Development of a candidate influenza vaccine based on virus-like particles displaying influenza M2e peptide into the immunodominant region of hepatitis B core antigen: Broad protective efficacy of particles carrying four copies of M2e

Цыбалова, Л. М.; Степанова, Л. А.; Kuprianov, Victor V.; Blokhina, Elena; Potapchuk, M. V.; Korotkov, Alexandr V.; Горшков, А. Н.; Kasyanenko, Marina A.; Ravin, Nikolai V.; Киселев, О. И.

doi:10.1016/j.vaccine.2015.04.073

Cited by 32 publications

(30 citation statements)

References 30 publications

(38 reference statements)

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“…Ранее мы высказали предположение [13], что рекомбинантные кросс-протективные вакцины могут быть использованы для праймирования детей. Иммунизация такой вакциной не препятствует развитию естественной инфекции в легкой форме и соответственно формированию полноценного иммунного ответа на все вирусные белки, что совершенно исключается при ежегодной вакцинации детей субъединичными вакцинами.…”

Section: Discussionunclassified

“…Препарат плацебо содержит те же компоненты, кроме белка HBс4M2e, который замещён белком HBc. Рекомбинантные белки были сконструированы и тестированы совместно со специалистами Федерального исследовательского центра биотехнологии РАН [12,13]. Белки получены путем культивирования штаммов E. coli DLT1270 pQE60HBc/4Me2h и E.сoli DLT1270 pQE60HBc соответственно и очищены с помощью гельфильтрации и ионообменной хроматографии.…”

Section: Introductionunclassified

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Cross-Protective Properties of an Influenza Vaccine Based on Hbc4m2e Recombinant Protein

Цыбалова

Степанова

Shuklina

et al. 2018

Problems of Virology

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One of the main problems in the area of influenza prophylaxis and pandemic prevention is the development of cross-reactive vaccines, i.e. vaccines directed against all subtypes of human influenza viruses. Such vaccines are being developed in many countries for more than 10 years. A number of vaccines are presently undergoing clinical trials. We created Uniflu candidate vaccine based on recombinant HBc4M2e protein consisting of 4 tandem-connected copies of the highly conserved ectodomain of M2 protein of the influenza A virus. These 4 copies were genetically fused to the carrier protein, namely hepatitis B core antigen. Commercially available Derinat was used as adjuvant in the candidate vaccine. Preclinical studies on laboratory animals (mice, ferrets) demonstrated that immunization with Uniflu leads to significantly higher level of specific immunoglobulins in the blood and bronchoalveolar lavages. Moreover, it produces immunoglobulins belonging to subtype IgG2a that is the most important mediator of antibody-dependent cytotoxicity. The vaccine under review stimulates the proliferation of T-lymphocytes, as well as the formation of CD4+ and CD8+ T-cells synthesizing ɣ-IFN. When infected with the lethal doses (5 LD50) of influenza A viruses of the subtypes H1N1, H2N2, H3N2, and H1N1pdm09, immunized animals typically developed mild form of illness. This kept them alive in 90-100% of cases, which demonstrated almost complete protection from death. Replication of the virus in the lungs of immunized mice was reduced by 1.8-4.8 log10. High immunogenicity of the vaccine, and reduced clinical symptoms following experimental infection, were demonstrated in ferrets as well. The developed recombinant vaccine Uniflu has high specific activity and cross-protection. Uniflu can be proposed as pre-pandemic vaccine, provided that it passes clinical trials.

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Section: Discussionunclassified

Section: Introductionunclassified

Cross-Protective Properties of an Influenza Vaccine Based on Hbc4m2e Recombinant Protein

Цыбалова

Степанова

Shuklina

et al. 2018

Problems of Virology

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“…Как видно на рисунке 1Б в белках FlgНА-4М2е и Flg4М2е целевые белки находятся на С-конце флагеллина. Последовательность аминокислот в пептиде М2е вируса птиц А(Н5N1), по сравнению с консенсусной последовательностью аминокислот вирусов гриппа человека отличается на 3 аминокислоты в положениях 8 (Т → I), 15 (Е → G) и 19 (S fiN); по сравнению с М2е вируса А/ Калифорния/05/09, имеет одну лишь замену N12S и индуцирует антитела, хорошо взаимодействующие с М2е вируса А (Н1N1)pdm09 [15].…”

Section: а б вunclassified

Strengthening the Effectiveness of the Candidate Influenza Vaccine by Combining Conserved Sequences of Hemagglutinin and M2 protein

