Skip to main content

Advertisement

SpringerLink
Log in

TLR-5 agonist Salmonella abortus equi flagellin FliC enhances FliC-gD-based DNA vaccination against equine herpesvirus 1 infection

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Equine herpesvirus 1 (EHV-1) induces serious respiratory infections, viral abortion, neurological signs, and neonatal mortality in horses. Despite the use of vaccines, EHV-1 infection also causes a high annual economic burden to the equine industry. The poor immunogenicity of and protection conferred by EHV-1 vaccines are the major factors responsible for the spread of EHV-1 infection. The present study examined the immunogenicity of a novel DNA vaccine co-expressing FliC, a flagellin protein, in Salmonella abortus equi and the gD protein of EHV-1. Mice and horses were immunized intramuscularly with the vaccine, and mice were challenged with EHV-1. Immunofluorescence and western blotting revealed that FliC and gD can be efficiently expressed in cells. This novel vaccine significantly increased gD-specific antibody and interferon gamma (IFN-γ) levels in immunized mice and horses. Compared with controls, the viral load and morbidity were markedly reduced in FliC-gD-immunized mice after they were challenged with EHV-1. Furthermore, the immunogenicity of FliC-gD in a natural host was tested. Our results indicate that vaccinated mice and horses exhibit increased humoral and improved cellular immune responses.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Crabb BS, Studdert MJ (1995) Equine herpesviruses 4 (equine rhinopneumonitis virus) and 1 (equine abortion virus). Adv Virus Res 45:153–190

    Article  CAS  PubMed  Google Scholar 

  2. Reed SM, Toribo RE (2004) Equine herpesvirus 1 and 4. Vet Clin Am Equine Pract 20:631–642

    Article  Google Scholar 

  3. Allen GP, Bolin DC, Bryant U, Carter CN, Giles RC, Harrison LR, Hong CB, Jackson CB, Poonacha K, Wharton R, Williams NM (2008) Prevalence of latent, neuropathogenic equine herpesvirus-1 in the thoroughbred broodmare population of central kentucky. Equine Vet J 40:105–110

    Article  CAS  PubMed  Google Scholar 

  4. Davis-Poynter NJ, Farrell HE (1996) Masters of deception:a review of herpesvirus immune evasion strategies. Immunol Cell Biol 74:513–522

    Article  CAS  PubMed  Google Scholar 

  5. Gilkerson JR, Love DN, Whalley JM (1997) Serological evidence of equineherpesvirus 1 (EHV-1) infection in thoroughbred foals 30–120 days of age. Aust Vet J 15:128–134

    Google Scholar 

  6. Patel JR, Heldens J (2005) Equine herpesviruses 1 (EHV-1) and 4 (EHV-4)-epidemiology, disease and immunoprophylaxis: a brief review. Vet J 170:14–23

    Article  CAS  Google Scholar 

  7. Van MC, Willink DL, Smeenk LA, Brinkhof J, Terpstra C (2000) An equine herpesvirus 1 (EHV1) abortion storm at a riding school. Vet Q 22:83–87

    Article  Google Scholar 

  8. Csellner H, Walker C, Wellington JE, Mclure LE, Love DN, Whalley JM (2000) EHV-1 glycoprotein D (EHV-1 gD) is required for virus entry and cell-cell fusion, and an EHV-1 gD deletion mutant induces a protective immune response in mice. Arch Virol 145:2371–2385

    Article  CAS  PubMed  Google Scholar 

  9. Foote CE, Love DN, Gilkerson JR, Rota J, Trevor-Jones P, Ruitenberg KM, Wellington JE, Whalley JM (2005) Serum antibody responses to equine herpesvirus 1 glycoprotein d in horses, pregnant mares and young foals. Vet Immunol Immunopathol 105:47–57

    Article  CAS  PubMed  Google Scholar 

  10. Weerasinghe CU, Learmonth GS, Gilkerson JR, Foote CE, Wellington JE, Whalley JM (2006) Equine herpesvirus 1 glycoprotein d expressed in E. coli provides partial protection against equine herpesvirus infection in mice and elicits virus-neutralizing antibodies in the horse. Vet Immunol Immunopathol 111:59–66

