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Applied Microbiology and Biotechnology

, Volume 103, Issue 8, pp 3367–3379 | Cite as

Immunogenicity and protective efficacy of recombinant proteins consisting of multiple epitopes of foot-and-mouth disease virus fused with flagellin

  • Baofeng Cui
  • Xinsheng Liu
  • Peng Zhou
  • Yuzhen Fang
  • Donghong Zhao
  • Yongguang ZhangEmail author
  • Yonglu WangEmail author
Biotechnological products and process engineering
  • 138 Downloads

Abstract

Many recent studies have shown that flagellin fused to heterologous antigens can induce significantly enhanced humoral and cellular immune responses through its adjuvant activity. Therefore, in this study, two key B cell epitopes and a truncated VP1 (ΔVP1) protein from foot-and-mouth disease virus (FMDV) were expressed as flagellin fusion proteins in different patterns. Specifically, ΔVP1 and two duplicates of two key B cell epitopes (2×B1B2) were fused separately to the C-terminus of flagellin with a universal exogenous T cell epitope to construct FT (Flagellin-Truncated VP1) and FME (Flagellin-Multiple Epitopes). In addition, the D3 domain of flagellin was replaced by ΔVP1 in FME, yielding FTME (Flagellin-Truncated VP1-Multiple Epitopes). The immunogenicity and protective efficacy of the three fusion proteins as novel FMDV vaccine candidates were evaluated. The results showed that FT, FME, and FTME elicited significant FMDV-specific IgG responses at 10 μg/dose compared with the mock group (P < 0.05), with FTME producing the highest response. No significant differences in the antibody response to FTME were observed between different immunization routes or among adjuvants (ISA-206, poly(I·C), MPLA, and CpG-ODN) in mice. In addition, at 30 μg/dose, all three fusion proteins significantly induced neutralizing antibody production and upregulated the levels of some cytokines, including TNF-α, IFN-γ, and IL-12, in guinea pigs. Importantly, all three fusion proteins provided effective protective immunity against FMDV challenge in guinea pigs, though different protection rates were found. The results presented in this study indicate that the FTME fusion protein is a promising novel vaccine candidate for the future prevention and control of foot-and-mouth disease.

Keywords

FMDV Epitope Flagellin Recombinant fusion Vaccine 

Notes

Acknowledgements

This study was supported by the National Key Research and Development Program (2016YFD0501505), the Agriculture Research System of China (CARS-35), the Central Public Interest Scientific Institution Basal Research Fund (Y2016CG23), and the National Natural Science Foundation of China (31602095).

Compliance with ethical standards

Conflict of interests

The authors declare that they have no conflict of interest.

Ethics statement

All guinea pig experiments were performed in a biosafety level 3 laboratory at the Lanzhou Veterinary Research Institute (LVRI) at the Chinese Academy of Agricultural Sciences (CAAS). This study was approved by the Institutional Animal Use and Care Committee of the CAAS. All guinea pigs used in the present study were humanely bred during the experiments and were euthanized at the end of the study.

