Analyze Diet
Vaccine2012; 30(26); 3965-3974; doi: 10.1016/j.vaccine.2012.03.026

Immunogenicity and clinical protection against equine influenza by DNA vaccination of ponies.

Abstract: Equine influenza A (H3N8) virus infection is a leading cause of respiratory disease in horses, resulting in widespread morbidity and economic losses. As with influenza in other species, equine influenza strains continuously mutate, often requiring the development of new vaccines. Current inactivated (killed) vaccines, while efficacious, only offer limited protection against diverse subtypes and require frequent boosts. Research into new vaccine technologies, including gene-based vaccines, aims to increase the neutralization potency, breadth, and duration of protective immunity. Here, we demonstrate that a DNA vaccine expressing the hemagglutinin protein of equine H3N8 influenza virus generates homologous and heterologous immune responses, and protects against clinical disease and viral replication by homologous H3N8 virus in horses. Furthermore, we demonstrate that needle-free delivery is as efficient and effective as conventional parenteral injection using a needle and syringe. These findings suggest that DNA vaccines offer a safe, effective, and promising alternative approach for veterinary vaccines against equine influenza.
Publication Date: 2012-03-23 PubMed ID: 22449425PubMed Central: PMC3474358DOI: 10.1016/j.vaccine.2012.03.026Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
  • Journal Article
  • Research Support
  • N.I.H.
  • Intramural
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article examines the potential of a DNA vaccine in providing immunization against equine influenza A (H3N8) in horses. Results showed that this DNA vaccine, unlike inactivated (killed) vaccines, effectively generated both homologous and heterologous immune responses, offering significant protection from influenza symptoms and viral replication. Moreover, needle-free delivery proved just as efficient as traditional injection methods.

Equine Influenza and Vaccination Challenges

  • Equine influenza A, caused by the H3N8 virus subtype, is a severe respiratory disease affecting horses globally. This illness not only harms the equine population but also results in significant economic setbacks due to horse morbidity.
  • The constant mutation of these influenza strains often necessitates the formulation of new vaccines to ensure effective immunization.
  • The existing inactivated vaccines have their set of drawbacks. They provide short-term protection and are limited in their coverage against various subtypes. Therefore, regular vaccine boosters are required, which can be impractical and expensive.

New Vaccine Approach – Gene-based Vaccines

  • Researchers are exploring innovative vaccine technologies to overcome the limitations of inactivated vaccines. Gene-based or DNA vaccines are one such promising approach.
  • The researchers in this study developed a DNA vaccine that expresses the hemagglutinin protein of the H3N8 influenza virus. Hemagglutinin is a protein that allows the virus to enter the host’s cells, triggering an immune response.
  • The proposed DNA vaccine managed to generate both homologous responses (against identical strains of the infectious agent) and heterologous responses (against different strains), indicating a more significant protective immunization coverage.

Protective Immunity and Administration Methods

  • In trial conditions, the DNA vaccine successfully protected the test subjects from clinical influenza disease and minimized viral replication in those inflicted with a similar H3N8 virus strain.
  • The researchers also tested different delivery methods for the vaccine. They found that needle-free administration was equally efficient and effective as conventional needle injections. This opens up the potential for more humane and less stressful inoculation methods for horses.

Conclusions

  • The outcomes of this research suggest that DNA vaccines provide a safe, potent, and promising alternative to traditional inactivated vaccines in combating equine influenza.
  • This vaccine design not only offers broader protective immunity but also opens the door for exploring needle-free immunization techniques that could improve the overall vaccination experience for horses.

Cite This Article

APA
Ault A, Zajac AM, Kong WP, Gorres JP, Royals M, Wei CJ, Bao S, Yang ZY, Reedy SE, Sturgill TL, Page AE, Donofrio-Newman J, Adams AA, Balasuriya UB, Horohov DW, Chambers TM, Nabel GJ, Rao SS. (2012). Immunogenicity and clinical protection against equine influenza by DNA vaccination of ponies. Vaccine, 30(26), 3965-3974. https://doi.org/10.1016/j.vaccine.2012.03.026

Publication

ISSN: 1873-2518
NlmUniqueID: 8406899
Country: Netherlands
Language: English
Volume: 30
Issue: 26
Pages: 3965-3974

