Top

Published in:

Open Access 01-12-2014 | Research

Heterosubtypic protective immunity against widely divergent influenza subtypes induced by fusion protein 4sM2 in BALB/c mice

Authors: Mohammed YE Chowdhury, Soo-Kyung Seo, Ho-Jin Moon, Melbourne R Talactac, Jae-Hoon Kim, Min-Eun Park, Hwa-Young Son, Jong-Soo Lee, Chul-Joong Kim

Published in: Virology Journal | Issue 1/2014

Abstract

Background

Regular reformulation of currently available vaccines is necessary due to the unpredictable variability of influenza viruses. Therefore, vaccine based on a highly conserved antigen with capability of induction of effective immune responses could be a potential solution. Influenza matrix protein-2 (M2) is highly conserved across influenza subtypes and a promising candidate for a broadly protective influenza vaccine. For the enhancement of broad protection, four tandem copies of consensus M2 gene containing extracellular (ED) and cytoplasmic (CD) without the trans-membrane domain (TM) reconstituted from H1N1, H5N1 and H9N2 influenza viruses were linked and named as 4sM2. The construct was effectively expressed in Escherichia coli, purified and proteins were used to immunize BALB/c mice. Humoral and cell-mediated immune responses were investigated following administration.

Results

Mice were intramuscularly immunized with 4sM2 protein 2 times at 2 weeks interval. Two weeks after the last immunization, first humoral and cell mediated immune response specific to sM2 protein were evaluated and the mice were challenged with a lethal dose (10MLD₅₀) of divergent subtypes A/EM/Korea/W149/06(H5N1), A/PR/8/34(H1N1), A/Aquatic bird/Korea/W81/2005(H5N2), A/Aquatic bird/Korea/W44/2005(H7N3), and A/Chicken/Korea/116/2004(H9N2) viruses. The efficacy of 4sM2 was evaluated by determining survival rates, body weights and residual lung viral titers. Our studies demonstrate that the survival of mice immunized with 4sM2 was significantly higher (80–100% survival) than that of unimmunized mice (0% survival). We also examined the long lasting protection against heterosubtype H5N2 virus and found that mice vaccinated with 4sM2 displayed 80% of protection even after 6 months of final vaccination.

Conclusion

Taken together, these results suggest that prokaryotic expressed multimeric sM2 protein achieved cross protection against lethal infection of divergent influenza subtypes which are lasting for the long time.

Available only for authorised users

Zimmer SM, Burke DS: Historical perspective - emergence of influenza A (H1N1) viruses. N Engl J Med 2009, 361: 279-285. 10.1056/NEJMra0904322PubMedCrossRef

Stephenson I, Nicholson KG: Influenza: vaccination and treatment. Eur Respir J 2001, 17: 1282-1293. 10.1183/09031936.01.00084301PubMedCrossRef

Couch RB: Seasonal inactivated influenza virus vaccines. Vaccine 2008, 26S: 5-9.CrossRef

Al-Marou A, Scheifele DW, Soong T, Bjornson G: Comparison of adverse reactions to whole-virion and split-virion influenza vaccines in hospital personnel. Can Med Assoc J 1991, 145: 213-218.

Song JM, Rooijen NV, Bozja J, Compans RW, Kang SM: Vaccination inducing broad and improved cross protection against multiple subtypes of influenza A virus. PNAS 2011, 108: 757-761. 10.1073/pnas.1012199108PubMedPubMedCentralCrossRef

Jennings R, Smith TL, Spencer RC, Mellersh AM, Edey D, Fenton P, Potter CW: Inactivated influenza virus vaccines in man: a comparative study of subunit and split vaccines using two methods for assessment of antibody responses. Vaccine 1984, 2: 75-80. 10.1016/S0264-410X(98)90036-6PubMedCrossRef

Beyer WEP, Palache AM, Jong JCD, Osterhaus ADME: Cold-adapted live influenza vaccine versus inactivated vaccine: systemic vaccine reactions, local and systemic antibody response, and vaccine efficacy. A meta-analysis. Vaccine 2002, 20: 1340-1353. 10.1016/S0264-410X(01)00471-6PubMedCrossRef

Gerdil C: The annual production cycle for influenza vaccine. Vaccine 2003, 21: 1776-1779. 10.1016/S0264-410X(03)00071-9PubMedCrossRef

Smith DJ, Lapedes AS, Jong JCD, Bestebroer TM, Rimmelzwaan GF, Osterhaus ADME, Fouchier RAM: Mapping the antigenic and genetic evolution of influenza virus. Science 2004, 305: 371-376. 10.1126/science.1097211PubMedCrossRef

10.