References

  1. Asadi KM, Oloomi M, Mahdavi M, Habibi M, Bouzari S (2013) Vaccination with recombinant FimH fused with flagellin enhances cellular and humoral immunity against urinary tract infection in mice. Vaccine 31(8):1210–1216.  https://doi.org/10.1016/j.vaccine.2012.12.059 CrossRefGoogle Scholar
  2. Aziz-Boaron O, Leibovitz K, Gelman B, Kedmi M, Klement E (2013) Safety, immunogenicity and duration of immunity elicited by an inactivated bovine ephemeral fever vaccine. PLoS One 8(12):e82217.  https://doi.org/10.1371/journal.pone.0082217 CrossRefGoogle Scholar
  3. Bargieri DY, Rosa DS, Braga CJ, Carvalho BO, Costa FT, Espindola NM, Vaz AJ, Soares IS, Ferreira LC, Rodrigues MM (2008) New malaria vaccine candidates based on the Plasmodium vivax merozoite surface protein-1 and the TLR-5 agonist Salmonella Typhimurium FliC flagellin. Vaccine 26(48):6132–6142.  https://doi.org/10.1016/j.vaccine.2008.08.070 CrossRefGoogle Scholar
  4. Barnett PV, Carabin H (2002) A review of emergency foot-and-mouth disease (FMD) vaccines. Vaccine 20(11–12):1505–1514CrossRefGoogle Scholar
  5. Bittle JL, Houghten RA, Alexander H, Shinnick TM, Sutcliffe JG, Lerner RA, Rowlands DJ, Brown F (1982) Protection against foot-and-mouth disease by immunization with a chemically synthesized peptide predicted from the viral nucleotide sequence. Nature 298(5869):30–33CrossRefGoogle Scholar
  6. Brown F (1988) Use of peptides for immunization against foot-and-mouth disease. Vaccine 6(2):180–182CrossRefGoogle Scholar
  7. Burman A, Clark S, Abrescia NG, Fry EE, Stuart DI, Jackson T (2006) Specificity of the VP1 GH loop of foot-and-mouth disease virus for alphav integrins. J Virol 80(19):9798–9810.  https://doi.org/10.1128/JVI.00577-06 CrossRefGoogle Scholar
  8. Capozzo AV, Wilda M, Bucafusco D, de Los ALM, Franco-Mahecha OL, Mansilla FC, Perez-Filgueira DM, Grigera PR (2011) Vesicular stomatitis virus glycoprotein G carrying a tandem dimer of foot and mouth disease virus antigenic site a can be used as DNA and peptide vaccine for cattle. Antivir Res 92(2):219–227.  https://doi.org/10.1016/j.antiviral.2011.08.006 CrossRefGoogle Scholar
  9. Chen YS, Hung YC, Lin WH, Huang GS (2010) Assessment of gold nanoparticles as a size-dependent vaccine carrier for enhancing the antibody response against synthetic foot-and-mouth disease virus peptide. Nanotechnology 21(19):195101.  https://doi.org/10.1088/0957-4484/21/19/195101 CrossRefGoogle Scholar
  10. Cui B, Liu X, Fang Y, Zhou P, Zhang Y, Wang Y (2018) Flagellin as a vaccine adjuvant. Expert Rev Vaccines 17(4):335–349.  https://doi.org/10.1080/14760584.2018.1457443 CrossRefGoogle Scholar
  11. Dicara D, Burman A, Clark S, Berryman S, Howard MJ, Hart IR, Marshall JF, Jackson T (2008) Foot-and-mouth disease virus forms a highly stable, EDTA-resistant complex with its principal receptor, integrin alphavbeta6: implications for infectiousness. J Virol 82(3):1537–1546.  https://doi.org/10.1128/JVI.01480-07 CrossRefGoogle Scholar
  12. Doel TR, Gale C, Do AC, Mulcahy G, Dimarchi R (1990) Heterotypic protection induced by synthetic peptides corresponding to three serotypes of foot-and-mouth disease virus. J Virol 64(5):2260–2264Google Scholar