Researcher Affiliations

Ault, Alida
  • Vaccine Research Center (VRC), NIAID, NIH, Bethesda, MD 20892, USA.
Zajac, Alyse M
    Kong, Wing-Pui
      Gorres, J Patrick
        Royals, Michael
          Wei, Chih-Jen
            Bao, Saran
              Yang, Zhi-yong
                Reedy, Stephanie E
                  Sturgill, Tracy L
                    Page, Allen E
                      Donofrio-Newman, Jennifer
                        Adams, Amanda A
                          Balasuriya, Udeni B R
                            Horohov, David W
                              Chambers, Thomas M
                                Nabel, Gary J
                                  Rao, Srinivas S

                                    MeSH Terms

                                    • Animals
                                    • Antibodies, Viral / blood
                                    • Hemagglutinin Glycoproteins, Influenza Virus / genetics
                                    • Hemagglutinin Glycoproteins, Influenza Virus / immunology
                                    • Horse Diseases / prevention & control
                                    • Horses
                                    • Influenza A Virus, H3N8 Subtype / genetics
                                    • Influenza A Virus, H3N8 Subtype / immunology
                                    • Influenza Vaccines / administration & dosage
                                    • Influenza Vaccines / adverse effects
                                    • Influenza Vaccines / isolation & purification
                                    • Mice
                                    • Orthomyxoviridae Infections / prevention & control
                                    • Orthomyxoviridae Infections / veterinary
                                    • Vaccination / adverse effects
                                    • Vaccination / methods
                                    • Vaccines, DNA / administration & dosage
                                    • Vaccines, DNA / adverse effects
                                    • Vaccines, DNA / immunology

                                    Grant Funding

                                    • ZIG AI005117-01 / Intramural NIH HHS

                                    Conflict of Interest Statement

                                    To fully disclose any potential conflicts of interest, authors Gary J. Nabel, Srinivas S. Rao, Wing-Pui Kong, and Chih-Jen Wei are each listed on a patent filing for our DNA vaccine technology, entitled “U.S. Continuation in Part Patent Application No. 12/838,292”, which is an adjunct to an existing patent entitled “Influenza DNA Vaccination and Methods of Use thereof”, Serial #61/023,341.

