and M proteins of porcine reproductive and respiratory syndrome virus. Archives of Virology, 2013, vol. 158, no. 6, pp. 1275–1285. https://doi.org/10.1007/s00705-013-1612-zWootton, S. K. Antigenic structure of the nucleocapsid protein of porcine reproductive and respiratory syndrome virus / S. K. Wootton, E. A. Nelson, D. Yoo // Clin. Diagn. Lab. Immunol. – 1998. – Vol. 5, N 6. – P. 773–779.Wootton S. K., Nelson E. A., Yoo D. Antigenic structure of the nucleocapsid protein of porcine reproductive and respiratory syndrome virus. Clinical and Diagnostic Laboratory Immunology, 1998, vol. 5, no. 6, pp. 773–779.Immune responses in pigs induced by recombinant DNA vaccine co-expressing swine IL-18 and membrane protein of porcine reproductive and respiratory syndrome virus / X. Zhang [et al.] // Int. J. Mol. Sci. – 2012. – Vol. 13, N 5. – P. 5715–5728. https://doi.org/10.3390/ijms13055715Zhang X., Wang X., Mu L., Ding Z. Immune responses in pigs induced by recombinant DNA vaccine co-expressing swine IL-18 and membrane protein of porcine reproductive and respiratory syndrome virus. International Journal of Molecular Sciences, 2012, vol. 13, no. 5, pp. 5715–5728. https://doi.org/10.3390/ijms13055715Improved immunogenicity of DNA constructs co-expressing the GP5 and M proteins of porcine reproductive and respiratory syndrome virus by glycine-proline-glycine-proline (GPGP) linker in mice / M.-Y. Chia [et al.] // Taiwan Vet. J. – 2011. – Vol. 37, N 1. – Р. 12–23.Chia M.-Y., Hsiao S.-H., Chan H.-T., Do Y.-Y., Huang P.-L., Chang H.-W., Tsai Y.-C., Lin C.-M., Cheng C.-H., Pang V. F., Jeng C.-R. Improved immunogenicity of DNA constructs co-expressing the GP5 and M proteins of porcine reproductive and respiratory syndrome virus by glycine-proline-glycine-proline (GPGP) linker in mice. Taiwan Veterinary Journal, 2011, vol. 37, no. 1, pp. 12–23.Co-expressing GP5 and M proteins under different promoters in recombinant modified vaccinia virus Ankara (rMVA)-based vaccine vector enhanced the humoral and cellular immune responses of porcine reproductive and respiratory syndrome virus (PRRSV) / Q. Zheng [et al.] // Virus Genes. – 2007. – Vol. 35, N 3. – P. 585–595. https://doi.org/10.1007/s11262-007-0161-5Zheng Q., Chen D., Li P., Bi Z., Cao R., Zhou B., Chen P. Co-expressing GP5 and M proteins under different promoters in recombinant modified vaccinia virus Ankara (rMVA)-based vaccine vector enhanced the humoral and cellular immune responses of porcine reproductive and respiratory syndrome virus (PRRSV). Virus Genes, 2007, vol. 35, no. 3, pp. 585–595. https://doi.org/10.1007/s11262-007-0161-5

The authors declare that there are no conflicts of interest present.