Acta NaturaeActa NaturaeActa Naturae2075-8251Acta Naturae Ltd2777410.32607/actanaturae.27774Research ArticlesЭкспериментальные статьиResearch ArticleAdministration of rAAV encoding monoclonal antibodies protects mice challenged with a lethal dose of H3N2 influenza virus and neutralizes H1 and H3 strainsВведение rAAV, кодирующих широко нейтрализующие антитела, обеспечивает защиту мышей при заражении летальной дозой вируса гриппа H3N2 и нейтрализацию штаммов подтипов H1 и H3MintaevR. R.МинтаевР. Р.
Russian Federation
ramil.mintaev@fbb.msu.ruUrusovF. A.УрусовФ. А.
Russian Federation
ramil.mintaev@fbb.msu.ruSolovievA. V.СоловьевА. В.
Russian Federation
ramil.mintaev@fbb.msu.ruBeluginB. V.БелугинБ. В.
Russian Federation
ramil.mintaev@fbb.msu.ruGlazkovaD. V.ГлазковаД. В.
Russian Federation
ramil.mintaev@fbb.msu.ruShipulinG. A.ШипулинГ. А.
Russian Federation
ramil.mintaev@fbb.msu.ruBogoslovskayaE. V.БогословскаяЕ. В.
Russian Federation
ramil.mintaev@fbb.msu.ruFederal State Budgetary Institution “Center for Strategic Planning and Management of Medical and Biological Health Risks”, Federal Medical-Biological AgencyФедеральное государственное бюджетное учреждение «Центр стратегического планирования и управления медико-биологическими рисками здоровью» Федерального медико-биологического агентства2204202618164711008202518092025Copyright ©; 2026, Mintaev R.R., Urusov F.A., Soloviev A.V., Belugin B.V., Glazkova D.V., Shipulin G.A., Bogoslovskaya E.V.Copyright ©; 2026, Минтаев Р.Р., Урусов Ф.А., Соловьев А.В., Белугин Б.В., Глазкова Д.В., Шипулин Г.А., Богословская Е.В.2026Mintaev R.R., Urusov F.A., Soloviev A.V., Belugin B.V., Glazkova D.V., Shipulin G.A., Bogoslovskaya E.V.Минтаев Р.Р., Урусов Ф.А., Соловьев А.В., Белугин Б.В., Глазкова Д.В., Шипулин Г.А., Богословская Е.В.https://creativecommons.org/licenses/by/4.0https://actanaturae.ru/2075-8251/article/view/27774

The influenza virus causes seasonal epidemics throughout the world. At the same time, the rapid mutation of the virus renders the use of seasonal vaccines less effective. One of the approaches sought to improve influenza prevention is the use of monoclonal antibodies that are active against a wide range of influenza virus strains. In this study, the virus-neutralizing activity of the monoclonal antibodies CR9114, MHAA4549A, MEDI8852, C585, and 1G01 against the influenza virus was assessed. To this end, recombinant vectors based on adeno-associated virus (rAAV) encoding these antibodies were used. The rAAV vectors were expressed in mice in vivo, and the virus-neutralizing activity of the sera against the H1N1 and H3N2 influenza virus strains was assessed. Administration of rAAV-C585, rAAV-MHAA4549A, and rAAV-1G01 conferred 100% protection to mice challenged with a lethal dose of the H3N2 influenza virus. The efficacy of rAAV-CR9114 and rAAV-MEDI8852 against this influenza virus strain was lower, at 80 and 75%, respectively.