                                    References

                                    This article includes 64 references
                                    1. Aerts JL, Gonzales MI, Topalian SL. Selection of appropriate control genes to assess expression of tumor antigens using real-time RT-PCR.. Biotechniques 2004 Jan;36(1):84-6, 88, 90-1.
                                      pubmed: 14740490doi: 10.2144/04361st04google scholar: lookup
                                    2. Amorij JP, Hinrichs WLj, Frijlink HW, Wilschut JC, Huckriede A. Needle-free influenza vaccination.. Lancet Infect Dis 2010 Oct;10(10):699-711.
                                      pubmed: 20883966doi: 10.1016/s1473-3099(10)70157-2google scholar: lookup
                                    3. Barouch DH, Yang ZY, Kong WP, Korioth-Schmitz B, Sumida SM, Truitt DM, Kishko MG, Arthur JC, Miura A, Mascola JR, Letvin NL, Nabel GJ. A human T-cell leukemia virus type 1 regulatory element enhances the immunogenicity of human immunodeficiency virus type 1 DNA vaccines in mice and nonhuman primates.. J Virol 2005 Jul;79(14):8828-34.
                                    4. Boliar S, Stanislawek W, Chambers TM. Inability of kaolin treatment to remove nonspecific inhibitors from equine serum for the hemagglutination inhibition test against equine H7N7 influenza virus.. J Vet Diagn Invest 2006 May;18(3):264-7.
                                      pubmed: 16789714doi: 10.1177/104063870601800305google scholar: lookup
                                    5. Breathnach CC, Clark HJ, Clark RC, Olsen CW, Townsend HG, Lunn DP. Immunization with recombinant modified vaccinia Ankara (rMVA) constructs encoding the HA or NP gene protects ponies from equine influenza virus challenge.. Vaccine 2006 Feb 20;24(8):1180-90.
                                      pubmed: 16194586doi: 10.1016/j.vaccine.2005.08.091google scholar: lookup
                                    6. Bryant NA, Rash AS, Russell CA, Ross J, Cooke A, Bowman S, MacRae S, Lewis NS, Paillot R, Zanoni R, Meier H, Griffiths LA, Daly JM, Tiwari A, Chambers TM, Newton JR, Elton DM. Antigenic and genetic variations in European and North American equine influenza virus strains (H3N8) isolated from 2006 to 2007.. Vet Microbiol 2009 Jul 2;138(1-2):41-52.
                                      pubmed: 19346084doi: 10.1016/j.vetmic.2009.03.004google scholar: lookup
                                    7. Bryant NA, Rash AS, Woodward AL, Medcalf E, Helwegen M, Wohlfender F, Cruz F, Herrmann C, Borchers K, Tiwari A, Chambers TM, Newton JR, Mumford JA, Elton DM. Isolation and characterisation of equine influenza viruses (H3N8) from Europe and North America from 2008 to 2009.. Vet Microbiol 2011 Jan 10;147(1-2):19-27.
                                      pubmed: 20580170doi: 10.1016/j.vetmic.2010.05.040google scholar: lookup
                                    8. Chambers TM, Holland RE, Tudor LR, Townsend HG, Cook A, Bogdan J, Lunn DP, Hussey S, Whitaker-Dowling P, Youngner JS, Sebring RW, Penner SJ, Stiegler GL. A new modified live equine influenza virus vaccine: phenotypic stability, restricted spread and efficacy against heterologous virus challenge.. Equine Vet J 2001 Nov;33(7):630-6.
                                      pubmed: 11770982doi: 10.2746/042516401776249291google scholar: lookup
                                    9. Chambers TM, Quinlivan M, Sturgill T, Cullinane A, Horohov DW, Zamarin D, Arkins S, García-Sastre A, Palese P. Influenza A viruses with truncated NS1 as modified live virus vaccines: pilot studies of safety and efficacy in horses.. Equine Vet J 2009 Jan;41(1):87-92.
                                      pmc: PMC2878972pubmed: 19301588doi: 10.2746/042516408x371937google scholar: lookup
                                    10. Crouch CF, Daly J, Henley W, Hannant D, Wilkins J, Francis MJ. The use of a systemic prime/mucosal boost strategy with an equine influenza ISCOM vaccine to induce protective immunity in horses.. Vet Immunol Immunopathol 2005 Dec 15;108(3-4):345-55.
                                      pubmed: 16098611doi: 10.1016/j.vetimm.2005.06.009google scholar: lookup
                                    11. Cullinane A, Elton D, Mumford J. Equine influenza - surveillance and control.. Influenza Other Respir Viruses 2010 Nov;4(6):339-44.
                                    12. Daly JM, MacRae S, Newton JR, Wattrang E, Elton DM. Equine influenza: a review of an unpredictable virus.. Vet J 2011 Jul;189(1):7-14.
