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Home / Equine Research Bank / PLoS One / Immunogenic profile of a plant-produced nonavalent African h...
PloS one2024; 19(4); e0301340; doi: 10.1371/journal.pone.0301340

Immunogenic profile of a plant-produced nonavalent African horse sickness viral protein 2 (VP2) vaccine in IFNAR-/- mice.

Abstract: A safe, highly immunogenic multivalent vaccine to protect against all nine serotypes of African horse sickness virus (AHSV), will revolutionise the AHS vaccine industry in endemic countries and beyond. Plant-produced AHS virus-like particles (VLPs) and soluble viral protein 2 (VP2) vaccine candidates were developed that have the potential to protect against all nine serotypes but can equally well be formulated as mono- and bi-valent formulations for localised outbreaks of specific serotypes. In the first interferon α/β receptor knock-out (IFNAR-/-) mice trial conducted, a nine-serotype (nonavalent) vaccine administered as two pentavalent (5 μg per serotype) vaccines (VLP/VP2 combination or exclusively VP2), were directly compared to the commercially available AHS live attenuated vaccine. In a follow up trial, mice were vaccinated with an adjuvanted nine-serotype multivalent VP2 vaccine in a prime boost strategy and resulted in the desired neutralising antibody titres of 1:320, previously demonstrated to confer protective immunity in IFNAR-/- mice. In addition, the plant-produced VP2 vaccine performed favourably when compared to the commercial vaccine. Here we provide compelling data for a nonavalent VP2-based vaccine candidate, with the VP2 from each serotype being antigenically distinguishable based on LC-MS/MS and ELISA data. This is the first preclinical trial demonstrating the ability of an adjuvanted nonavalent cocktail of soluble, plant-expressed AHS VP2 proteins administered in a prime-boost strategy eliciting high antibody titres against all 9 AHSV serotypes. Furthermore, elevated T helper cells 2 (Th2) and Th1, indicative of humoral and cell-mediated memory T cell immune responses, respectively, were detected in mouse serum collected 14 days after the multivalent prime-boost vaccination. Both Th2 and Th1 may play a role to confer protective immunity. These preclinical immunogenicity studies paved the way to test the safety and protective efficacy of the plant-produced nonavalent VP2 vaccine candidate in the target animals, horses.
Copyright: © 2024 O’Kennedy et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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Publication Date: 2024-04-16 PubMed ID: 38625924PubMed Central: PMC11020708DOI: 10.1371/journal.pone.0301340Google Scholar: Lookup
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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.

Overview

  • This study evaluated a novel, plant-produced vaccine targeting all nine serotypes of African horse sickness virus (AHSV) in a mouse model lacking interferon α/β receptors (IFNAR-/- mice).
  • The vaccine, based on the viral protein 2 (VP2) from each serotype, showed strong immune responses and compared favorably with the existing live attenuated vaccine.

Background and Importance

  • African horse sickness (AHS) is a severe viral disease affecting horses, caused by African horse sickness virus (AHSV) that has nine distinct serotypes.
  • Effective vaccination against all nine serotypes is essential for comprehensive disease control in endemic regions and areas at risk of outbreaks.
  • Existing vaccines include live attenuated formulations but have limitations related to safety and logistical issues.
  • Developing a safe, multivalent vaccine that protects against all serotypes could revolutionize AHS control.
  • Plant-produced vaccines provide a promising alternative due to potential for rapid, scalable, and cost-effective production.

Vaccine Design and Production

  • The vaccine candidates focused on the viral protein 2 (VP2), a key immunogenic protein of AHSV responsible for eliciting neutralizing antibodies.
  • Two vaccine formats were produced in plants:
    • Virus-like particles (VLPs) incorporating VP2
    • Soluble VP2 proteins alone
  • These could be used as a multivalent (nonavalent) vaccine covering all nine serotypes or formulated as mono- or bi-valent vaccines for specific outbreak responses.

Experimental Model and Vaccination Strategy

  • The model used IFNAR-/- mice, which lack interferon α/β receptors, making them more susceptible to viral infections and thus a useful system for vaccine efficacy evaluation.
  • Two pentavalent vaccines (each including 5 μg per serotype) were administered:
    • One combining VLPs and VP2 proteins
    • One consisting exclusively of VP2 proteins
  • These were directly compared with the commercial live attenuated AHS vaccine in the first trial.
  • In a follow-up trial, the soluble VP2 nonavalent vaccine was tested using an adjuvanted, prime-boost approach to enhance immune responses.