Вирус гриппа вызывает сезонные эпидемии во всем мире. Вместе с тем, быстрая эволюция вируса делает применение сезонных вакцин недостаточно эффективным. Одним из подходов к решению проблемы профилактики гриппа может быть использование моноклональных антител, активных в отношении широкого спектра штаммов вируса гриппа. В представленной работе определяли вируснейтрализующую активность моноклональных антител CR9114, MHAA4549A, MEDI8852, С585 и 1G01 против вируса гриппа. С этой целью использовали рекомбинантные векторы на основе аденоассоциированного вируса (rAAV), кодирующие эти антитела. rAAV-векторы экспрессировали в мышах in vivo и определяли вируснейтрализующую активность сывороток по отношению к штаммам H1N1 и H3N2 вируса гриппа. Введение rAAV-С585, rAAV-MHAA4549A и rAAV-1G01 обеспечивало 100% защиту мышей, зараженных летальной дозой вируса гриппа H3N2. Эффективность rAAV-CR9114 и rAAV-MEDI8852 против данного штамма вируса гриппа оказалась ниже – 80 и 75% соответственно.

rAAVmonoclonal antibodiesCR9114MHAA4549AMEDI8852C5851G01influenzarAAVмоноклональные антителаCR9114MHAA4549AMEDI8852С5851G01гриппGovernment of the Russian FederationПравительство Российской ФедерацииThis work was completed as part of a state assignment of the Federal Medical and Biological Agency.Работа выполнена в рамках государственного задания ФМБА.Iuliano AD, Roguski KM, Chang HH, et al. Estimates of global seasonal influenza-associated respiratory mortality: a modelling study. Lancet. 2018;391:10127(1285-1300). doi: 10.1016/S0140-6736(17)33293-2CDC seasonal flu vaccine effectiveness studies. Centers for Disease Control and Prevention. Updated May 30, 2025. Accessed July 15, 2025. https://www.cdc.gov/flu/vaccines-work/effectiveness-studies.htmCDC.gov [Internet]. CDC Seasonal Flu Vaccine Effectiveness Studies, Inc; 2009-2025 [updated 2025 May 30]. Available from: https://www.cdc.gov/flu/vaccines-work/effectiveness-studies.htmDimmock NJ. Neutralization of animal viruses. Curr Top Microbiol Immunol. 1993;183:1-149. doi: 10.1007/978-3-642-77849-0Pelegrin M, Naranjo-Gomez M, Piechaczyk M. Antiviral Monoclonal Antibodies: Can They Be More Than Simple Neutralizing Agents? Trends Microbiol. 2015;23(10):653-665. doi: 10.1016/j.tim.2015.07.005Sun X, Ling Z, Yang Z, Sun B. Broad neutralizing antibody-based strategies to tackle Influenza. Curr Opin Virol. 2022;53:101207. doi: 10.1016/j.coviro.2022.101207Sun X, Ling Z, Yang Z, Sun B. Broad neutralizing antibody-based strategies to tackle Influenza. Curr. Opin. In Vir. 2022;53:101207. doi: 10.1016/j.coviro.2022.101207Gehring AJ, Salimzadeh L. Current and future use of antibody-based passive immunity to prevent or control HBV/HDV infections. Antiviral Res. 2024;226:105893. doi: 10.1016/j.antiviral.2024.105893Gehring AJ, Salimzadeh L. Current and future use of antibody-based passive immunity to prevent or control HBV/HDV infections. Antiviral Research. 2024;226:105893. doi: 10.1016/j.antiviral.2024.105893Chavda VP, Prajapati R, Lathigara D, et al. Therapeutic monoclonal antibodies for COVID-19 management: an update. Expert Opin Biol Ther. 2022;22(6):763-780. doi: 10.1080/14712598.2022.2078160Chavda VP, Prajapati R, Lathigara D, Nagar B, Kukadiya J, Redwan EM, Uversky VN, Kher MN, Patel R. Therapeutic monoclonal antibodies for COVID-19 management: an update. Expert Opinion on Biological Therapy. 