    Article  CAS  PubMed  Google Scholar 

  11. Ruitenberg KM, Walker C, Wellington JE, Love DN, Whalley JM (1999) Dna-mediated immunization with glycoprotein d of equine herpesvirus 1 (EHV-1) in a murine model of EHV-1 respiratory infection. Vaccine 17:237–244

    Article  CAS  PubMed  Google Scholar 

  12. Walker C, Ruitenberg KM, Love DN, Whalley JM (2000) Immunization of BALB/c mice with DNA encoding equine herpesvirus 1 (EHV-1) glycoprotein D affords partial protection in a model of EHV-1-induced abortion. Vet Microbiol 76:211–220

    Article  CAS  PubMed  Google Scholar 

  13. Minke JM, Fischer L, Baudu P, Guigal PM, Sindle T, Mumford JA, Audonnet JC (2006) Use of DNA and recombinant canarypox viral (ALVAC) vectors for equine herpes virus vaccination. Vet Immunol Immunopathol 111:47–57

    Article  CAS  PubMed  Google Scholar 

  14. Xiong D, Pan Z, Kang X, Wang J, Song L, Jiao X (2014) Molecular cloning and functional analysis of duck Toll-like receptor 5. Res Vet Sci 97:43–45

    Article  CAS  PubMed  Google Scholar 

  15. Mizel SB, Bates JT (2010) Flagellin as an adjuvant: cellular mechanisms and potential. J Immunol 185:5677–5682

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. St PM, Brisbin JT, Barjesteh N, Villaneueva AI, Parvizi P, Read LR, Nagy E, Sharif S (2014) Avian influenza virus vaccines containing toll-like receptors 2 and 5 ligand adjuvants promote protective immune responses in chickens. Viral Immunol 27:160–166

    Article  CAS  Google Scholar 

  17. Applequist SE, Rollman E, Wareing MD, Lidén M, Rozell B, Hinkula J, Ljunggren HG (2005) Activation of innate immunity, inflammation, and potentiation of DNA vaccination through mammalian expression of the TLR5 agonist flagellin. J Immunol 175:388–391

    Article  Google Scholar 

  18. Steinhagen F, Kinjo T, Bode C, Klinman DM (2011) Tlr-based immune adjuvants. Vaccine 29:3341–3355

    Article  CAS  PubMed  Google Scholar 

  19. Liu G, Tarbet B, Song L, Reiserova L, Weaver B, Chen Y, Li H, Hou F, Liu X, Parent J, Umlauf S, Shaw A, Tussey L (2011) Immunogenicity and efficacy of flagellin-fused vaccine candidates targeting 2009 pandemic H1N1 influenza in mice. PLoS One 6:e20928

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. 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:e2257

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Turley CB, Rupp RE, Johnson C, Taylor DN, Wolfson J, Tussey L, Kavita U, Stanberry L, Shaw A (2011) Safety and immunogenicity of a recombinant M2e-flagellin influenza vaccine (STF2.4xM2e) in healthy adults. Vaccine 29:5145–5152

    Article  CAS  PubMed  Google Scholar 

  22. Asadi-Karam MR, 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 31:1210–1216

    Article  CAS  PubMed  Google Scholar 

  23. Le MV, Robreau G, Mahana W (2008) Flagellin as a good carrier and potent adjuvant for Th1 response: study of mice immune response to the p27 (Rv2108) Mycobacterium tuberculosis antigen. Mol Immunol 45:2499–2507

    Article  CAS  Google Scholar 

  24. Yin G, Qin M, Liu X, Suo J, Tang X, Tao G, Han Q, Suo X, Wu W (2013) An Eimeria vaccine candidate based on Eimeria tenella immune mapped protein 1 and the TLR-5 agonist Salmonella typhimurium, FliC flagellin. Biochem Biophys Res Commun 440:437–442

    Article  CAS  PubMed  Google Scholar 

  25. Ramos-Sevillano E, Urzainqui A, Campuzano S, Moscoso M, González-Camacho F, Domenech M, Rodríguez de Córdoba S, Sánchez-Madrid F, Brown JS, García E, Yuste J (2015) Pleiotropic effects of cell wall amidase LytA on Streptococcus pneumoniae sensitivity to the host immune response. Infect Immun 83:591–603

