Boosting in Planta Production of Antigens Derived From the Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Subsequent Evaluation of Their Immunogenicity

PLoS One. 2014 Mar 10;9(3):e91386. doi: 10.1371/journal.pone.0091386. eCollection 2014.

Abstract

Porcine reproductive and respiratory syndrome (PRRS) is a disease of swine, caused by an arterivirus, the PRRS virus (PRRSV). This virus infects pigs worldwide and causes huge economic losses. Due to genetic drift, current vaccines are losing their power. Adaptable vaccines could provide a solution to this problem. This study aims at producing in planta a set of antigens derived from the PRRSV glycoproteins (GPs) to be included in a subunit vaccine. We selected the GP3, GP4 and GP5 and optimized these for production in an Arabidopsis seed platform by removing transmembrane domains (Tm) and/or adding stabilizing protein domains, such as the green fluorescent protein (GFP) and immunoglobulin (IgG) 'Fragment crystallizable' (Fc) chains. Accumulation of the GPs with and without Tm was low, reaching no more than 0.10% of total soluble protein (TSP) in homozygous seed. However, addition of stabilizing domains boosted accumulation up to a maximum of 2.74% of TSP when GFP was used, and albeit less effectively, also the Fc chains of the porcine IgG3 and murine IgG2a increased antigen accumulation, to 0.96% and 1.81% of TSP respectively, while the murine IgG3 Fc chain did not. Antigens with Tm were less susceptible to these manipulations to increase yield. All antigens were produced in the endoplasmic reticulum and accordingly, they carried high-mannose N-glycans. The immunogenicity of several of those antigens was assessed and we show that vaccination with purified antigens did elicit the production of antibodies with virus neutralizing activity in mice but not in pigs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Viral / biosynthesis
  • Antigens, Viral / immunology*
  • Antigens, Viral / isolation & purification
  • Arabidopsis / genetics
  • Arabidopsis / metabolism*
  • Chromatography, Affinity
  • DNA, Bacterial / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Glycoproteins / immunology*
  • Glycosylation
  • Mice, Inbred C57BL
  • Models, Animal
  • Molecular Sequence Data
  • Plants, Genetically Modified
  • Polysaccharides / metabolism
  • Porcine respiratory and reproductive syndrome virus / immunology*
  • Seeds / metabolism
  • Swine / virology*

Substances

  • Antigens, Viral
  • DNA, Bacterial
  • Glycoproteins
  • Polysaccharides
  • T-DNA

Associated data

  • GENBANK/KF983786
  • GENBANK/KF983787
  • GENBANK/KF983788
  • GENBANK/KF983789
  • GENBANK/KF983790
  • GENBANK/KF983791
  • GENBANK/KF983792
  • GENBANK/KF983793
  • GENBANK/KF983794
  • GENBANK/KF983795
  • GENBANK/KF983796
  • GENBANK/KF983797
  • GENBANK/KF983798
  • GENBANK/KF983799
  • GENBANK/KF983800
  • GENBANK/KF983801
  • GENBANK/KF983802
  • GENBANK/KF983803
  • GENBANK/KF983804

Grant support

Robin Piron received a predoctoral scholarship for Strategic Basic Research from the Agency for Innovation by Science and Technology (IWT). Stefaan De Koker received a postdoctoral grant from the Fonds voor Wetenschappelijk Onderzoek (FWO). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.