Abstract
Highly pathogenic H5Nx avian influenza viruses constantly threaten the poultry industry and humans and have pandemic potential. These viruses continuously evolve, requiring a universal vaccine to protect chickens from members of diverse clades. The purpose of this study was to develop an H5 cleavage-site peptide vaccine containing polybasic amino acids (RRRK) to completely protect chickens from H5N6, H5N8, and H5N1 avian influenza viruses. Chickens were immunized with various doses of a keyhole limpet hemocyanin (KLH)-conjugated H5 cleavage-site peptide vaccine containing RRRK. The effect of RRRK was evaluated by comparing the survival rates of chickens immunized with vaccines either containing or lacking RRRK. The ability of the RRRK-containing vaccine to confer long-term protective immunity was also assessed. We found that protection was dependent on the number of antigens in the vaccine containing RRRK. Chickens immunized intramuscularly with two doses of 5 μg of the vaccine containing RRRK were completely protected, but those immunized with fewer than two doses of 3 or 1 μg were not protected. Chickens immunized with the vaccine lacking RRRK were not protected, suggesting the importance of the polybasic amino acids in conferring immunity. Our results suggest that conserved H5 cleavage-site peptides with polybasic amino acids may be a potential universal vaccine to protect chickens from various emerging clades of H5Nx avian influenza viruses.
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Krammer F, Smith GJD, Fouchier RAM, Peiris M, Kedzierska K, Doherty PC, Palese P, Shaw ML, Treanor J, Webster RG et al (2018) Influenza. Nat Rev Dis Primers 4:3
Falchi A (2020) Influenza D virus: the most discreet (for the moment?) of the influenza viruses. J Clin Med 9:E2550
Webster RG, Bean WJ, Gorman OT, Chambers TM, Kawaoka Y (1992) Evolution and ecology of influenza A viruses. Microbiol Rev 56:152–179
Tong S, Zhu X, Li Y, Shi M, Zhang J, Bourgeois M, Yang H, Chen X, Recuenco S, Gomez J et al (2013) New world bats harbor diverse influenza A viruses. PLoS Pathog 9:e1003657
Verhagen JH, Poen M, Stallknecht DE, van der Vliet S, Lexmond P, Sreevatsan S, Poulson RL, Fouchier RAM, Lebarbenchon C (2020) Phylogeography and Antigenic diversity of low-pathogenic avian influenza H13 and H16 viruses. J Virol 94:e00537-e1520
Lindsay LL, Plancarte M, Brenn-White M, Boyce WM (2014) Complete genome sequences of the first reported California h16 influenza A viruses. Genome Announc 2:e00329-e1314
Sauter NK, Bednarski MD, Wurzburg BA, Hanson JE, Whitesides GM, Skehel JJ, Wiley DC (1989) Hemagglutinins from two influenza virus variants bind to sialic acid derivatives with millimolar dissociation constants: a 500-MHz proton nuclear magnetic resonance study. Biochemistry 28:8388–8396
Costa T, Chaves AJ, Valle R, Darji A, Riel DV, Kuiken T, Majó N, Ramis A (2012) Distribution patterns of influenza virus receptors and viral attachment patterns in the respiratory and intestinal tracts of seven avian species. Vet Res 43:28
Qu N, Zhao B, Chen Z, He Z, Li W, Liu Z, Wang X, Huang J, Zhang Y, He W et al (2019) Genetic characteristics, pathogenicity and transmission of H5N6 highly pathogenic avian influenza viruses in Southern China. Transbound Emerg Dis 66:2411–2425
Wan XF (2012) Lessons from emergence of A/goose/Guangdong/1996-like H5N1 highly pathogenic avian influenza viruses and recent influenza surveillance efforts in southern China. Zoonoses Public Health 59:32–42
Suarez DL, Pantin-Jackwood MJ (2017) Recombinant viral-vectored vaccines for the control of avian influenza in poultry. Vet Microbiol 206:144–151
Liang WS, He YC, Wu HD, Li YT, Shih TH, Kao GS, Guo HY, Chao DY (2020) Ecological factors associated with persistent circulation of multiple highly pathogenic avian influenza viruses among poultry farms in Taiwan during 2015–2017. PLoS ONE 15:e0236581
Spackman E, Prosser DJ, Pantin-Jackwood M, Stephens CB, Berlin AM (2019) Clade 2.3.4.4 H5 north american highly pathogenic avian influenza viruses infect, but do not cause clinical signs in, American Black Ducks (Anas rubripes). Avian Dis 63:366–370
Marchenko V, Goncharova N, Susloparov I, Kolosova N, Gudymo A, Svyatchenko S, Danilenko A, Durymanov A, Gavrilova E, Maksyutov R et al (2018) Isolation and characterization of H5Nx highly pathogenic avian influenza viruses of clade 2.3.4.4 in Russia. Virology 525:216–223