                                      pubmed: 20685140doi: 10.1016/j.tvjl.2010.06.026google scholar: lookup
                                    13. Daly JM, Newton JR, Mumford JA. Current perspectives on control of equine influenza.. Vet Res 2004 Jul-Aug;35(4):411-23.
                                      pubmed: 15236674doi: 10.1051/vetres:2004023google scholar: lookup
                                    14. Daly JM, Sindle T, Tearle J, Barquero N, Newton JR, Corning S. Equine influenza vaccine containing older H3N8 strains offers protection against A/eq/South Africa/4/03 (H3N8) strain in a short-term vaccine efficacy study.. Equine Vet J 2007 Sep;39(5):446-50.
                                      pubmed: 17910270doi: 10.2746/042516407x180327google scholar: lookup
                                    15. Dheda K, Huggett JF, Bustin SA, Johnson MA, Rook G, Zumla A. Validation of housekeeping genes for normalizing RNA expression in real-time PCR.. Biotechniques 2004 Jul;37(1):112-4, 116, 118-9.
                                      pubmed: 15283208doi: 10.2144/04371rr03google scholar: lookup
                                    16. van Drunen Littel-van den Hurk S, Babiuk S, Babiuk LA. Needle-free delivery of veterinary DNA vaccines.. Methods Mol Med 2006;127:91-105.
                                      pubmed: 16988449doi: 10.1385/1-59745-168-1:91google scholar: lookup
                                    17. Edlund Toulemonde C, Daly J, Sindle T, Guigal PM, Audonnet JC, Minke JM. Efficacy of a recombinant equine influenza vaccine against challenge with an American lineage H3N8 influenza virus responsible for the 2003 outbreak in the United Kingdom.. Vet Rec 2005 Mar 19;156(12):367-71.
                                      pubmed: 15816180doi: 10.1136/vr.156.12.367google scholar: lookup
                                    18. Ekwueme DU, Weniger BG, Chen RT. Model-based estimates of risks of disease transmission and economic costs of seven injection devices in sub-Saharan Africa.. Bull World Health Organ 2002;80(11):859-70.
                                      pmc: PMC2567682pubmed: 12481207
                                    19. Feltquate DM, Heaney S, Webster RG, Robinson HL. Different T helper cell types and antibody isotypes generated by saline and gene gun DNA immunization.. J Immunol 1997 Mar 1;158(5):2278-84.
                                      pubmed: 9036975
                                    20. Gorres JP, Lager KM, Kong WP, Royals M, Todd JP, Vincent AL, Wei CJ, Loving CL, Zanella EL, Janke B, Kehrli ME Jr, Nabel GJ, Rao SS. DNA vaccination elicits protective immune responses against pandemic and classic swine influenza viruses in pigs.. Clin Vaccine Immunol 2011 Nov;18(11):1987-95.
                                      pmc: PMC3209026pubmed: 21918118doi: 10.1128/cvi.05171-11google scholar: lookup
                                    21. Heldens JG, Pouwels HG, Derks CG, Van de Zande SM, Hoeijmakers MJ. The first safe inactivated equine influenza vaccine formulation adjuvanted with ISCOM-Matrix that closes the immunity gap.. Vaccine 2009 Sep 4;27(40):5530-7.
                                      pubmed: 19607950doi: 10.1016/j.vaccine.2009.06.085google scholar: lookup
                                    22. Heldens JG, Pouwels HG, Derks CG, Van de Zande SM, Hoeijmakers MJ. Duration of immunity induced by an equine influenza and tetanus combination vaccine formulation adjuvanted with ISCOM-Matrix.. Vaccine 2010 Oct 8;28(43):6989-96.
                                      pubmed: 20728523doi: 10.1016/j.vaccine.2010.08.043google scholar: lookup
                                    23. Huber VC, McKeon RM, Brackin MN, Miller LA, Keating R, Brown SA, Makarova N, Perez DR, Macdonald GH, McCullers JA. Distinct contributions of vaccine-induced immunoglobulin G1 (IgG1) and IgG2a antibodies to protective immunity against influenza.. Clin Vaccine Immunol 2006 Sep;13(9):981-90.
                                      pmc: PMC1563571pubmed: 16960108doi: 10.1128/cvi.00156-06google scholar: lookup
                                    24. Huber VC, Thomas PG, McCullers JA. A multi-valent vaccine approach that elicits broad immunity within an influenza subtype.. Vaccine 2009 Feb 18;27(8):1192-200.
                                    25. Johnson PA, Conway M, Daly J, Nicolson C, Robertson JS, Mills KHG. Influenza HA DNA induces Th1 cells and protection despite limited antibody responses.. Int Congr Ser 2001;1219:911–915.