Kang SM, Song JM, Compans RW: Novel vaccines against influenza viruses. Virus Res 2011, 162: 31-38. 10.1016/j.virusres.2011.09.037PubMedPubMedCentralCrossRef

11.

Zhao G, Lin Y, Du I, Guan J, Sun S, Sui H, Kou Z, Chan CC, Guo Y, Jiang S, Zheng BJ, Zhou Y: An M2e-based multiple antigenic peptide vaccine protects mice from lethal challenge with divergent H5N1 influenza viruses. Virol J 2010, 7: 2-8. 10.1186/1743-422X-7-2CrossRef

12.

Lalor PA, Webby RJ, Morrow J, Rusalov D, Kaslow DC, Rolland A, Smith LR: Plasmid DNA based vaccines protect mice and ferrets against lethal challenge with A/Vietnam/1203/04 (H5N1) influenza virus. J Infect Dis 2008, 197: 1643-1652. 10.1086/588431PubMedCrossRef

13.

Huleatt JW, Nakaara V, Desaia P, Huanga Y, Hewitt D, Jacobsa A, Tanga J, McDonalda W, Songb L, Evans RK, Umlaufb S, Tusseyb L, Powella TJ: Potent immunogenicity and efficacy of a universal influenza vaccine candidate comprising are combinant fusion protein linking influenza M2e to the TLR5 ligand flagellin. Vaccine 2008, 26: 201-214. 10.1016/j.vaccine.2007.10.062PubMedCrossRef

14.

Wu F, Huang JH, Yuan XY, Huang WS, Chen YH: Characterization of immunity induced by M2e of influenza virus. Vaccine 2007, 25: 8868-8873. 10.1016/j.vaccine.2007.09.056PubMedCrossRef

15.

Ernst WA, Kim HJ, Tumpey TM, Jansen AD, Tai W, Cramer DV, Adler-Moore JP, Fujii G: Protection against H1, H5, H6 and H9 influenza A infection with Liposomal matrix 2 epitope vaccines. Vaccine 2006, 24: 5158-5168. 10.1016/j.vaccine.2006.04.008PubMedCrossRef

16.

Tompkins SM, Zhao ZS, Lo CY, Misplon JA, Liu T, Ye Z, Hogan RJ, Wu Z, Benton KA, Tumpey TM, Epstein SL: Matrix protein 2 vaccination and protection against influenza viruses, including subtype H5N1. Emerg Infect Dis 2007, 13: 426-435. 10.3201/eid1303.061125PubMedPubMedCentralCrossRef

17.

Jimenez GS, Planchon R, Wei Q, Rusalov D, Geall A, Enas J, Lalor P, Leamy V, Vahle R, Luke CJ, Rolland A, Kaslow DC, Smith LR: Vaxfectin formulated influenza DNA vaccines encoding NP and M2 viral proteins protect mice against lethal viral challenge. Hum Vaccin 2007, 3: 157-164. 10.4161/hv.3.5.4175PubMedCrossRef

18.

Song JM, Wang BZ, Park KM, Rooijen NV, Quan FS, Kim MC, Jin HT, Pekosz A, Compans RW, Kang SM: Influenza virus-like particles containing M2 induce broadly cross protective immunity. PLoS One 2011, 6: e14538. 10.1371/journal.pone.0014538PubMedPubMedCentralCrossRef

19.

Schnell JR, Chou JJ: Structure and mechanism of the M2 proton channel of influenza A virus. Nature 2008, 451: 591-595. 10.1038/nature06531PubMedPubMedCentralCrossRef

20.

Du L, Zhou Y, Jiang S: Research and development of universal influenza vaccines. Microbes Infect 2010, 12: 280-286. 10.1016/j.micinf.2010.01.001PubMedCrossRef

21.

Liu W, Zou P, Chen YH: Monoclonal antibodies recognizing EVETPIRN epitope of influenza A virus M2 protein could protect mice from lethal influenza A virus challenge. Immunol Lett 2004, 93: 131-136. 10.1016/j.imlet.2004.03.003PubMedCrossRef

22.

Zou P, Liu W, Chen YH: The epitope recognized by a monoclonal antibody in influenza A virus M2 protein is immunogenic and confers immune protection. Int Immunopharmacol 2005, 5: 631-635. 10.1016/j.intimp.2004.12.005PubMedCrossRef

23.

Moon HJ, Lee JS, Talactac MR, Chowdhury MYE, Kim JH, Park ME, Choi YK, Sung MH, Kim CJ: Mucosal immunization with recombinant influenza hemagglutinin protein and poly gamma-glutamate/chitosan nanoparticles induces protection against highly pathogenic influenza A virus. Vet Microbiol 2012, 160: 277-289. 10.1016/j.vetmic.2012.05.035PubMedCrossRef

24.