Key Findings

  • Both vaccine formulations induced strong neutralizing antibody titers against all nine AHSV serotypes; the follow-up vaccine produced titers of 1:320, a level known to confer protection in this mouse model.
  • Plant-produced VP2 vaccine results were comparable or favorable compared to the commercial live attenuated vaccine.
  • LC-MS/MS and ELISA assays confirmed that VP2 proteins from each serotype retained antigenic distinctness when combined into the nonavalent vaccine, indicating broad coverage.
  • Immune profiling showed increased T helper cell responses of both Th1 and Th2 types:
    • Th2 cells suggest a strong humoral (antibody) memory response
    • Th1 cells indicate a cell-mediated immune memory response
  • Both Th1 and Th2 responses may be important for providing protective immunity against AHSV.

Implications and Future Directions

  • This is the first preclinical demonstration that a plant-expressed, adjuvanted, nonavalent VP2 protein vaccine given by prime-boost can induce cross-serotype immune responses suitable for protection.
  • The data support advancing this vaccine candidate to safety and efficacy trials in the natural target species, horses.
  • Plant-based production could enable scalable, safe, and effective vaccines aiding global AHS control efforts, especially in endemic countries.

Cite This Article

APA
O'Kennedy MM, Roth R, Ebersohn K, du Plessis LH, Mamputha S, Rutkowska DA, du Preez I, Verschoor JA, Lemmer Y. (2024). Immunogenic profile of a plant-produced nonavalent African horse sickness viral protein 2 (VP2) vaccine in IFNAR-/- mice. PLoS One, 19(4), e0301340. https://doi.org/10.1371/journal.pone.0301340

Publication

PloS one
ISSN: 1932-6203
NlmUniqueID: 101285081
Country: United States
Language: English
Volume: 19
Issue: 4
Pages: e0301340
PII: e0301340

Researcher Affiliations

O'Kennedy, Martha M
  • Council for Scientific and Industrial Research (CSIR), Chemical Cluster, Pretoria, South Africa.
Roth, Robyn
  • Council for Scientific and Industrial Research (CSIR), Chemical Cluster, Pretoria, South Africa.
Ebersohn, Karen
  • Department of Veterinary Tropical Diseases, University of Pretoria, Pretoria, South Africa.
du Plessis, Lissinda H
  • Centre of Excellence for Pharmaceutical Sciences (PharmacenTM), North-West University, Potchefstroom, South Africa.
Mamputha, Sipho
  • Council for Scientific and Industrial Research (CSIR), Chemical Cluster, Pretoria, South Africa.
Rutkowska, Daria A
  • Council for Scientific and Industrial Research (CSIR), Chemical Cluster, Pretoria, South Africa.
du Preez, Ilse
  • Council for Scientific and Industrial Research (CSIR), Chemical Cluster, Pretoria, South Africa.
Verschoor, Jan A
  • Department of Biochemistry, University of Pretoria, Pretoria, South Africa.
Lemmer, Yolandy
  • Council for Scientific and Industrial Research (CSIR), Chemical Cluster, Pretoria, South Africa.

MeSH Terms

  • Animals
  • Mice
  • Horses
  • African Horse Sickness Virus / genetics
  • African Horse Sickness / prevention & control
  • Vaccines, Combined
  • Chromatography, Liquid
  • Capsid Proteins
  • Tandem Mass Spectrometry
  • Viral Vaccines
  • Antibodies, Viral

Conflict of Interest Statement

The authors have read the journal’s policy and have the following competing interests: Plant-produced chimaeric Orbivirus VLPs is patent protected (PCT/IB2017/052236, WO 2017/182958 A1, US 11,053,509 B2, ARIPO AP 6697 registered, invented by authors DR and MMO). The AHSV VP2 fusions proteins are protected by a patent application (PCT/IB2023/058808, invented by MMO and YL). This does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no additional patents, products in development or marketed products associated with this research to declare