                                    26. Kannegieter NJ, Frogley A, Crispe E, Kirkland PD. Clinical outcomes and virology of equine influenza in a naïve population and in horses infected soon after receiving one dose of vaccine.. Aust Vet J 2011 Jul;89 Suppl 1:139-42.
                                    27. Kong WP, Hood C, Yang ZY, Wei CJ, Xu L, García-Sastre A, Tumpey TM, Nabel GJ. Protective immunity to lethal challenge of the 1918 pandemic influenza virus by vaccination.. Proc Natl Acad Sci U S A 2006 Oct 24;103(43):15987-91.
                                      pmc: PMC1613227pubmed: 17043214doi: 10.1073/pnas.0607564103google scholar: lookup
                                    28. Kutzler MA, Weiner DB. DNA vaccines: ready for prime time?. Nat Rev Genet 2008 Oct;9(10):776-88.
                                      pmc: PMC4317294pubmed: 18781156doi: 10.1038/nrg2432google scholar: lookup
                                    29. Letvin NL, Rao SS, Montefiori DC, Seaman MS, Sun Y, Lim SY, Yeh WW, Asmal M, Gelman RS, Shen L, Whitney JB, Seoighe C, Lacerda M, Keating S, Norris PJ, Hudgens MG, Gilbert PB, Buzby AP, Mach LV, Zhang J, Balachandran H, Shaw GM, Schmidt SD, Todd JP, Dodson A, Mascola JR, Nabel GJ. Immune and Genetic Correlates of Vaccine Protection Against Mucosal Infection by SIV in Monkeys.. Sci Transl Med 2011 May 4;3(81):81ra36.
                                    30. Lewis NS, Daly JM, Russell CA, Horton DL, Skepner E, Bryant NA, Burke DF, Rash AS, Wood JL, Chambers TM, Fouchier RA, Mumford JA, Elton DM, Smith DJ. Antigenic and genetic evolution of equine influenza A (H3N8) virus from 1968 to 2007.. J Virol 2011 Dec;85(23):12742-9.
                                      pmc: PMC3209411pubmed: 21937642doi: 10.1128/jvi.05319-11google scholar: lookup
                                    31. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.. Methods 2001 Dec;25(4):402-8.
                                      pubmed: 11846609doi: 10.1006/meth.2001.1262google scholar: lookup
                                    32. Lopez AM, Hecker R, Mutwiri G, van Drunen Littel-van den Hurk S, Babiuk LA, Townsend HG. Formulation with CpG ODN enhances antibody responses to an equine influenza virus vaccine.. Vet Immunol Immunopathol 2006 Nov 15;114(1-2):103-10.
                                      pubmed: 16950519doi: 10.1016/j.vetimm.2006.07.013google scholar: lookup
                                    33. Lu Z, Chambers TM, Boliar S, Branscum AJ, Sturgill TL, Timoney PJ, Reedy SE, Tudor LR, Dubovi EJ, Vickers ML, Sells S, Balasuriya UB. Development and evaluation of one-step TaqMan real-time reverse transcription-PCR assays targeting nucleoprotein, matrix, and hemagglutinin genes of equine influenza virus.. J Clin Microbiol 2009 Dec;47(12):3907-13.
                                      pmc: PMC2786649pubmed: 19846644doi: 10.1128/jcm.00598-09google scholar: lookup
                                    34. Lunn DP, Hussey S, Sebing R, Rushlow KE, Radecki SV, Whitaker-Dowling P, Youngner JS, Chambers TM, Holland RE Jr, Horohov DW. Safety, efficacy, and immunogenicity of a modified-live equine influenza virus vaccine in ponies after induction of exercise-induced immunosuppression.. J Am Vet Med Assoc 2001 Mar 15;218(6):900-6.
                                      pubmed: 11294315doi: 10.2460/javma.2001.218.900google scholar: lookup
                                    35. Lunn DP, Soboll G, Schram BR, Quass J, McGregor MW, Drape RJ, Macklin MD, McCabe DE, Swain WF, Olsen CW. Antibody responses to DNA vaccination of horses using the influenza virus hemagglutinin gene.. Vaccine 1999 May 4;17(18):2245-58.
                                      pubmed: 10403592doi: 10.1016/s0264-410x(98)00496-4google scholar: lookup
                                    36. Minke JM, Toulemonde CE, Coupier H, Guigal PM, Dinic S, Sindle T, Jessett D, Black L, Bublot M, Pardo MC, Audonnet JC. Efficacy of a canarypox-vectored recombinant vaccine expressing the hemagglutinin gene of equine influenza H3N8 virus in the protection of ponies from viral challenge.. Am J Vet Res 2007 Feb;68(2):213-9.
                                      pubmed: 17269889doi: 10.2460/ajvr.68.2.213google scholar: lookup
                                    37. Morley PS, Townsend HG, Bogdan JR, Haines DM. Efficacy of a commercial vaccine for preventing disease caused by influenza virus infection in horses.. J Am Vet Med Assoc 1999 Jul 1;215(1):61-6.
                                      pubmed: 10397067
                                    38. Mumford J, Wood JM, Scott AM, Folkers C, Schild GC. Studies with inactivated equine influenza vaccine. 2. Protection against experimental infection with influenza virus A/equine/Newmarket/79 (H3N8).. J Hyg (Lond) 1983 Jun;90(3):385-95.