Zhao G, Du L, Xiao W, Sun S, Lin Y, Chen M, Kou Z, He Y, Lustigman S, Jiang S, Zheng BJ, Zhou Y: Induction of the protection against divergent H5N1 influenza viruses using a recombinant fusion protein linking influenza M2e to onchocerca volvulus activation associated protein-1 (ASP-1) adjuvant. Vaccine 2010, 28: 7233-7240. 10.1016/j.vaccine.2010.08.049PubMedCrossRef

25.

Slepushkin VA, Katz JM, Black RA, Gamble WC, Rota PA, Cox NJ: Protection of mice against influenza A virus challenge by vaccination with baculovirus-expressed M2 protein. Vaccine 1995, 13: 1399-1402. 10.1016/0264-410X(95)92777-YPubMedCrossRef

26.

Ebrahimi SM, Dabaghian M, Tebianian M, Jazi MHZ: In contrast to conventional inactivated influenza vaccines, 4×M2e.HSP70c fusion protein fully protected mice against lethal dose of H1, H3 and H9 influenza A isolates circulating in Iran. Virology 2012, 430: 63-72. 10.1016/j.virol.2012.04.015PubMedCrossRef

27.

Alvarez P, Zylberman V, Ghersi G, Boado L, Palacios C, Goldbaum F, Mattion N: Tandem repeats of the extracellular domain of Matrix 2 influenza protein exposed in Brucella lumazine synthase decameric carrier molecule induce protection in mice. Vaccine 2013, 31: 806-812. 10.1016/j.vaccine.2012.11.072PubMedCrossRef

28.

Kim MC, Song JM, Eunju O, Kwon YM, Lee YJ, Compans RW, Kang SM: Virus-like particles containing multiple M2 extracellular domains confer improved cross-protection against various subtypes of influenza virus. Mol Ther 2013, 21: 485-492. 10.1038/mt.2012.246PubMedPubMedCentralCrossRef

29.

Jegerlehner A, Schmitz N, Storni T, Bachmann MF: Influenza A vaccine based on the extracellular domain of M2: weak protection mediated via antibody-dependent NK cell activity. J Immunol 2004, 172: 5598-5605.PubMedCrossRef

30.

Wu F, Yuan XY, Li J, Chen YH: The co-administration of CpG-ODN influenced protective activity of influenza M2e vaccine. Vaccine 2009, 27: 4320-4324. 10.1016/j.vaccine.2009.04.075PubMedCrossRef

31.

Sui J, Hwang WC, Perez S, Wei G, Aird D, Chen LM, Santelli E, Stec B, et al.: Structural and functional bases for broad-spectrum neutralization of avian and human influenza A viruses. Nat Struct Mol Biol 2009, 16: 265-273. 10.1038/nsmb.1566PubMedPubMedCentralCrossRef

32.

Price GE, Soboleski MR, Lo CY, Misplon JA, Pappas C: Vaccination focusing immunity on conserved antigens protects mice and ferrets against virulent H1N1 and H5N1 influenza A viruses. Vaccine 2009, 27: 6512-6521. 10.1016/j.vaccine.2009.08.053PubMedCrossRef

33.

Cox NJ, Subbarao K: Influenza. Lancet 1999, 354: 1277-1282. 10.1016/S0140-6736(99)01241-6PubMedCrossRef

34.

Ghedin E, Sengamalay NA, Shumway M, Zaborsky J, Feldblyum T, Subbu V, Spiro DJ, Sitz J, Koo H, Bolotov P, Dernovoy D, Tatusova T, Bao Y, George KS, Taylor J, Lipman DJ, Fraser CM, Taubenberger JK, Salzberg SL: Large-scale sequencing of human influenza reveals the dynamic nature of viral genome evolution. Nature 2005, 437: 1162-1166. 10.1038/nature04239PubMedCrossRef

35.

Oshitani H, Kamigaki T, Suzuki A: Major issues and challenges of influenza pandemic preparedness in developing countries. Emerg Infect Dis 2008, 14: 875-880. 10.3201/eid1406.070839PubMedPubMedCentralCrossRef

36.

Sui Z, Chen Q, Wu R, Zhang H, Zheng M, Wang H, Chen Z: Cross-protection against influenza virus infection by intranasal administration of M2-based vaccine with chitosan as an adjuvant. Arch Virol 2010, 155: 535-544. 10.1007/s00705-010-0621-4PubMedCrossRef

37.

Ekiert DC, Bhabha G, Elsliger MA, Friesen RHE, Jongeneelen M, Throsby M, Goudsmit J, Wilson IA: Antibody recognition of a highly conserved influenza virus epitop. Science 2009, 324: 246-251. 10.1126/science.1171491PubMedPubMedCentralCrossRef

38.

