Problems of VirologyProblems of VirologyВопросы вирусологии0507-40882411-2097Central Research Institute for Epidemiology1315910.36233/0507-4088-183pvluewORIGINAL RESEARCHESОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯResearch ArticleThe adjuvant effect of polymuramil, a NOD1 and NOD2 agonist, differs when immunizing mice of different inbred lines with nonstructural hepatitis C virus (Flaviviridae: Hepacivirus)proteins and is synergistically enhanced in combination with pyrogenalum, a TLR4 agonistАдъювантное действие агониста NOD1 и NOD2 полимурамила различается при иммунизации мышей разных инбредных линий неструктурными белками вируса гепатита С (Flaviviridae: Hepacivirus) и синергетически усиливается в комбинации с агонистом TLR4 пирогеналомhttps://orcid.org/0000-0002-2801-6843LesnovaEkaterina I.ЛесноваЕкатерина Ивановна
Russian Federation

Researcher

научный сотр.

wolf252006@yandex.ruhttps://orcid.org/0000-0001-5571-5669MasalovaOlga V.МасаловаОльга Владимировна
Russian Federation

Leading Researcher

доктор биологических наук, ведущий научный сотр., зав. лабораторией

ol.mas@mail.ruhttps://orcid.org/0000-0002-3579-4416PermyakovaKristina Yu.ПермяковаКристина Юрьевна
Russian Federation

Junior Researcher

младший научный сотр., зав. лабораторией

kristusha164@mail.ruhttps://orcid.org/0000-0003-1961-9789DemidovaNatalia A.ДемидоваНаталья Андреевна
Russian Federation

Researcher

научный сотр.

ailande@yandex.ruhttps://orcid.org/0000-0003-0365-570XValuev-EllistonVladimir T.Валуев-ЭллистонВладимир Треворович
Russian Federation

PhD, Researcher

кандидат биологических наук, научный сотрудник

gansfaust@mail.ruhttps://orcid.org/0000-0002-5659-96795776-5496IvanovAlexandr V.ИвановАлександр Владимирович
Russian Federation

Dr. Sci. Biol., Chief Researcher, head of the laboratory of biochemistry of viral infections; deputy director for scientific work

доктор биологических наук, кандидат химических наук, главный научный сотрудник, рук. лаборатории биохимии вирусных инфекций, зам. директора по научной работе

aivanov@yandex.ruhttps://orcid.org/0000-0002-3396-55336964-1715KushchAlla A.КущАлла Александровна
Russian Federation

Dr. Sci. (Biology), Professor, Leading Researcher

доктор биол. наук, проф., ведущий научный сотр.

vitallku@mail.ruGamaleya NRC of Epidemiology and Microbiology, Ministry of Health of the Russian FederationФГБУ «Национальный исследовательский центр эпидемиологии и микробиологии имени Н.Ф. Гамалеи» Минздрава РоссииMoscow State Academy of Veterinary Medicine and Biotechnology – MVA by K.I. SkryabinФГБОУ ВО «Московская государственная академия ветеринарной медицины и биотехнологии – МВА имени К.И. Скрябина» Министерства сельского хозяйства Российской ФедерацииEngelhardt Institute of Molecular Biology, Russian Academy of SciencesФГБУН «Институт молекулярной биологии имени В.А. Энгельгардта» Российской академии наук2109202368431532628062023Copyright ©; 2023, Lesnova E.I., Masalova O.V., Permyakova K.Y., Demidova N.A., Valuev-Elliston V.T., Ivanov A.V., Kushch A.A.Copyright ©; 2023, Леснова Е.И., Масалова О.В., Пермякова К.Ю., Демидова Н.А., Валуев-Эллистон В.Т., Иванов А.В., Кущ А.А.2023Lesnova E.I., Masalova O.V., Permyakova K.Y., Demidova N.A., Valuev-Elliston V.T., Ivanov A.V., Kushch A.A.Леснова Е.И., Масалова О.В., Пермякова К.Ю., Демидова Н.А., Валуев-Эллистон В.Т., Иванов А.В., Кущ А.А.https://creativecommons.org/licenses/by/4.0https://virusjour.crie.ru/jour/article/view/13159

Introduction. Hepatitis C is a liver disease with high chronicity, the cause of cirrhosis and hepatocarcinoma. The main obstacle to controlling hepatitis C is the lack of vaccines.