                                      pmc: PMC2134271pubmed: 6306098doi: 10.1017/s0022172400029016google scholar: lookup
                                    39. Mumford JA, Hannant D, Jessett DM. Experimental infection of ponies with equine influenza (H3N8) viruses by intranasal inoculation or exposure to aerosols.. Equine Vet J 1990 Mar;22(2):93-8.
                                    40. Mumford JA, Wood JM, Folkers C, Schild GC. Protection against experimental infection with influenza virus A/equine/Miami/63 (H3N8) provided by inactivated whole virus vaccines containing homologous virus.. Epidemiol Infect 1988 Jun;100(3):501-10.
                                      pmc: PMC2249356pubmed: 2837409doi: 10.1017/s0950268800067236google scholar: lookup
                                    41. Nelson KM, Schram BR, McGregor MW, Sheoran AS, Olsen CW, Lunn DP. Local and systemic isotype-specific antibody responses to equine influenza virus infection versus conventional vaccination.. Vaccine 1998 Aug;16(13):1306-13.
                                      pubmed: 9682395doi: 10.1016/s0264-410x(98)00009-7google scholar: lookup
                                    42. Newton JR, Townsend HG, Wood JL, Sinclair R, Hannant D, Mumford JA. Immunity to equine influenza: relationship of vaccine-induced antibody in young Thoroughbred racehorses to protection against field infection with influenza A/equine-2 viruses (H3N8).. Equine Vet J 2000 Jan;32(1):65-74.
                                      pubmed: 10661388doi: 10.2746/042516400777612116google scholar: lookup
                                    43. OIE. OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Paris: Office International des Epizooties; 2004. Equine Influenza; pp. 686–697.
                                    44. Paillot R, Hannant D, Kydd JH, Daly JM. Vaccination against equine influenza: quid novi?. Vaccine 2006 May 8;24(19):4047-61.
                                      pubmed: 16545507doi: 10.1016/j.vaccine.2006.02.030google scholar: lookup
                                    45. Paillot R, Kydd JH, Sindle T, Hannant D, Edlund Toulemonde C, Audonnet JC, Minke JM, Daly JM. Antibody and IFN-gamma responses induced by a recombinant canarypox vaccine and challenge infection with equine influenza virus.. Vet Immunol Immunopathol 2006 Aug 15;112(3-4):225-33.
                                      pubmed: 16621023doi: 10.1016/j.vetimm.2006.02.007google scholar: lookup
                                    46. Quinlivan M, Nelly M, Prendergast M, Breathnach C, Horohov D, Arkins S, Chiang YW, Chu HJ, Ng T, Cullinane A. Pro-inflammatory and antiviral cytokine expression in vaccinated and unvaccinated horses exposed to equine influenza virus.. Vaccine 2007 Oct 10;25(41):7056-64.
                                      pubmed: 17825959doi: 10.1016/j.vaccine.2007.07.059google scholar: lookup
                                    47. Rainen L, Oelmueller U, Jurgensen S, Wyrich R, Ballas C, Schram J, Herdman C, Bankaitis-Davis D, Nicholls N, Trollinger D, Tryon V. Stabilization of mRNA expression in whole blood samples.. Clin Chem 2002 Nov;48(11):1883-90.
                                      pubmed: 12406972
                                    48. Rao S, Kong WP, Wei CJ, Yang ZY, Nason M, Styles D, DeTolla LJ, Panda A, Sorrell EM, Song H, Wan H, Ramirez-Nieto GC, Perez D, Nabel GJ. Multivalent HA DNA vaccination protects against highly pathogenic H5N1 avian influenza infection in chickens and mice.. PLoS One 2008;3(6):e2432.
                                    49. Rao SS, Styles D, Kong W, Andrews C, Gorres JP, Nabel GJ. A gene-based avian influenza vaccine in poultry.. Poult Sci 2009 Apr;88(4):860-6.
                                      pmc: PMC7194532pubmed: 19276436doi: 10.3382/ps.2008-00360google scholar: lookup
                                    50. Reed LJ, Muench H. A simple method of estimating fifty per cent endpoints.. Am. J. Epidemiol. 1938;27:493–497.
                                    51. Shipley JM, Miller RD, Wu BM, Grubb JH, Christensen SG, Kyle JW, Sly WS. Analysis of the 5' flanking region of the human beta-glucuronidase gene.. Genomics 1991 Aug;10(4):1009-18.
                                      pubmed: 1916806doi: 10.1016/0888-7543(91)90192-hgoogle scholar: lookup