References

This article includes 32 references
  1. Roy P. Orbivirus structure and assembly. Virology 1996;216:1–11.
    doi: 10.1006/viro.1996.0028pubmed: 8614976google scholar: lookup
  2. Rodríguez M, Joseph S, Pfeffer M, Raghavan R, Wernery U. Immune response of horses to inactivated African horse sickness vaccines. BMC Veterinary Research 2020;16:322.
    doi: 10.1186/s12917-020-02540-ypmc: PMC7466525pubmed: 32873300google scholar: lookup
  3. Calvo-Pinilla E, Gubbins S, Mertens PPC, Ortego J, Castillo-Olivares J. The immunogenicity of recombinant vaccines based on modified Vaccinia Ankara (MVA) viruses expressing African horse sickness virus VP2 antigens depends on the levels of expressed VP2 protein delivered to the host. Antiviral Research 2018;154:132–139.
    doi: 10.1016/j.antiviral.2018.04.015pmc: PMC5966619pubmed: 29678552google scholar: lookup
  4. Kanai YK, van Rijn PA, Maris-Veldhuis M, Kaname Y, Athmaram TN. Immunogenicity of recombinant VP2 proteins of all nine serotypes of African horse sickness virus. Vaccine 2014;32:4932–4937.
    doi: 10.1016/j.vaccine.2014.07.031pmc: PMC4148702pubmed: 25045805google scholar: lookup
  5. Stone-Marchat MA, Moss SR, Burrage TG, Barber ML, Roy P, Laegreid WW. Immunization with VP2 Is Sufficient for Protection against Lethal Challenge with African Horse sickness Virus Type 4. Virology 1996;220:219–222.
    doi: 10.1006/viro.1996.0304pubmed: 8659117google scholar: lookup
  6. Manning NM, Bachanek-Bankowska K, Mertens PPC, Castillo-Olivares J. Vaccination with recombinant Modified Vaccinia Ankara (MVA) viruses expressing single African horse sickness virus VP2 antigens induced cross-reactive virus neutralising antibodies (VNAb) in horses when administered in combination. Vaccine 2017;35:6024–6029.
    doi: 10.1016/j.vaccine.2017.04.005pubmed: 28438410google scholar: lookup
  7. Martínez-Torrecuadrada JL, Diaz-Laviada M, Roy P, Sanchez C, Vela C, Sanchez Vizcaino JM. Full protection against African horsesickness (AHS) in horses induced by baculovirus-derived AHS virus serotype 4 VP2, VP5 and VP7. J Gen Virol 1996;77:1211–1221.
    doi: 10.1099/0022-1317-77-6-1211pubmed: 8683209google scholar: lookup
  8. O’Kennedy MM, Coetzee P, Koekemoer O, du Plessis L, Lourens CW, Kwezi L. Protective immunity of plant-produced African horse sickness virus serotype 5 chimaeric virus-like particles (VLPs) and viral protein 2 (VP2) vaccines in IFNAR-/- mice. Vaccine 2022;40:5160–5169.
    doi: 10.1016/j.vaccine.2022.06.079pubmed: 35902279google scholar: lookup
  9. Crisci E, Bárcena J, Montoya M. Virus-like particle-based vaccines for animal viral infections. Immunología 2013;32(3):102–116.
    doi: 10.1016/j.inmuno.2012.08.002pmc: PMC7115488pubmed: 32287712google scholar: lookup
  10. Scanlen M, Paweska JT, Verschoor JA, van Dijk A. The protective efficacy of a recombinant VP2-based African horsesickness subunit vaccine candidate is determined by adjuvant. Vaccine 2002;20:1079–1088.
    doi: 10.1016/s0264-410x(01)00445-5pubmed: 11803068google scholar: lookup
  11. Dennis SJ, Meyers AE, Guthrie AJ, Hitzeroth II, Rybicki EP. Immunogenicity of plant-produced African horse sickness virus-like particles: implications for a novel vaccine. Plant Biotechnology Journal 2017;1–9.
    doi: 10.1111/pbi.12783pmc: PMC5787833pubmed: 28650085google scholar: lookup
  12. Dennis SJ, O’Kennedy MM, Rutkowska D, Tsekoa T, Lourens CW, Hitzeroth II. Safety and immunogenicity of plant-produced African horse sickness virus-like particles in horses. Veterinary Research 2019;49,105.
