Immunogenicity of plant-produced African horse sickness virus-like particles : implications for a novel vaccine

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Authors

Dennis, Susan J.
Meyers, Ann E.
Guthrie, Alan John
Hitzeroth, Inga I.
Rybicki, Edward P.

Journal Title

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Volume Title

Publisher

Wiley

Abstract

African horse sickness (AHS) is a debilitating and often fatal viral disease affecting horses in much of Africa, caused by the dsRNA orbivirus African horse sickness virus (AHSV). Vaccination remains the single most effective weapon in combatting AHS, as there is no treatment for the disease apart from good animal husbandry. However, the only commercially available vaccine is a liveattenuated version of the virus (LAV). The threat of outbreaks of the disease outside its endemic region and the fact that the LAV is not licensed for use elsewhere in the world, have spurred attempts to develop an alternative safer, yet cost-effective recombinant vaccine. Here, we report the plant-based production of a virus-like particle (VLP) AHSV serotype five candidate vaccine by Agrobacterium tumefaciens-mediated transient expression of all four capsid proteins in Nicotiana benthamiana using the cowpea mosaic virus-based HyperTrans (CPMV-HT) and associated pEAQ plant expression vector system. The production process is fast and simple, scalable, economically viable, and most importantly, guinea pig antiserum raised against the vaccine was shown to neutralize live virus in cell-based assays. To our knowledge, this is the first report of AHSV VLPs produced in plants, which has important implications for the containment of, and fight against the spread of, this deadly disease.

Description

Figure S1 Optimization of plant-based expression of recombinant AHSV-5 structural proteins.
Figure S2 Increased formation of AHSV-5 VLPs incorporating a mutated version of VP7.
Figure S3 Purification of AHSV-5 VLPs.
Figure S4 Mass spectrometry analysis of the 4 protein bands recovered from SDS-PAGE separation of density gradient fractions from leaves co-infiltrated with Agrobacterium AGL1 pEAQ recombinants for co-expression of AHSV capsid proteins VP2, VP3, VP5 and VP7mu.
Figure S5 Quantification of AHSV-5 VLPs by gel densitometry.
Table S1 Virus neutralizing antibody titres of serum samples from vaccinated and control guinea pigs V1 and C1.

Keywords

Biopharming, Transient expression, Vaccines, African horse sickness (AHS), African horse sickness virus (AHSV), Virus-like particle (VLP), Cowpea mosaic virus-based HyperTrans (CPMV-HT)

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Citation

Dennis, S. J., Meyers, A. E., Guthrie, A. J., Hitzeroth, I. I. and Rybicki, E. P. (2018), Immunogenicity of plant-produced African horse sickness virus-like particles: implications for a novel vaccine. Plant Biotechnol J, 16: 442–450. doi:10.1111/pbi.12783.