                                    52. Shu Y, Winfrey S, Yang ZY, Xu L, Rao SS, Srivastava I, Barnett SW, Nabel GJ, Mascola JR. Efficient protein boosting after plasmid DNA or recombinant adenovirus immunization with HIV-1 vaccine constructs.. Vaccine 2007 Feb 9;25(8):1398-408.
                                    53. Soboll G, Horohov DW, Aldridge BM, Olsen CW, McGregor MW, Drape RJ, Macklin MD, Swain WF, Lunn DP. Regional antibody and cellular immune responses to equine influenza virus infection, and particle mediated DNA vaccination.. Vet Immunol Immunopathol 2003 Jul 15;94(1-2):47-62.
                                      pubmed: 12842611doi: 10.1016/s0165-2427(03)00060-6google scholar: lookup
                                    54. Soboll G, Hussey SB, Minke JM, Landolt GA, Hunter JS, Jagannatha S, Lunn DP. Onset and duration of immunity to equine influenza virus resulting from canarypox-vectored (ALVAC) vaccination.. Vet Immunol Immunopathol 2010 May 15;135(1-2):100-107.
                                      pubmed: 20018384doi: 10.1016/j.vetimm.2009.11.007google scholar: lookup
                                    55. Soboll G, Nelson KM, Leuthner ES, Clark RJ, Drape R, Macklin MD, Swain WF, Olsen CW, Lunn DP. Mucosal co-administration of cholera toxin and influenza virus hemagglutinin-DNA in ponies generates a local IgA response.. Vaccine 2003 Jun 20;21(21-22):3081-92.
                                      pubmed: 12798652doi: 10.1016/s0264-410x(03)00161-0google scholar: lookup
                                    56. Song K, Bolton DL, Wei CJ, Wilson RL, Camp JV, Bao S, Mattapallil JJ, Herzenberg LA, Herzenberg LA, Andrews CA, Sadoff JC, Goudsmit J, Pau MG, Seder RA, Kozlowski PA, Nabel GJ, Roederer M, Rao SS. Genetic immunization in the lung induces potent local and systemic immune responses.. Proc Natl Acad Sci U S A 2010 Dec 21;107(51):22213-8.
                                      pmc: PMC3009829pubmed: 21135247doi: 10.1073/pnas.1015536108google scholar: lookup
                                    57. Townsend HG, Penner SJ, Watts TC, Cook A, Bogdan J, Haines DM, Griffin S, Chambers T, Holland RE, Whitaker-Dowling P, Youngner JS, Sebring RW. Efficacy of a cold-adapted, intranasal, equine influenza vaccine: challenge trials.. Equine Vet J 2001 Nov;33(7):637-43.
                                      pubmed: 11770983doi: 10.2746/042516401776249354google scholar: lookup
                                    58. United States Department of Agriculture. USDA issues license for West Nile Virus DNA vaccine for horses. 2005 http://www. aphis. usda. gov/lpa/news/2005/07/wnvdna_vs. html. [Online.].
                                    59. Van de Walle GR, May MA, Peters ST, Metzger SM, Rosas CT, Osterrieder N. A vectored equine herpesvirus type 1 (EHV-1) vaccine elicits protective immune responses against EHV-1 and H3N8 equine influenza virus.. Vaccine 2010 Jan 22;28(4):1048-55.
                                      pubmed: 19897066doi: 10.1016/j.vaccine.2009.10.123google scholar: lookup
                                    60. van Maanen C, Cullinane A. Equine influenza virus infections: an update.. Vet Q 2002 Jun;24(2):79-94.
                                      pubmed: 12095083doi: 10.1080/01652176.2002.9695127google scholar: lookup
                                    61. Wattrang E, Jessett DM, Yates P, Fuxler L, Hannant D. Experimental infection of ponies with equine influenza A2 (H3N8) virus strains of different pathogenicity elicits varying interferon and interleukin-6 responses.. Viral Immunol 2003;16(1):57-67.
                                      pubmed: 12725689doi: 10.1089/088282403763635456google scholar: lookup
                                    62. Wei CJ, Boyington JC, McTamney PM, Kong WP, Pearce MB, Xu L, Andersen H, Rao S, Tumpey TM, Yang ZY, Nabel GJ. Induction of broadly neutralizing H1N1 influenza antibodies by vaccination.. Science 2010 Aug 27;329(5995):1060-4.
                                      pubmed: 20647428doi: 10.1126/science.1192517google scholar: lookup
                                    63. Wood JM, Mumford J, Folkers C, Scott AM, Schild GC. Studies with inactivated equine influenza vaccine. 1. Serological responses of ponies to graded doses of vaccine.. J Hyg (Lond) 1983 Jun;90(3):371-84.
                                      pmc: PMC2134282pubmed: 6345659doi: 10.1017/s0022172400029004google scholar: lookup
                                    64. Wright PF. Vaccine preparedness--are we ready for the next influenza pandemic?. N Engl J Med 2008 Jun 12;358(24):2540-3.
                                      pubmed: 18550873doi: 10.1056/nejmp0803650google scholar: lookup