    doi: 10.1186/s13567-018-0600-4pmc: PMC6389048pubmed: 30309390google scholar: lookup
  13. Rutkowska DA, Mokoena N, Tsekoa TL, Dibakwane V, O’Kennedy MM. Plant produced chimaeric VLPs ‐ a new generation vaccine against African Horse Sickness. BMC Veterinary Research 2019;15:432.
    doi: 10.1186/s12917-019-2184-2pmc: PMC6892175pubmed: 31796116google scholar: lookup
  14. Martínez-Torrecuadrada J, Langeveld JPM, Meloen RH, Casal JI. Definition of neutralising sites on African horse sickness virus serotype 4 VP2 at the level of peptides. Journal of General Virology 2001;82:2415–2424.
    doi: 10.1099/0022-1317-82-10-2415pubmed: 11562535google scholar: lookup
  15. Aksular M, Calvo-Pinilla E, Marín-López A, Ortego J, Chambers AC, King LA, Castillo-Olivares J. A single dose of African horse sickness virus (AHSV) VP2 based vaccines provides complete clinical protection in a mouse model. Vaccine 2018;36:7003–7010.
    doi: 10.1016/j.vaccine.2018.09.065pmc: PMC6219453pubmed: 30309744google scholar: lookup
  16. Calvo-Pinilla E, de la Poza F, Gubbins S, Mertens PPC, Ortego J, Castillo-Olivares J. Vaccination of mice with a modified Vaccinia Ankara (MVA) virus expressing the African horse sickness virus (AHSV) capsid protein VP2 induces virus neutralising antibodies that confer protection against AHSV upon passive immunisation.. Virus Research 2014;180:23–30.
    doi: 10.1016/j.virusres.2013.12.002pubmed: 24333835google scholar: lookup
  17. Calvo-Pinilla E, de la Poza F, Gubbins S, Mertens PPC, Ortego J, Castillo-Olivares J. Antiserum from mice vaccinated with modified vaccinia Ankara virus expressing African horse sickness virus (AHSV) VP2 provides protection when it is administered 48 h before, or 48 h after challenge.. Antiviral Research 2015;116: 27–33.
    doi: 10.1016/j.antiviral.2015.01.009pmc: PMC7125940pubmed: 25643968google scholar: lookup
  18. Alberca B, Bachanek-Bankowska K, Cabana M, Calvo-Pinilla E, Viaplana E, Frost L. Vaccination of horses with a recombinant modified vaccinia Ankara virus (MVA) expressing African horse sickness (AHS) virus major capsid protein VP2 provides complete clinical protection against challenge.. Vaccine 2014; 32: 3670–3674.
    doi: 10.1016/j.vaccine.2014.04.036pmc: PMC4061461pubmed: 24837765google scholar: lookup
  19. De la Poza F, Marín-López A, Castillo-Olivares J, Calvo-Pinilla E, Ortego J. Identification of CD8 T cells epitopes in VP2 and NS1 proteins of African horse sickness virus in IFNAR (-/-) mice.. Virus Research 2015; 210: 149–153.
    doi: 10.1016/j.virusres.2015.08.005pubmed: 26272673google scholar: lookup
  20. Strasser R, Stadlmann J, Schähs M, Stiegler G, Quendler H, Mach L. Generation of glyco-engineered Nicotiana benthamiana for the production of monoclonal antibodies with a homogeneous human-like N-glycan structure.. Plant Biotechnology Journal 2008;6,392–402.
    doi: 10.1111/j.1467-7652.2008.00330.xpubmed: 18346095google scholar: lookup
  21. Von Teichman BF, Dungu B, Smit TK. cross-protection to African Horse Sickness serotypes 5 and 9 after vaccination with serotypes 8 and 6.. Vaccine 2010;28:6505–6517.
    doi: 10.1016/j.vaccine.2010.06.105pubmed: 20638456google scholar: lookup
  22. Weyer CT, Grewar JD, Burger P, Joone C, Lourens C, MacLachlan NJ. Dynamics of African horse sickness virus nucleic acid and antibody in horses following immunization with a commercial polyvalent live attenuated vaccine.. Vaccine 2017;35:2504–2510.
    doi: 10.1016/j.vaccine.2017.03.005pubmed: 28341113google scholar: lookup