2022;22(6):763-780. doi: 10.1080/14712598.2022.2078160Li L, Zhang W, Hu Y, et al. Effect of Convalescent Plasma Therapy on Time to Clinical Improvement in Patients With Severe and Life-threatening COVID-19: A Randomized Clinical Trial. JAMA. 2020;324(5):460-470. doi: 10.1001/jama.2020.10044Li L, Zhang W, Hu Y, Tong X, et al. Effect of Convalescent Plasma Therapy on Time to Clinical Improvement in Patients With Severe and Life-threatening COVID-19: A Randomized Clinical Trial. JAMA. 2020;324(5):460-470. doi: 10.1001/jama.2020.10044Perotti C, Baldanti F, Bruno R, et al. Mortality reduction in 46 severe Covid-19 patients treated with hyperimmune plasma. A proof of concept single arm multicenter trial. Haematologica. 2020;105(12):2834-2840. doi: 10.3324/haematol.2020.261784Perotti C, Baldanti F, Bruno R, Del Fante C, et al. Covid-Plasma Task Force. Mortality reduction in 46 severe Covid-19 patients treated with hyperimmune plasma. A proof of concept single arm multicenter trial. Haematologica. 2020;105(12):2834-2840. doi: 10.3324/haematol.2020.261784Ali SO, Takas T, Nyborg A, et al. Evaluation of MEDI8852, an Anti-Influenza A Monoclonal Antibody, in Treating Acute Uncomplicated Influenza. Antimicrob Agents Chemother. 2018;62(11):e00694-18. doi: 10.1128/aac.00694-18Ali SO, Takas T, Nyborg A, Shoemaker K, Kallewaard NL, et al. Evaluation of MEDI8852, an Anti-Influenza A Monoclonal Antibody, in Treating Acute Uncomplicated Influenza. Antimicrob Agents Chemother. 2018;62(11):e00694-18. doi: 10.1128/aac.00694-18McBride JM, Lim JJ, Burgess T, et al. Phase 2 Randomized Trial of the Safety and Efficacy of MHAA4549A, a Broadly Neutralizing Monoclonal Antibody, in a Human Influenza A Virus Challenge Model. Antimicrob Agents Chemother. 2017;61(11):e01154-17. doi: 10.1128/AAC.01154-17McBride JM, Lim JJ, Burgess T, Deng R, et al. Phase 2 Randomized Trial of the Safety and Efficacy of MHAA4549A, a Broadly Neutralizing Monoclonal Antibody, in a Human Influenza A Virus Challenge Model. Antimicrob Agents Chemother. 2017;61(11):e01154-17. doi: 10.1128/AAC.01154-17. PMID: 28807912; PMCID: PMC5655070.Lim JJ, Nilsson AC, Silverman M, et al. A Phase 2 Randomized, Double-Blind, Placebo-Controlled Trial of MHAA4549A, a Monoclonal Antibody, plus Oseltamivir in Patients Hospitalized with Severe Influenza A Virus Infection. Antimicrob Agents Chemother. 2020;64(7):e00352-20. doi: 10.1128/AAC.00352-20Lim JJ, Nilsson AC, Silverman M, Assy N, et al. A Phase 2 Randomized, Double-Blind, Placebo-Controlled Trial of MHAA4549A, a Monoclonal Antibody, plus Oseltamivir in Patients Hospitalized with Severe Influenza A Virus Infection. Antimicrob Agents Chemother. 