                                    Citations

                                    This article has been cited 13 times.
                                    1. Whitlock F, Murcia PR, Newton JR. A Review on Equine Influenza from a Human Influenza Perspective.. Viruses 2022 Jun 15;14(6).
                                      doi: 10.3390/v14061312pubmed: 35746783google scholar: lookup
                                    2. Redmann RK, Kaushal D, Golden N, Threeton B, Killeen SZ, Kuehl PJ, Roy CJ. Particle Dynamics and Bioaerosol Viability of Aerosolized Bacillus Calmette-Guérin Vaccine Using Jet and Vibrating Mesh Clinical Nebulizers.. J Aerosol Med Pulm Drug Deliv 2022 Jan;35(1):50-56.
                                      doi: 10.1089/jamp.2021.0030pubmed: 34619040google scholar: lookup
                                    3. Oladunni FS, Oseni SO, Martinez-Sobrido L, Chambers TM. Equine Influenza Virus and Vaccines.. Viruses 2021 Aug 20;13(8).
                                      doi: 10.3390/v13081657pubmed: 34452521google scholar: lookup
                                    4. Chang C, Sun J, Hayashi H, Suzuki A, Sakaguchi Y, Miyazaki H, Nishikawa T, Nakagami H, Yamashita K, Kaneda Y. Stable Immune Response Induced by Intradermal DNA Vaccination by a Novel Needleless Pyro-Drive Jet Injector.. AAPS PharmSciTech 2019 Dec 9;21(1):19.
                                      doi: 10.1208/s12249-019-1564-zpubmed: 31820256google scholar: lookup
                                    5. Blanco-Lobo P, Rodriguez L, Reedy S, Oladunni FS, Nogales A, Murcia PR, Chambers TM, Martinez-Sobrido L. A Bivalent Live-Attenuated Vaccine for the Prevention of Equine Influenza Virus.. Viruses 2019 Oct 11;11(10).
                                      doi: 10.3390/v11100933pubmed: 31614538google scholar: lookup
                                    6. Singh RK, Dhama K, Karthik K, Khandia R, Munjal A, Khurana SK, Chakraborty S, Malik YS, Virmani N, Singh R, Tripathi BN, Munir M, van der Kolk JH. A Comprehensive Review on Equine Influenza Virus: Etiology, Epidemiology, Pathobiology, Advances in Developing Diagnostics, Vaccines, and Control Strategies.. Front Microbiol 2018;9:1941.
                                      doi: 10.3389/fmicb.2018.01941pubmed: 30237788google scholar: lookup
                                    7. Hinkula J, Petkov S, Ljungberg K, Hallengärd D, Bråve A, Isaguliants M, Falkeborn T, Sharma S, Liakina V, Robb M, Eller M, Moss B, Biberfeld G, Sandström E, Nilsson C, Markland K, Blomberg P, Wahren B. HIVIS-DNA or HIVISopt-DNA priming followed by CMDR vaccinia-based boosts induce both humoral and cellular murine immune responses to HIV.. Heliyon 2017 Jun;3(6):e00339.
                                      doi: 10.1016/j.heliyon.2017.e00339pubmed: 28721397google scholar: lookup
                                    8. Stachyra A, Redkiewicz P, Kosson P, Protasiuk A, Góra-Sochacka A, Kudla G, Sirko A. Codon optimization of antigen coding sequences improves the immune potential of DNA vaccines against avian influenza virus H5N1 in mice and chickens.. Virol J 2016 Aug 26;13(1):143.
                                      doi: 10.1186/s12985-016-0599-ypubmed: 27562235google scholar: lookup
                                    9. Borggren M, Nielsen J, Karlsson I, Dalgaard TS, Trebbien R, Williams JA, Fomsgaard A. A polyvalent influenza DNA vaccine applied by needle-free intradermal delivery induces cross-reactive humoral and cellular immune responses in pigs.. Vaccine 2016 Jul 12;34(32):3634-40.
                                      doi: 10.1016/j.vaccine.2016.05.030pubmed: 27211039google scholar: lookup
                                    10. Paillot R. A Systematic Review of Recent Advances in Equine Influenza Vaccination.. Vaccines (Basel) 2014 Nov 14;2(4):797-831.
                                      doi: 10.3390/vaccines2040797pubmed: 26344892google scholar: lookup
                                    11. Williams JA. Vector Design for Improved DNA Vaccine Efficacy, Safety and Production.. Vaccines (Basel) 2013 Jun 25;1(3):225-49.
                                      doi: 10.3390/vaccines1030225pubmed: 26344110google scholar: lookup
                                    12. Borggren M, Nielsen J, Bragstad K, Karlsson I, Krog JS, Williams JA, Fomsgaard A. Vector optimization and needle-free intradermal application of a broadly protective polyvalent influenza A DNA vaccine for pigs and humans.. Hum Vaccin Immunother 2015;11(8):1983-90.
                                      doi: 10.1080/21645515.2015.1011987pubmed: 25746201google scholar: lookup
                                    13. Stack JC, Murcia PR, Grenfell BT, Wood JL, Holmes EC. Inferring the inter-host transmission of influenza A virus using patterns of intra-host genetic variation.. Proc Biol Sci 2013 Jan 7;280(1750):20122173.
                                      doi: 10.1098/rspb.2012.2173pubmed: 23135678google scholar: lookup