The aim of the work was to compare the immunogenic activity of nonstructural recombinant proteins NS3, NS4 and NS5B of hepatitis C virus (HCV) as components of a subunit candidate vaccine and to analyze the adjuvant properties of two available commercial drugs, polymuramil and pyrogenalum.

Materials and methods. BALB/c, DBA/2J and C57BL/6 mice were immunized with nonstructural proteins without adjuvants or with polymuramyl (NOD1 and NOD2 agonist) and pyrogenalum (TLR-4 agonist). The activity of antibodies was determined in ELISA, the cellular response – by antigen-specific lymphocyte proliferation and by production of IFN-γ in vitro.

Results. Recombinant proteins showed different immunogenicity. NS4 induced antibodies more efficiently than NS3 and NS5B. Significant differences were found in the immune response of three inbred lines mice: the level of IFN-γ in BALB/c and DBA/2J mice induced by NS5B protein was 30 times higher than in C57Bl/6 mice. In contrast, the induction of antibodies in BALB/c mice was lower than in C57Bl/6 and DBA/2J. Polymuramil did not increase the humoral response to NS5B and enhanced the cellular response only in C57BL/6 mice. The combined use of polymuramil with pyrogenalum significantly increased both the humoral and cellular response of mice to all recombinant HCV proteins.

Conclusion. Different immunogenic properties and different functions of recombinant non-structural HCV proteins indicate the feasibility of their combined inclusion in subunit vaccines. It was established for the first time that immunization with HCV proteins with a complex adjuvant (polymuramyl + pyrogenalum) has a synergistic effect, significantly exceeding the effect of each of them separately.

Введение. Гепатит С – заболевание печени с высокой хронизацией, являющееся причиной цирроза и гепатокарциномы. Главное препятствие для контроля за гепатитом С – отсутствие вакцин.

Цель работы – сравнение иммуногенной активности неструктурных рекомбинантных белков NS3, NS4 и NS5B вируса гепатита С (ВГС) как компонентов субъединичной кандидатной вакцины и анализ адъювантных свойств двух отечественных коммерческих препаратов: полимурамила и пирогенала.

Материалы и методы. Мышей линий BALB/c, DBA/2J и C57BL/6 иммунизировали трижды с интервалом 2 недели белками NS3, NS4 и NS5B без адъювантов или с полимурамилом (агонист рецепторов иммунного ответа NOD1 и NOD2) и пирогеналом (липополисахарид, агонист TLR-4). Гуморальный ответ определяли по активности антител в ИФА, клеточный – по индексу стимуляции пролиферации лимфоцитов и способности продуцировать IFN-γ при антигенспецифической стимуляции in vitro.

Результаты. Рекомбинантные белки проявляли различную иммуногенность. NS4 индуцировал антитела эффективнее, чем NS3 и NS5B. Значительные различия обнаружены в иммунном ответе мышей трех инбредных линий: уровень секреции IFN-γ у мышей BALB/c и DBA/2J на белок NS5B был в 30 раз выше, чем у мышей C57Bl/6. Индукция антител, напротив, у мышей BALB/c была ниже, чем у C57Bl/6 и у DBA/2J. Полимурамил не увеличивал гуморальный ответ на NS5B и усиливал клеточный ответ только у мышей C57BL/6. Сочетанное применение полимурамила с пирогеналом значительно увеличивало как гуморальный, так и клеточный ответ мышей на все рекомбинантные белки ВГС.

Выводы. Различные иммуногенные свойства и разные функции неструктурных белков ВГС в репликации вируса свидетельствуют о целесообразности их сочетанного включения в состав субъединичных вакцин. Впервые установлено, что иммунизация белками ВГС с комплексным адъювантом (полимурамил + пирогенал) оказывает синергетический эффект, значительно превышая действие каждого из них по отдельности.

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