  13. Dory D, Remond M, Beven V, Cariolet R, Backovic M, Zientara S, Jestin A (2009) Pseudorabies virus glycoprotein B can be used to carry foot and mouth disease antigens in DNA vaccination of pigs. Antivir Res 81(3):217–225.  https://doi.org/10.1016/j.antiviral.2008.11.005 CrossRefGoogle Scholar
  14. Du Y, Li Y, He H, Qi J, Jiang W, Wang X, Tang B, Cao J, Wang X, Jiang P (2008) Enhanced immunogenicity of multiple-epitopes of foot-and-mouth disease virus fused with porcine interferon alpha in mice and protective efficacy in guinea pigs and swine. J Virol Methods 149(1):144–152.  https://doi.org/10.1016/j.jviromet.2007.12.018 CrossRefGoogle Scholar
  15. Golde WT, Pacheco JM, Duque H, Doel T, Penfold B, Ferman GS, Gregg DR, Rodriguez LL (2005) Vaccination against foot-and-mouth disease virus confers complete clinical protection in 7 days and partial protection in 4 days: use in emergency outbreak response. Vaccine 23(50):5775–5782.  https://doi.org/10.1016/j.vaccine.2005.07.043 CrossRefGoogle Scholar
  16. Grubman MJ, Baxt B (2004) Foot-and-mouth disease. Clin Microbiol Rev 17(2):465–493CrossRefGoogle Scholar
  17. Honko AN, Sriranganathan N, Lees CJ, Mizel SB (2006) Flagellin is an effective adjuvant for immunization against lethal respiratory challenge with Yersinia pestis. Infect Immun 74(2):1113–1120.  https://doi.org/10.1128/IAI.74.2.1113-1120.2006 CrossRefGoogle Scholar
  18. Huleatt JW, Jacobs AR, Tang J, Desai P, Kopp EB, Huang Y, Song L, Nakaar V, Powell TJ (2007) Vaccination with recombinant fusion proteins incorporating Toll-like receptor ligands induces rapid cellular and humoral immunity. Vaccine 25(4):763–775.  https://doi.org/10.1016/j.vaccine.2006.08.013 CrossRefGoogle Scholar
  19. Huleatt JW, Nakaar V, Desai P, Huang Y, Hewitt D, Jacobs A, Tang J, McDonald W, Song L, Evans RK, Umlauf S, Tussey L, Powell TJ (2008) Potent immunogenicity and efficacy of a universal influenza vaccine candidate comprising a recombinant fusion protein linking influenza M2e to the TLR5 ligand flagellin. Vaccine 26(2):201–214.  https://doi.org/10.1016/j.vaccine.2007.10.062 CrossRefGoogle Scholar
  20. Khalifa ME, El-Deeb AH, Zeidan SM, Hussein HA, Abu-El-Naga HI (2017) Enhanced protection against FMDV in cattle after prime-boost vaccination based on mucosal and inactivated FMD vaccine. Vet Microbiol 210:1–7.  https://doi.org/10.1016/j.vetmic.2017.08.014 CrossRefGoogle Scholar
  21. Lawrence P, Pacheco JM, Uddowla S, Hollister J, Kotecha A, Fry E, Rieder E (2013) Foot-and-mouth disease virus (FMDV) with a stable FLAG epitope in the VP1 G-H loop as a new tool for studying FMDV pathogenesis. Virology 436(1):150–161.  https://doi.org/10.1016/j.virol.2012.11.001 CrossRefGoogle Scholar
  22. Li G, Chen W, Yan W, Zhao K, Liu M, Zhang J, Fei L, Xu Q, Sheng Z, Lu Y, Zheng Z (2004) Comparison of immune responses against foot-and-mouth disease virus induced by fusion proteins using the swine IgG heavy chain constant region or beta-galactosidase as a carrier of immunogenic epitopes. Virology 328(2):274–281.  https://doi.org/10.1016/j.virol.2004.07.025 CrossRefGoogle Scholar
  23. Li K, Bao H, Wei G, Li D, Chen Y, Fu Y, Cao Y, Li P, Sun P, Bai X, Ma X, Zhang J, Lu Z, Liu Z (2017) Molecular vaccine prepared by fusion of XCL1 to the multi-epitope protein of foot-and-mouth disease virus enhances the specific humoural immune response in cattle. Appl Microbiol Biotechnol 101(21):7889–7900.  https://doi.org/10.1007/s00253-017-8523-y CrossRefGoogle Scholar