Zhao K, Gu M, Zhong L, Duan Z, Zhang Y, Zhu Y, Zhao G, Zhao M, Chen Z, Hu S et al (2013) Characterization of three H5N5 and one H5N8 highly pathogenic avian influenza viruses in China. Vet Microbiol 163:351–357
Ku KB, Park EH, Yum J, Kim JA, Oh SK, Seo SH (2014) Highly pathogenic avian influenza A(H5N8) virus from waterfowl, South Korea, 2014. Emerg Infect Dis 20:1587–1588
Jiang W, Li Z, Liu S, Li J, Wang Y, Li J, Peng C, Song Q, Zhang L, Zhang F et al (2020) Genetic characterization of a highly pathogenic H5N6 avian influenza virus isolated from greylag goose. J Infect S0163–4453:30550–30558
Gomaa MR, El Rifay AS, Abu Zeid D, Elabd MA, Elabd E, Kandeil A, Shama NMA, Kamel MN, Marouf MA, Barakat A et al (2020) Incidence and seroprevalence of avian influenza in a cohort of backyard poultry growers, Egypt, August 2015–March 2019. Emerg Infect Dis 26:2129–2136
Ge Z, Gu M, Cai T, Liu K, Gao R, Liu D, Sun W, Li X, Shi L, Liu J et al (2021) Phylogenetic tracing and biological characterization of a novel clade 2.3.2.1 reassortant of H5N6 subtype avian influenza virus in China. Transbound Emerg Dis 68(2):730–741
Chen H (2009) Avian influenza vaccination: the experience in China. Rev Sci Tech 28:267–274
Swayne DE (2012) Impact of vaccines and vaccination on global control of avian influenza. Avian Dis 56:818–828
Bublot M, Pritchard N, Swayne DE, Selleck P, Karaca K, Suarez DL, Audonnet JC, Mickle TR (2006) Development and use of fowlpox vectored vaccines for avian influenza. Ann N Y Acad Sci 1081:193–201
Kapczynski DR, Esaki M, Dorsey KM, Jiang H, Jackwood M, Moraes M, Gardin Y (2015) Vaccine protection of chickens against antigenically diverse H5 highly pathogenic avian influenza isolates with a live HVT vector vaccine expressing the influenza hemagglutinin gene derived from a clade 2.2 avian influenza virus. Vaccine 33:1197–1205
Kim SH, Samal SK (2019) Innovation in Newcastle disease virus vectored avian influenza vaccines. Viruses 11:300
Richard-Mazet A, Goutebroze S, Le Gros FX, Swayne DE, Bublot M (2014) Immunogenicity and efficacy of fowlpox-vectored and inactivated avian influenza vaccines alone or in a prime-boost schedule in chickens with maternal antibodies. Vet Res 45:107
Schultz-Cherry S, Dybing JK, Davis NL, Williamson C, Suarez DL, Johnston R, Perdue ML (2000) Influenza virus (A/HK/156/97) hemagglutinin expressed by an alphavirus replicon system protects chickens against lethal infection with Hong Kong-origin H5N1 viruses. Virology 278:55–59
Reed LJ, Muench H (1938) A simple method of estimating fifty percent endpoints. Am J Hyg 27:493–497
Zhang X, Liu M, Liu C, Du J, Shi W, Sun E, Li H, Li J, Zhang Y (2011) Vaccination with different M2e epitope densities confers partial protection against H5N1 influenza A virus challenge in chickens. Intervirology 54:290–299
Ghorbani A, Ngunjiri JM, Xia M, Elaish M, Jang H, Mahesh KC, Abundo MC, Jiang X, Lee CW (2019) Heterosubtypic protection against avian influenza virus by live attenuated and chimeric norovirus P-particle-M2e vaccines in chickens. Vaccine 37:1356–1364
Elaish M, Kang KI, Xia M, Ali A, Shany SAS, Wang L, Jiang X, Lee CW (2015) Immunogenicity and protective efficacy of the norovirus P particle-M2e chimeric vaccine in chickens. Vaccine 33:4901–4909
Miller DS, Finnie J, Bowden TR, Scholz AC, Oh S, Kok T, Burrell CJ, Trinidad L, Boyle DB, Li P (2011) Preclinical efficacy studies of influenza A haemagglutinin precursor cleavage loop peptides as a potential vaccine. J Gen Virol 92:1152–1161
Tsai HJ, Chi LA, Yu AL (2012) Monoclonal antibodies targeting the synthetic peptide corresponding to the polybasic cleavage site on H5N1 influenza hemagglutinin. J Biomed Sci 19:37
Acknowledgements
This work was supported by a National Research Foundation of Korea (NRF) Grant funded by the Korean government (MSIT) (2019R1A2C200216812).
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The Chungnam National University (CNU) Internal Animal Use Committee approved the protocol (CNU-01191) for the pathogenicity study in chickens and the collection of clinical samples. All experiments and methods were performed with approval and in accordance with the respective relevant legal guidelines and regulations of Chungnam National University, Republic of Korea.
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Jang, Y., Seo, S.H. H5 cleavage-site peptide vaccine protects chickens from lethal infection by highly pathogenic H5 avian influenza viruses. Arch Virol 167, 67–75 (2022). https://doi.org/10.1007/s00705-021-05284-8
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DOI: https://doi.org/10.1007/s00705-021-05284-8