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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

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Citations

This article has been cited 2 times.
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  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).
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