  23. Chiam R, Sharp E, Maan S, Rao S, Mertens PPC, Blacklaws B. Induction of antibody responses to African Horse sickness virus (AHSV) in ponies after vaccination with recombinant modified vaccinia Ankara (MVA).. PLoS ONE 2009; 4(6):5997.
    doi: 10.1371/journal.pone.0005997pmc: PMC2694985pubmed: 19543394google scholar: lookup
  24. Guthrie AJ, Quan M, Lourens CW, Audonnet J-C, Minke JM, Yao J. Protective immunization of horses with a recombinant canarypox virus vectored vaccine co-expressing gene encoding the outer capsid proteins of African horse sickness virus.. Vaccine 2009;27:4434–4438.
    doi: 10.1016/j.vaccine.2009.05.044pubmed: 19490959google scholar: lookup
  25. Roy P, Bishop DHL, Howard S, Aitchison H, Erasmus B. Recombinant baculovirus-synthesised African horsesickness virus (AHSV) outer-capsid protein VP2 provides protection against lethal AHSV challenge.. J Gen Virol 1996;77:2053–7.
    doi: 10.1099/0022-1317-77-9-2053pubmed: 8811002google scholar: lookup
  26. Castillo-Olivares J, Calvo-Pinilla E, Casanova I, Bachanek-Bankowska K, Chiam R, Maan S. A modified Vaccinia Ankara Virus (MVA) vaccine expressing African horse sickness virus (AHSV) VP2 protects against AHSV challenge in an IFNAR-/- mouse model.. PlosOne 2011;6(1):16503.
    doi: 10.1371/journal.pone.0016503pmc: PMC3027694pubmed: 21298069google scholar: lookup
  27. Lua LHL, Connors NK, Sainsbury F, Chuan YP, Wibowo N, Middelberg APJ. Bioengineering Virus-Like Particles as Vaccines.. 2013;111,3: 425–440.
    doi: 10.1002/bit.25159pubmed: 24347238google scholar: lookup
  28. Crafford JE, Lourens CW, Smit TK, Gardner IA, MacLachlan NJ, Guthrie AJ. Serological response of foals to polyvalent and monovalent live-attenuated African horse sickness virus vaccines.. Vaccine 2014;32:3611–3616.
    doi: 10.1016/j.vaccine.2014.04.087pubmed: 24814557google scholar: lookup
  29. Van Rijn PA, Maris-Veldmuis M, Grobler M, Wright IM, Erasmus BJ, Maartens LH. Safety and efficacy of inactivated African horse sickness (AHS) vaccine formulated with different adjuvants.. Vaccine 2020;38: 7108–7117.
    doi: 10.1016/j.vaccine.2020.08.072pubmed: 32921506google scholar: lookup
  30. Huber VC, McKeon RM, Brackin MN, Miller LA, Keating R, Brown SA. Distinct Contributions of Vaccine-Induced Immunoglobulin G1 (IgG1) and IgG2a Antibodies to Protective Immunity against Influenza.. Clinical and Vaccine Immunology 2006;13(9):981–990.
    doi: 10.1128/CVI.00156-06pmc: PMC1563571pubmed: 16960108google scholar: lookup
  31. van Rijn PA. Prospects of next-generation vaccines for Bluetongue.. Frontiers in Veterinary Science 2019;6:407.
    doi: 10.3389/fvets.2019.00407pmc: PMC6881303pubmed: 31824966google scholar: lookup
  32. O’Kennedy MM, Abolnik C, Smith T, Motlou T, Goosen K, Sepotokele KM. Immunogenicity of adjuvanted plant-produced SARS-CoV-2 Beta spike VLP vaccine in New Zealand white rabbits.. Vaccine 2022;41:2261–2269.
    doi: 10.1016/j.vaccine.2023.02.050pmc: PMC9968623pubmed: 36868876google scholar: lookup

Citations

This article has been cited 2 times.
  1. Tinarwo M, Dennis SJ, Hitzeroth II, Meyers AE, Rybicki EP, Mbewana S. Development of an African horse sickness VP6 DIVA diagnostic ELISA.. Virol J 2025 Aug 12;22(1):276.
    doi: 10.1186/s12985-025-02898-1pubmed: 40796889google scholar: lookup
  2. Pitchers KG, Boakye OD, Campeotto I, Daly JM. The Potential of Plant-Produced Virus-like Particle Vaccines for African Horse Sickness and Other Equine Orbiviruses.. Pathogens 2024 May 28;13(6).
    doi: 10.3390/pathogens13060458pubmed: 38921755google scholar: lookup
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