Lee JS, Chowdhury MYE, Moon HJ, Choi YK, Talactac MR, Kim JH, Park ME, Son HY, Shin KH, Kim CJ: The highly conserved HA2 protein of the influenza a virus induces a cross protective immune response. J Virol Methods 2013, 194: 280-288. 10.1016/j.jviromet.2013.08.022PubMedCrossRef

39.

Zhang T, Xu X, Shen L, Feng Y, Yang Z, Shen Y, Wang J, In W, Wang X: Modeling of protein refolding from inclusion bodies. Acta Biochim Biophys Sin 2009, 4: 1044-1052.CrossRef

40.

Shim BS, Choi YK CH, Yun Lee EG, Jeon YS, Park SM, Cheon IS, Joo DH, Cho CH, Song MS, et al.: Sublingual immunization with M2-based vaccine induces broad protective immunity against influenza. PLoS One 2011, 6: e27953. 10.1371/journal.pone.0027953PubMedPubMedCentralCrossRef

41.

Okuda K, Ihata A, Watabe S, Okada E, Yamakawa T, Hamajima K, Yang J, Ishii N, Nakazawa M, Okuda K, Ohnari K, Nakajima K, Xin KQ: Protective immunity against influenza A virus induced by immunization with DNA plasmid containing influenza M gene. Vaccine 2001, 19: 3681-3691. 10.1016/S0264-410X(01)00078-0PubMedCrossRef

42.

Oran AE, Robinson HL: DNA vaccine combining form of antigen and method of delivery to raise a spectrum of IFN-gamma and IL-4 producing CD4+ and CD8+ T cells. J Immunol 2003, 171: 1999-2005.PubMedCrossRef

43.

Price GE, Soboleski MR, Lo CY, Misplon JA, Quirion MR, Houser KV, Pearce MB, Pappas C, Tumpey TM, Epstein SL: Single-dose mucosal immunization with a candidate universal influenza vaccine provides rapid protection from virulent H5N1, H3N2 and H1N1 viruses. PLoS One 2010, 5: e13162. 10.1371/journal.pone.0013162PubMedPubMedCentralCrossRef

44.

Chen J, Zhang F, Fang F, Chang H, Chen Z: Vaccination with hemagglutinin or neuraminidase DNA protects BALB/c mice against influenza virus infection in presence of maternal antibody. BMC Infect Dis 2007, 7: 118. 10.1186/1471-2334-7-118PubMedPubMedCentralCrossRef

45.

Qiu M, Fang F, Chen Y, Wang H, Chen Q, Chang H, Wang F, et al.: Protection against avian influenza H9N2 virus challenge by immunization with hemagglutinin or neuraminidase-expressing DNA in BALB/c mice. Biochem Biophys Res Commun 2006, 343: 1124-1131. 10.1016/j.bbrc.2006.03.088PubMedCrossRef

46.

Itoh Y, Shinya K, Kiso M: In vitro and in vivo characterization of new swine-origin H1N1 influenza viruses. Nature 2009, 460: 1021-1027.PubMedPubMedCentral

Title: Heterosubtypic protective immunity against widely divergent influenza subtypes induced by fusion protein 4sM2 in BALB/c mice
Authors: Mohammed YE Chowdhury
Soo-Kyung Seo
Ho-Jin Moon
Melbourne R Talactac
Jae-Hoon Kim
Min-Eun Park
Hwa-Young Son
Jong-Soo Lee
Chul-Joong Kim
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Virology Journal / Issue 1/2014
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/1743-422X-11-21

ACC 2024 Congress Coverage

Heart Failure Video

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Heterosubtypic protective immunity against widely divergent influenza subtypes induced by fusion protein 4sM2 in BALB/c mice

Abstract

Background

Results

Conclusion

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2014

Comparison of multiple DNA vaccines for protection against cytomegalovirus infection in BALB/c mice

Small molecules that inhibit Vif-induced degradation of APOBEC3G

Pandemic clinical case definitions are non-specific: multiple respiratory viruses circulating in the early phases of the 2009 influenza pandemic in New South Wales, Australia

Spontaneous reactivation of hepatitis B virus replication in an HIV coinfected patient with isolated anti-Hepatitis B core antibodies

Genomic variation in macrophage-cultured European porcine reproductive and respiratory syndrome virus Olot/91 revealed using ultra-deep next generation sequencing

Inhibition of hepatitis C virus by an M1GS ribozyme derived from the catalytic RNA subunit of Escherichia coli RNase P

ACC 2024 Congress Coverage

Year in Review: Heart failure and cardiomyopathies

Semaglutide benefits people with type 2 diabetes and obesity-related heart failure

Incentivised to exercise

New intervention for STEMI-related cardiogenic shock

» View more highlights on the ACC 2024 congress hub page