  24. Liu X, Fang Y, Zhou P, Lu Y, Zhang Q, Xiao S, Dong Z, Pan L, Lv J, Zhang Z, Zhang Y, Wang Y (2017) Chimeric virus-like particles elicit protective immunity against serotype O foot-and-mouth disease virus in guinea pigs. Appl Microbiol Biotechnol 101(12):4905–4914.  https://doi.org/10.1007/s00253-017-8246-0 CrossRefGoogle Scholar
  25. Manicassamy S, Pulendran B (2009) Modulation of adaptive immunity with Toll-like receptors. Semin Immunol 21(4):185–193.  https://doi.org/10.1016/j.smim.2009.05.005 CrossRefGoogle Scholar
  26. McDonald WF, Huleatt JW, Foellmer HG, Hewitt D, Tang J, Desai P, Price A, Jacobs A, Takahashi VN, Huang Y, Nakaar V, Alexopoulou L, Fikrig E, Powell TJ (2007) A West Nile virus recombinant protein vaccine that coactivates innate and adaptive immunity. J Infect Dis 195(11):1607–1617.  https://doi.org/10.1086/517613 CrossRefGoogle Scholar
  27. Meloen RH, Barteling SJ (1986) An epitope located at the C terminus of isolated VP1 of foot-and-mouth disease virus type O induces neutralizing activity but poor protection. J Gen Virol 67(Pt 2):289–294.  https://doi.org/10.1099/0022-1317-67-2-289. CrossRefGoogle Scholar
  28. Mizel SB, Honko AN, Moors MA, Smith PS, West AP (2003) Induction of macrophage nitric oxide production by Gram-negative flagellin involves signaling via heteromeric Toll-like receptor 5/Toll-like receptor 4 complexes. J Immunol 170(12):6217–6223CrossRefGoogle Scholar
  29. Morgan DO, Moore DM (1990) Protection of cattle and swine against foot-and-mouth disease, using biosynthetic peptide vaccines. Am J Vet Res 51(1):40–45Google Scholar
  30. Mount A, Koernig S, Silva A, Drane D, Maraskovsky E, Morelli AB (2013) Combination of adjuvants: the future of vaccine design. Expert Rev Vaccines 12(7):733–746.  https://doi.org/10.1586/14760584.2013.811185 CrossRefGoogle Scholar
  31. Pandey JP (2011) Comment on “Flagellin as an adjuvant: cellular mechanisms and potential”. J Immunol 186(3):1299; author reply 1299.  https://doi.org/10.4049/jimmunol.1090134 CrossRefGoogle Scholar
  32. Robinson L, Knight-Jones TJ, Charleston B, Rodriguez LL, Gay CG, Sumption KJ, Vosloo W (2016) Global foot-and-mouth disease research update and gap analysis: 3—vaccines. Transbound Emerg Dis 63(Suppl 1):30–41.  https://doi.org/10.1111/tbed.12521. CrossRefGoogle Scholar
  33. Rodriguez LL, Grubman MJ (2009) Foot and mouth disease virus vaccines. Vaccine 27(Suppl 4):D90–D94.  https://doi.org/10.1016/j.vaccine.2009.08.039. CrossRefGoogle Scholar
  34. Shao JJ, Wang JF, Chang HY, Liu JX (2011) Immune potential of a novel multiple-epitope vaccine to FMDV type Asia 1 in guinea pigs and sheep. Virol Sin 26(3):190–197.  https://doi.org/10.1007/s12250-011-3174-0 CrossRefGoogle Scholar
  35. Song L, Nakaar V, Kavita U, Price A, Huleatt J, Tang J, Jacobs A, Liu G, Huang Y, Desai P, Maksymiuk G, Takahashi V, Umlauf S, Reiserova L, Bell R, Li H, Zhang Y, McDonald WF, Powell TJ, Tussey L (2008) Efficacious recombinant influenza vaccines produced by high yield bacterial expression: a solution to global pandemic and seasonal needs. PLoS One 3(5):e2257.  https://doi.org/10.1371/journal.pone.0002257 CrossRefGoogle Scholar
