Development of a cucumber green mottle mosaic virus-based expression vector for the production in cucumber of neutralizing epitopes against a devastating animal virus

Abstract

Virus-based expression systems have been widely exploited for the production of recombinant proteins in plants during the last thirty years. Advances in technology have boosted scale-up manufacturing of plant-made pharmaceuticals to high levels, via the complementation of transient expression and viral vectors. This combination allows proteins of interest to be produced in plants within a matter of days and thus, is well suited for the development of plant-made vaccines or therapeutics against emerging infectious diseases and potential bioterrorism agents. Several plant-based products are currently in varying stages of clinical development. To investigate the viability of virus-based expression systems for plant-made vaccines against porcine reproductive and respiratory syndrome virus (PRRSV), the most devastating threat to the pork industry in Canada, we cloned the full-length genome of a cucumber green mottle mosaic virus (CGMMV) isolate and developed a CGMMV-based expression vector. We further employed this vector to express the neutralizing epitope (NE) of PRRSV glycoprotein 5 (GP5) in cucumber leaves via agroinfiltration. The coding region of the GP5 NE was inserted downstream of the open reading frame for coat protein (CP) and expressed by a readthrough mechanism. The chimeric virus particles were stable and the expression levels reached as high as 35.84 mg/kg of cucumber leaf fresh weight. This study offers a promising solution to the production of a low cost, versatile and robust vaccine for oral administration against PRRSV through a chimeric virus particle display system.

Keywords: CGMMV; Molecular farming; PRRSV; Particle display; Virus vector.