Цыбалова¹,

Степанова²,

Kotlyarov³

et al. 2017

Epidemiology and Vaccine Prevention

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1 ФГБУ «НИИ гриппа» Минздрава России, Санкт-Петербург 2 ФГУ «ФИЦ «Фундаментальные основы биотехнологии» РАН, Москва Усиление эффективности кандидатной вакцины против гриппа сочетанием консервативных последовательностей гемагглютинина и М2 белка Резюме Разработка универсальной вакцины против гриппа -вакцины, направленной на все субтипы вирусов гриппа человека, -является наиболее актуальной современной задачей в специфической профилактике гриппа. В настоящей статье дана сравнительная характеристика специфической активности нескольких вариантов рекомбинантного вакцинного белка, включающего антигенные детерминанты вируса гриппа -эктодомен белка М2 (M2е) и фрагмент второй субъединицы гемагглютинина (аминокислотная последовательность 76 -130). В качестве белка-носителя и адъюванта был выбран флагеллин -белок Salmonella typhimurium как полноразмерный, так и с делетированной гипервариабельной частью. На экспериментальных животных показана высокая иммуногенность препаратов и способность предотвращать летальность при инфицированнии вирусом гриппа. Наибольшей протективностью обладал гибридный Abstract (the amino acid sequence 76 -130). Flagellin -Salmonella typhimurium protein was used as carrier protein and as adjuvant. We use two forms of flagellin: full size and with deleted hypervariable region. The proteins showed high immunogenicity, and the ability to prevent lethal infection of influenza virus in mice. and M2e on the C-terminus (protein Flg-HA2-4M2e) demonstrated the most protective properties. It provides 100% survival immunized mice that were challenge with a high dose of influenza A (H3N2) -10 LD50. Proteins containing only full sized flagellin with M2e or flagellin truncated form with M2e at the C-terminus and HA2 within the hypervariable region, protected 75% of animals from lethal infection. Protein Flg-HA2-4M2e is promising for further study as a vaccine. The development of universal influenza vaccine -a vaccine directed to all subtypes of human influenza A viruses -is the really actual problem task. This paper presents the comparative characteristic of the specific activity of various recombinant proteins consisting of antigenic determinants of influenza A virus -the ectodomain of the M2 protein (M2e) and a fragment of the second subunit of the hemagglutinin

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“…Ранее нами была показана возможность встраивания до 4 копий М2е-пептида в район иммунодоминантной петли HBc [16]. Такие ВПЧ обладали высокой иммуногенностью и обеспечивали защиту иммунизированных животных от заражения различными штаммами вируса гриппа А [17].…”

Section: Introductionunclassified

Construction of Mosaic HBC Particles Presenting Conservative Fragments of M2 Protein and Hemagglutinin of Influenza a Virus

Blokhina

Ravin

2018

Problems of Virology

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Virus-like HBc particles formed as a result of the self-assembly of the nuclear antigen of the hepatitis B virus can be used as a highly immunogenic carrier for the presentation of foreign epitopes when creating recombinant vaccines. We use this vehicle to create influenza vaccines based on the conservative antigens of the influenza virus, the extracellular domain of the transmembrane protein M2 (M2e) and the fragment of the second subunit of hemagglutinin (HA2). Presentation on the surface of HBc particles should improve the immunogenicity of these peptides. Using genetic engineering techniques, we obtained a fusion protein in which the HA2 sequence is attached to the N-terminus of the HBc antigen, and the M2e peptide is included in the immunodominant loop region exposed on the surface of HBc particle. The hybrid protein expressed in Escherichia coli and purified under denaturing conditions formed virus-like HBc particles after refolding in vitro. Refolding of this protein in the presence of a previously denatured HBc antigen carrying no inserts resulted in formation of mosaic virus-like particles. The developed method will allow construction of mosaic HBc particles carrying different target epitopes of the influenza virus by combining the corresponding modified HBc proteins, which opens the possibility of creating vaccines with a wider spectrum of protection.

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Development of a candidate influenza vaccine based on virus-like particles displaying influenza M2e peptide into the immunodominant region of hepatitis B core antigen: Broad protective efficacy of particles carrying four copies of M2e

Cited by 32 publications

References 30 publications

Cross-Protective Properties of an Influenza Vaccine Based on Hbc4m2e Recombinant Protein

Cross-Protective Properties of an Influenza Vaccine Based on Hbc4m2e Recombinant Protein

Strengthening the Effectiveness of the Candidate Influenza Vaccine by Combining Conserved Sequences of Hemagglutinin and M2 protein

Construction of Mosaic HBC Particles Presenting Conservative Fragments of M2 Protein and Hemagglutinin of Influenza a Virus

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