2020;64(7):e00352-20. doi: 10.1128/AAC.00352-20. PMID: 32393496; PMCID: PMC7318030.Qiu X, Audet J, Wong G, et al. Successful treatment of ebola virus-infected cynomolgus macaques with monoclonal antibodies. Sci Transl Med. 2012;4(138):138ra81. doi: 10.1126/scitranslmed.3003876Qiu X, Audet J, Wong G, Pillet S, Bello A, Cabral T, Strong JE, Plummer F, Corbett CR, Alimonti JB, Kobinger GP. Successful treatment of ebola virus-infected cynomolgus macaques with monoclonal antibodies. Sci Transl Med. 2012;4(138):138ra81. doi: 10.1126/scitranslmed.3003876Limberis MP, Tretiakova A, Nambiar K, et al. Adeno-Associated Virus Serotype 9-Expressed ZMapp in Mice Confers Protection Against Systemic and Airway-Acquired Ebola Virus Infection. J Infect Dis. 2016;214(12):1975-1979. doi: 10.1093/infdis/jiw460Limberis MP, Tretiakova A, Nambiar K, Wong G, Racine T, Crosariol M, Xiangguo Q, Kobinger G, Wilson JM. Adeno-Associated Virus Serotype 9-Expressed ZMapp in Mice Confers Protection Against Systemic and Airway-Acquired Ebola Virus Infection. J Infect Dis. 2016;214(12):1975-1979. doi: 10.1093/infdis/jiw460Martinez-Navio JM, Fuchs SP, Pantry SN, et al. Adeno-Associated Virus Delivery of Anti-HIV Monoclonal Antibodies Can Drive Long-Term Virologic Suppression. Immunity. 2019;50(3):567-575.e5. doi: 10.1016/j.immuni.2019.02.005Martinez-Navio JM, Fuchs SP, Pantry SN, Lauer WA, Duggan NN, Keele BF, Rakasz EG, Gao G, Lifson JD, Desrosiers RC. Adeno-Associated Virus Delivery of Anti-HIV Monoclonal Antibodies Can Drive Long-Term Virologic Suppression. Immunity. 2019;50(3):567-575.e5. doi: 10.1016/j.immuni.2019.02.005Leontyev DS, Urusov FA, Glazkova DV, et al. Study of the protective efficacy of CombiMab-2 against human immunodeficiency virus type 1 in mice humanised with CD4+ T-lymphocytes. Biological Products. Prevention, Diagnosis, Treatment. 2024;24(3):312-321. doi: 10.30895/2221-996X-2024-24-3-312-321Леонтьев ДС, Урусов ФА, Глазкова ДВ, Белугин БВ, Орлова ОВ, Минтаев РР, Цыганова ГМ, Богословская ЕВ, Шипулин ГА. Исследование защитной эффективности препарата КомбиМаб-2 против вируса иммунодефицита человека типа 1 на мышах, гуманизированных CD4+ Т-лимфоцитами. БИОпрепараты. Профилактика, диагностика, лечение. 2024;24(3):312-321. doi: 10.30895/2221-996X-2024-24-3-312-321Adam VS, Crosariol M, Kumar S, et al. Adeno-associated virus 9-mediated airway expression of antibody protects old and immunodeficient mice against influenza virus. Clin Vaccine Immunol. 2014;21(11):1528-1533. doi: 10.1128/CVI.00572-14Adam VS, Crosariol M, Kumar S, Ge MQ, Czack SE, Roy S, Haczku A, Tretiakova A, Wilson JM, Limberis MP. Adeno-associated virus 9-mediated airway expression of antibody protects old and immunodeficient mice against influenza virus. Clin Vaccine Immunol. 2014;21(11):1528-1533.Del Rosario JMM, Smith M, Zaki K, et al. Protection From Influenza by Intramuscular Gene Vector Delivery of a Broadly Neutralizing Nanobody Does Not Depend on Antibody Dependent Cellular Cytotoxicity. Front Immunol. 2020;11:627. doi: 10.3389/fimmu.2020.00627Del Rosario JMM, Smith M, Zaki K, Risley P, Temperton N, Engelhardt OG, Collins M, Takeuchi Y, Hufton SE. Protection From Influenza by Intramuscular Gene Vector Delivery of a Broadly Neutralizing Nanobody Does Not Depend on Antibody Dependent Cellular Cytotoxicity. Front Immunol. 2020;11:627. doi: 10.3389/fimmu.2020.00627Dreyfus C, Laursen NS, Kwaks T, et al. Highly conserved protective epitopes on influenza B viruses. Science. 2012;337(6100):1343-1348. doi: 10.1126/science.1222908Dreyfus C, Laursen NS, Kwaks T, Zuijdgeest D, et al. Highly conserved protective epitopes on influenza B viruses. Science. 