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. 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 30:5761–5769

    Article  CAS  PubMed  Google Scholar 

  27. Delaney KN, Phipps JP, Johnson JB, Mizel SB (2010) A recombinant flagellin-poxvirus fusion protein vaccine elicits complement-dependent protection against respiratory challenge with vaccinia virus in mice. Viral Immunol 23:201–210

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Koh CY, Yuan D (2000) The functional relevance of NK-cell-mediated upregulation of antigen-specific IgG2a responses. Cell Immunol 204:135–142

    Article  CAS  PubMed  Google Scholar 

  29. Wang C, Li X, Wu T, Li D, Niu M, Wang Y, Zhang C, Cheng X, Chen P (2014) Bursin-like peptide (BLP) enhances H9N2 influenza vaccine induced humoral and cell mediated immune responses. Cell Immunol 292:57–64

    Article  CAS  PubMed  Google Scholar 

  30. Huber VC, Mckeon RM, Brackin MN, Miller LA, Keating R, Brown SA, Makarova N, Perez DR, Macdonald GH, McCullers JA (2006) Distinct contributions of vaccine-induced immunoglobulin G1 (IgG1) and IgG2a antibodies to protective immunity against Influenza. Clin Vaccine Immunol 13:981–990

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Song L, Xiong D, Kang X, Yang Y, Wang J, Guo Y, Xu H, Chen S, Peng D, Pan Z, Jiao X (2015) An avian influenza A (H7N9) virus vaccine candidate based on the fusion protein of hemagglutinin globular head and salmonella typhimurium, flagellin. BMC Biotechnol 15:1–8

    Article  CAS  Google Scholar 

  32. Dziadek B, Gatkowska J, Brzostek A, Dziadek J, Dzitko K, Grzybowski M, Dlugonska H (2011) Evaluation of three recombinant multi-antigenic vaccines composed of surface and secretory antigens of Toxoplasma gondii in murine models of experimental toxoplasmosis. Vaccine 29:821–830

    Article  CAS  PubMed  Google Scholar 

  33. Sack BK, Wang X, Sherman A, Rogers GL, Markusic DM (2015) Immune responses to human factor IX in haemophilia B mice of different genetic backgrounds are distinct and modified by TLR4. Haemophilia 21:133–139

    Article  CAS  PubMed  Google Scholar 

  34. Labarthe MC, Theocharous P, Russell N, Todryk S, Bangma C, Thraves P, Dalgleish AG, Whelan MA (2008) A novel murine model of allogeneic vaccination against prostate cancer. Cancer Immunol Immunother 57:453–465

    Article  PubMed  Google Scholar 

  35. Weimer ET, Lu H, Kock ND, Wozniak DJ, Mizel SB (2009) A fusion protein vaccine containing OprF epitope 8, OprI and type A and B flagellins promotes enhanced clearance of nonmucoid pseudomonas aeruginosa. Infect Immun 77:2356–2366

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Liu G, Song L, Reiserova L, Trivedi U, Li H, Liu X, Noah D, Hou F, Weaver B, Tussey L (2012) Flagellin-HA vaccines protect ferrets and mice against H5N1 highly pathogenic avian influenza virus (HPAIV) infections. Vaccine 30:6833–6838

    Article  CAS  PubMed  Google Scholar 

  37. Tewari D, Whalley JM, Love DN, Field HJ (1994) Characterization of immune responses to baculovirus-expressed equine herpesvirus type 1 glycoproteins D and H in a murine model. J Gen Virol 75:1735–1741

    Article  CAS  PubMed  Google Scholar 

  38. Zhang Y, Smith PM, Jennings SR, O’Callaghan DJ (2000) Quantitation of virus-specific classes of antibodies following immunization of mice with attenuated equine herpesvirus 1 and viral glycoprotein D. Virology 268:482–492