  36. Steinhagen F, Kinjo T, Bode C, Klinman DM (2011) TLR-based immune adjuvants. Vaccine 29(17):3341–3355.  https://doi.org/10.1016/j.vaccine.2010.08.002 CrossRefGoogle Scholar
  37. Taylor DN, Treanor JJ, Sheldon EA, Johnson C, Umlauf S, Song L, Kavita U, Liu G, Tussey L, Ozer K, Hofstaetter T, Shaw A (2012) Development of VAX128, a recombinant hemagglutinin (HA) influenza-flagellin fusion vaccine with improved safety and immune response. Vaccine 30(39):5761–5769.  https://doi.org/10.1016/j.vaccine.2012.06.086 CrossRefGoogle Scholar
  38. Wang G, Pan L, Zhang Y, Wang Y, Zhang Z, Lu J, Zhou P, Fang Y, Jiang S (2011) Intranasal delivery of cationic PLGA nano/microparticles-loaded FMDV DNA vaccine encoding IL-6 elicited protective immunity against FMDV challenge. PLoS One 6(11):e27605.  https://doi.org/10.1371/journal.pone.0027605 CrossRefGoogle Scholar
  39. Wen X, Wen K, Cao D, Li G, Jones RW, Li J, Szu S, Hoshino Y, Yuan L (2014) Inclusion of a universal tetanus toxoid CD4(+) T cell epitope P2 significantly enhanced the immunogenicity of recombinant rotavirus DeltaVP8* subunit parenteral vaccines. Vaccine 32(35):4420–4427.  https://doi.org/10.1016/j.vaccine.2014.06.060 CrossRefGoogle Scholar
  40. Yoon SI, Kurnasov O, Natarajan V, Hong M, Gudkov AV, Osterman AL, Wilson IA (2012) Structural basis of TLR5-flagellin recognition and signaling. Science 335(6070):859–864.  https://doi.org/10.1126/science.1215584 CrossRefGoogle Scholar
  41. Zamorano P, Wigdorovitz A, Perez-Filgueira M, Carrillo C, Escribano JM, Sadir AM, Borca MV (1995) A 10-amino-acid linear sequence of VP1 of foot and mouth disease virus containing B- and T-cell epitopes induces protection in mice. Virology 212(2):614–621.  https://doi.org/10.1006/viro.1995.1519 CrossRefGoogle Scholar
  42. Zhang YL, Guo YJ, Wang KY, Lu K, Li K, Zhu Y, Sun SH (2007) Enhanced immunogenicity of modified hepatitis B virus core particle fused with multiepitopes of foot-and-mouth disease virus. Scand J Immunol 65(4):320–328.  https://doi.org/10.1111/j.1365-3083.2007.01900.x CrossRefGoogle Scholar
  43. Zhang C, Zhu S, Wei L, Yan X, Wang J, Quan R, She R, Hu F, Liu J (2015) Recombinant flagellin-porcine circovirus type 2 cap fusion protein promotes protective immune responses in mice. PLoS One 10(6):e0129617.  https://doi.org/10.1371/journal.pone.0129617 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Baofeng Cui
    • 1
    • 2
    • 3
  • Xinsheng Liu
    • 1
    • 2
  • Peng Zhou
    • 1
    • 2
  • Yuzhen Fang
    • 1
    • 2
  • Donghong Zhao
    • 1
    • 2
  • Yongguang Zhang
    • 1
    • 2
    Email author
  • Yonglu Wang
    • 1
    • 2
    Email author
  1. 1.State Key Laboratory of Veterinary Etiological Biology, OIE/National Foot and Mouth Disease Reference Laboratory, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research InstituteChinese Academy of Agricultural SciencesLanzhouChina
  2. 2.Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and ZoonosesYangzhouChina
  3. 3.Lanzhou Institute of Biological Products Co., Ltd. (LIBP), a subsidiary company of China National Biotec Group Company Limited (CNBG)LanzhouChina

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