2012;337(6100):1343-1348. doi: 10.1126/science.1222908Nakamura G, Chai N, Park S, et al. An in vivo human-plasmablast enrichment technique allows rapid identification of therapeutic influenza A antibodies. Cell Host Microbe. 2013;14(1):93-103. doi: 10.1016/j.chom.2013.06.004Nakamura G, Chai N, Park S, Chiang N, et al. An in vivo human-plasmablast enrichment technique allows rapid identification of therapeutic influenza A antibodies. Cell Host Microbe. 2013;14(1):93-103. doi: 10.1016/j.chom.2013.06.004Kallewaard NL, Corti D, Collins PJ, et al. Structure and Function Analysis of an Antibody Recognizing All Influenza A Subtypes. Cell. 2016;166(3):596-608. doi: 10.1016/j.cell.2016.05.073Kallewaard NL, Corti D, Collins PJ, Neu U, et al. Structure and Function Analysis of an Antibody Recognizing All Influenza A Subtypes. Cell. 2016;166(3):596-608. doi: 10.1016/j.cell.2016.05.073Qiu Y, Stegalkina S, Zhang J, et al. Mapping of a novel H3-specific broadly neutralizing monoclonal antibody targeting the hemagglutinin globular head isolated from an elite influenza virus-immunized donor exhibiting serological breadth. J Virol. 2020;94(6):e01035-19. doi: 10.1128/JVI.01035-19Qiu Y, Stegalkina S, Zhang J, Boudanova E, et al. Mapping of a novel H3-specific broadly neutralizing monoclonal antibody targeting the hemagglutinin globular head isolated from an elite influenza virus-immunized donor exhibiting serological breadth. J Virol. 2020;94:e01035-19.Stadlbauer D, Zhu X, McMahon M, et al. Broadly protective human antibodies that target the active site of influenza virus neuraminidase. Science. 2019;366(6464):499-504. doi: 10.1126/science.aay0678Stadlbauer D, Zhu X, McMahon M, Turner JS, et al. Broadly protective human antibodies that target the active site of influenza virus neuraminidase. Science. 2019;366(6464):499-504. doi: 10.1126/science.aay0678Paules CI, Lakdawala S, McAuliffe JM, et al. The Hemagglutinin A Stem Antibody MEDI8852 Prevents and Control Disease and Limits Transmission of Pandemic Influenza Viruses. J Infect Dis. 2017;216(3):356-365. doi: 10.1093/infdis/jix292Paules CI, Lakdawala S, McAuliffe JM, Paskel M, Vogel L, Kallewaard NL, Zhu Q, Subbarao K. The Hemagglutinin A Stem Antibody MEDI8852 Prevents and Control Disease and Limits Transmission of Pandemic Influenza Viruses. J Infect Dis. 2017;216(3):356-365. doi: 10.1093/infdis/jix292Shipulin GA, Glazkova DV, Urusov FA, et al. Triple Combinations of AAV9-Vectors Encoding Anti-HIV bNAbs Provide Long-Term In Vivo Expression of Human IgG Effectively Neutralizing Pseudoviruses from HIV-1 Global Panel. Viruses. 2024;16(8):1296. doi: 10.3390/v16081296Shipulin GA, Glazkova DV, Urusov FA, Belugin BV, Dontsova V, Panova AV, Borisova AA, Tsyganova GM, Bogoslovskaya EV. Triple Combinations of AAV9-Vectors Encoding Anti-HIV bNAbs Provide Long-Term In Vivo Expression of Human IgG Effectively Neutralizing Pseudoviruses from HIV-1 Global Panel. Viruses. 2024;16(8):1296. doi: 10.3390/v16081296Cottey R, Rowe CA, Bender BS. Influenza virus. Curr Protoc Immunol. 2001;42(1): 19.1.1-19.1.32. doi: 10.1002/0471142735.im1911s42Cottey R, Rowe CA, Bender BS. Influenza virus. Current protocols in immunology. 