jofinЖурнал инфектологииJournal Infectology2072-6732IPO “АIDSSPbR"10.22625/2072-6732-2017-9-4-43-52jofin-655Research ArticleОригинальные исследованияOriginal ResearchЭФФЕКТИВНОСТЬ КРОСС-ПРОТЕКТИВНОЙ РЕКОМБИНАНТНОЙ ПРОТИВОГРИППОЗНОЙ ВАКЦИНЫ, ВКЛЮЧАЮЩЕЙ КОНСЕРВАТИВНЫЕ ЭПИТОПЫ ВИРУСНЫХ БЕЛКОВ М2 И ГЕМАГГЛЮТИНИНАEFFECTIVENESS OF CROSS-PROTECTIVE RECOMBINANT INFLUENZA VACCINE BASED ON CONSERVED EPITOPES OF VIRAL PROTEINS М2 AND HEMAGGLUTININСтепановаЛ. А.StepanovaL. A.

Степанова Людмила Алексеевна – ведущий научный сотрудник,  кандидат биологических наук.

 

Saint-Petersburg.

stepanoval60@mail.ruШуклинаМ. А.ShuklinaM. A.

Шуклина Марина Александровна – младший научный сотрудник.

 

Saint-Petersburg.

jigokutsushin@mail.ruБлохинаЕ. А.BlokhinaE. A.

Блохина Елена Александровна – научный сотрудник, кандидат биологических наук. Москва.

Moscow.

blohina-lena87@mail.ruКотляровР. Ю.KotlyarovR. Y.

Котляров Роман Юрьевич – научный сотрудник, кандидат биологических наук.

Москва.

Moscow.

kotlyarov@biengi.ac.ruКовалеваА. А.KovalevaA. A.

Ковалева Анна Александровна – научный сотрудник.

Санкт-Петербург.

Saint-Petersburg.

gifer@rambler.ruРавинН. В.RavinN. V.

Равин Николай Викторович – заместитель директора по научной работе, доктор биологических наук.

Москва.

Moscow.

nravin@biengi.ac.ruЦыбаловаЛ. М.TsybalovaL. M.

Цыбалова Людмила Марковна – заместитель директора по научной работе, заведующая отделом вакцинологии, доктор медицинских наук.

Санкт-Петербург.

Saint-Petersburg.

sovet@influenza.spb.ruНаучно-исследовательский институт гриппа.РоссияScience Research Institute of Influenza.Russian FederationФедеральный исследовательский центр «Фундаментальные основы биотехнологии».РоссияFederal Research Centre «Fundamentals of Biotechnology».Russian Federation201709012018944352Copyright © Степанова Л.А., Шуклина М.А., Блохина Е.А., Котляров Р.Ю., Ковалева А.А., Равин Н.В., Цыбалова Л.М., 20182018Степанова Л.А., Шуклина М.А., Блохина Е.А., Котляров Р.Ю., Ковалева А.А., Равин Н.В., Цыбалова Л.М.Stepanova L.A., Shuklina M.A., Blokhina E.A., Kotlyarov R.Y., Kovaleva A.A., Ravin N.V., Tsybalova L.M.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.niidi.ru/jofin/article/view/655

Вирус гриппа является наиболее уникальным по уровню изменчивости антигенных и биологических свойств. Из-за постоянных мутаций в генах, кодирующих поверхностные белки вируса, в существующих «сезонных» вакцинах приходится ежегодно заменять 1–2 вирусных компонента. Кроме того, традиционные вакцины являются штамм-специфическими и обладают ограниченной эффективностью в предотвращении заболеваний, вызванных новыми штаммами вирусов гриппа. В связи с этим создание противогриппозных вакцин на основе консервативных детерминант вирусных белков с широким спектром защиты и коротким периодом производства является одной из приоритетных задач, решение которой приведет к реальному контролю гриппозной инфекции. Перспективной тенденцией в создании универсальных гриппозных вакцин является конструирование рекомбинантных белков на основе комбинации консервативных вирусных белков или пептидов. Цель: разработка кандидатной рекомбинантной гриппозной вакцины на основе двух консервативных белков вируса гриппа А (М2 и НА) и оценка ее иммуногенности и защитного эффекта на животной модели. Результаты: исследовали гуморальный и Т-клеточный ответ у мышей (Balb/c) после интраназальной иммунизации рекомбинантными белками (Flg4M2ehs и Flg-HA2-2-4M2ehs). Оба белка стимулировали формирование выраженного М2е-специфического гуморального и CD4+ Т-клеточного ответа в легких мышей. Рекомбинантный белок с двумя таргетными антигенами (М2е и полипептид 76–130 второй субъединицы гемагглютинина) стимулирует формирование вирус-специфического Т-клеточного ответа и полную защиту (100% выживаемость) мышей от летального заражения вирусами гриппа А человека и птиц (A/H3N2, A/H2N2, A/H5N1). У мышей, иммунизированных рекомбинантным белком с одним таргетным антигеном (Flg4M2ehs), выживаемость после летального заражения составила 60–75%. Заключение: полученные результаты демонстрируют существенную роль консервативного фрагмента второй субъединицы гемагглютинина в формировании протективного Т-клеточного иммунитета и защите мышей от летального заражения вирусами гриппа А различных субтипов. Показана перспективность разработки вакцинного препарата на основе двух консервативных антигенных детерминант вирусов гриппа А. 

The influenza virus is the most unique in the level of variability of antigenic and biological properties. Because of constant mutations into genes coding surface viral proteins, in modern vaccines it is necessary to replace 1–2 virus components annually. Traditional influenza vaccines are the strain – specific and have limited efficiency in prevention of new strains of influenza viruses. In this regard, creation of influenza vaccines based on conserved determinants of viral proteins with broad spectrum protection and the short period of production is one of priority tasks which decision will lead to real control of an influenza infection. A current trend in the design of universal flu vaccines is the construction of recombinant proteins based the combination of conserved viral proteins or peptides. The goals of this study: to develop the candidate recombinant flu vaccine based on the two conserved influenza proteins (М2 and НА); to investigate immune response; and to measure the protection activity in an animal model. Results: In this study we investigated the humoral and T-cell response in mice after intranasal immunization with recombinant proteins (Flg-4M2ehs and Flg-HA2-2-4M2ehs). Both proteins induce a robust M2e-specific humoral and CD4+ T-cell response in mice lung. The recombinant protein with two target antigens (M2e and HA2) induces virusspecific CD4+ and CD8+ T-cell response and full protection (100% survival) of mice from lethal challenge human and avian influenza viruses A (A/H3N2, A/H2N2, A/H5N1). In mice immunized with Flg-4M2ehs, the survival after lethal challenge was 60–75%. Conclusion: Our results show an essential role of a conserved fragment of the HA2 in the formation of protective T-cell response and protection of mice from lethal challenge with influenza viruses A of various subtypes. The prospects of the development of vaccine formulation based on two conserved antigenic determinants of influenza virus A are shown.

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