    Article  CAS  PubMed  Google Scholar 

  39. Ruitenberg KM, Love DN, Gilkerson JR, Wellington JE, Whalley JM (2000) Equine herpesvirus 1 (EHV-1) glycoprotein D DNA inoculation in horses with pre-existing EHV-1/EHV-4 antibody. Vet Microbiol 76:117–127

    Article  CAS  PubMed  Google Scholar 

  40. Ruitenberg KM, Walker C, Love DN, Wellington JE, Whalley JM (2000) A prime-boost immunization strategy with DNA and recombinant baculovirus-expressed protein enhances protective immunogenicity of glycoprotein D of equine herpesvirus 1 in naive and infection-primed mice. Vaccine 18:1367–1373

    Article  CAS  PubMed  Google Scholar 

  41. Allen GP, Kydd JH, Slater JD, Smith KC (1999) Recent advances in understanding the pathogenesis, latency and immunology of EHV-1 abortion (invited review). In: Proceedings of the 8th international conference on equine infectious diseases, Newmarket, pp 129–146

  42. Zeitlin L, Cone RA, Whaley KJ (1999) Using monoclonal antibodies to prevent mucosal transmission of epidemic infectious diseases. Emerg Infect Dis 5:54

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Israel BA, Herber R, Gao Y (1992) Induction of a mucosal barrier to bovine herpesvirus 1 replication in cattle. Virology 188:256–264

    Article  CAS  PubMed  Google Scholar 

  44. Breathnach CC, Yeargan MR, Sheoran AS, Allen GP (2001) The mucosal humoral immune response of the horse to infective challenge and vaccination with equine herpesvirus-1 antigens. Equine Vet J 33:651–657

    Article  CAS  PubMed  Google Scholar 

  45. Fuentealba NA, Zanuzzi CN, Scrochi MR, Sguazza GH, Bravi ME, Paz VCDL, Corva SG, Portiansky EL, Gimeno EJ, Barbeito CG, Galosi CM (2014) Protective effects of intranasal immunization with recombinant glycoprotein d in pregnant BALB/c mice challenged with different strains of equine herpesvirus 1. J Comp Pathol 151:384–393

    Article  CAS  PubMed  Google Scholar 

  46. Lee SE, Kim SY, Jeong BC, Kim YR, Bae SJ, Ahn OS, Lee JJ, Song HC, Kim JM, Choy HE, Chung SS, Kweon MN, Rhee JH (2006) A bacterial flagellin, vibrio vulnificus FlaB, has a strong mucosal adjuvant activity to induce protective immunity. Infect Immun 74:694–702

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Carapau D, Mitchell R, Nacer A, Shaw A, Othoro C, Frevert U, Nardin E (2013) Protective humoral immunity elicited by a needle-free malaria vaccine comprised of a chimeric plasmodium falciparum circumsporozoite protein and a Toll-like receptor 5 agonist, flagellin. Infect Immun 81:4350–4362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Author notes

  1. Yanan Zhao and Jianxin Chang equal contributors.

Authors and Affiliations

  1. Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Xinjiang Agricultural University, 311 Nongda East Road, Ürümqi, 830052, China

    Yanan Zhao, Jianxin Chang, Baojiang Zhang, Panpan Tong, Caidie Wang, Duoliang Ran & Yan Su

Authors
  1. Yanan Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jianxin Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Baojiang Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Panpan Tong
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Caidie Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Duoliang Ran
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yan Su
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yan Su.

Ethics declarations

Conflict of interest

The authors declare that they have no competing financial interests.

Ethical approval

All procedures involving animals were approved by the Ethics Committee for Animal Experiments at the Xinjiang Agricultural University. All animal experiments were conducted in strict accordance with international, national, and institutional guidelines for the use and care of animals. All authors contributed to this work and agreed to its publication.

Additional information

Handling Editor: Scott Schmid.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhao, Y., Chang, J., Zhang, B. et al. TLR-5 agonist Salmonella abortus equi flagellin FliC enhances FliC-gD-based DNA vaccination against equine herpesvirus 1 infection. Arch Virol 164, 1371–1382 (2019). https://doi.org/10.1007/s00705-019-04201-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-019-04201-4

Search

Navigation