2001;42(1):19-1. doi: 10.1002/0471142735.im1911s42Reed LJ, Muench H. A simple method of estimation fifty per cent endpoints. Am J Hyg. 1938;27(3):493-497.Reed LJ, Muench H. A simple method of estimation of 50% end points. American J Hyg. 1938;27:493-497.Zost SJ, Gilchuk P, Case JB, et al. Potently neutralizing and protective human antibodies against SARS-CoV-2. Nature. 2020;584(7821):443-449. doi: 10.1038/s41586-020-2548-6Zost SJ, Gilchuk P, Case JB, Binshtein E, et al. Potently neutralizing and protective human antibodies against SARS-CoV-2. Nature. 2020;584(7821):443-449. doi: 10.1038/s41586-020-2548-6Sedeyn K, Saelens X. New antibody-based prevention and treatment options for influenza. Antiviral Res. 2019;170:104562. doi: 10.1016/j.antiviral.2019.104562Sedeyn K, Saelens X. New antibody-based prevention and treatment options for influenza. Antiviral Research. 2019;170:104562. doi: 10.1016/j.antiviral.2019.104562Jang YH, Seong BL. The Quest for a Truly Universal Influenza Vaccine. Front Cell Infect Microbiol. 2019;9:344. doi: 10.3389/fcimb.2019.00344Beukenhorst AL, Frallicciardi J, Koch CM, et al. The influenza hemagglutinin stem antibody CR9114: evidence for a narrow evolutionary path towards universal protection. Front Virol. 2022;2:1049134. doi: 10.3389/fviro.2022.1049134Beukenhorst AL, Frallicciardi J, Koch CM, Klap JM, Phillips A, et al. The influenza hemagglutinin stem antibody CR9114: Evidence for a narrow evolutionary path towards universal protection. Frontiers in Virology. 2022;2:1049134. doi: 10.3389/fviro.2022.1049134Sutton TC, Lamirande EW, Bock KW, et al. In Vitro Neutralization Is Not Predictive of Prophylactic Efficacy of Broadly Neutralizing Monoclonal Antibodies CR6261 and CR9114 against Lethal H2 Influenza Virus Challenge in Mice. J Virol. 2017;91(24):e01603-17. doi: 10.1128/JVI.01603-17Sutton TC, Lamirande EW, Bock KW, Moore IN, Koudstaal W, Rehman M, Weverling GJ, Goudsmit J, Subbarao K. In Vitro Neutralization Is Not Predictive of Prophylactic Efficacy of Broadly Neutralizing Monoclonal Antibodies CR6261 and CR9114 against Lethal H2 Influenza Virus Challenge in Mice. J Virol. 2017;91(24):e01603-17. doi: 10.1128/JVI.01603-17. PMID: 29046448; PMCID: PMC5709608.Chai N, Swem LR, Reichelt M, et al. Two escape mechanisms of influenza A virus to a broadly neutralizing stalk-binding antibody. PLoS Pathog. 2016;12(6):e1005702. doi: 10.1371/journal.ppat.1005702Chai N, Swem LR, Reichelt M, et al. Two escape mechanisms of influenza A virus to a broadly neutralizing stalk-binding antibody. PLoS Pathog. 2016;12:e1005702. doi: 10.1371/journal.ppat.1005702Jagadisan B, Dhawan A. Hepatotoxicity in adeno-associated viral vector gene therapy. Curr Hepatol Rep. 2023;22(4):276-290. doi: 10.1007/s11901-023-00624-5Jagadisan B, Dhawan A. Hepatotoxicity in adeno-associated viral vector gene therapy. Current Hepatology Reports. 2023;22(4):276-290. doi: 10.1007/s11901-023-00624-5Duan D. Lethal immunotoxicity in high-dose systemic AAV therapy. Mol Ther. 2023;31(11):3123-3126. doi: 10.1016/j.ymthe.2023.10.015Duan D. Lethal immunotoxicity in high-dose systemic AAV therapy. Molecular Therapy. 2023;31(11):3123-3126. doi: 10.1016/j.ymthe.2023.10.015