Norovirus Narita 104 Virus-Like Particles Expressed inNicotiana benthamianaInduce Serum and Mucosal Immune Responses

Abstract

Narita 104 virus is a human pathogen belonging to the norovirus (family Caliciviridae) genogroup II. Noroviruses cause epidemic gastroenteritis worldwide. To explore the potential of developing a plant-based vaccine, a plant optimized gene encoding Narita 104 virus capsid protein (NaVCP) was expressed transiently in Nicotiana benthamiana using a tobacco mosaic virus expression system. NaVCP accumulated up to approximately 0.3 mg/g fresh weight of leaf at 4 days postinfection. Initiation of hypersensitive response-like symptoms followed by tissue necrosis necessitated a brief infection time and was a significant factor limiting expression. Transmission electron microscopy of plant-derived NaVCP confirmed the presence of fully assembled virus-like particles (VLPs). In this study, an optimized method to express and partially purify NaVCP is described. Further, partially purified NaVCP was used to immunize mice by intranasal delivery and generated significant mucosal and serum antibody responses. Thus, plant-derived Narita 104 VLPs have potential for use as a candidate subunit vaccine or as a component of a multivalent subunit vaccine, along with other genotype-specific plant-derived VLPs.

Prospects and Challenges in the Development of a Norovirus Vaccine

“…Norovirus VLPs have been successfully generated in embryonated chicken eggs, mammalian cells, plant cells, and yeasts in various systems [63–68]. In many of these reports, the VLPs produced were used in animal vaccination studies and shown to be immunogenic as has been previously shown with traditionally produced VLPs [63, 65, 69]. At least one of these constructs has been tested in human subjects.…”

From Pandemic Preparedness to Biofuel Production: Tobacco Finds Its Biotechnology Niche in North America

“…Lastly, the DNA plasmid constructs engineered by researchers to be transferred from bacteria into plants have historically incorporated or built upon genetic elements of tobacco mosaic virus (TMV) or other plant viruses that replicate in tobacco leaves, therefore further justifying the use of tobacco as a plant host for expression of these target biotherapeutics. [Mallajosyula, 2014: 24378714], [Kalthoff, 2010: 20810729], [Kanagarajan, 2012: 22442675] Ebola virus [12][13][14][15] [ Phoolcharoen, 2011: 21281425], [Phoolcharoen, 2011: 22143779], [Huang, 2010, 20047189], [Castilho, 2011: 22039433] Dengue [16][17][18][19][20] [ Kim, 2015: 25728317], [Conconi-Linares, 2013: 23499580], [Saejung, 2007: 17659815], [Martinez, 2010: 20213522], [Martinez, 2012: 22480936] Norovirus [21][22][23][24][25] [ Mathew, 2014: 24949472], [Santi, 2008: 18325641], [Lai, 2012: 22134876 [Farrance, 2011: 21715576], [Feller, 2013: 24278216], [Kapelski, 2015: 25651860], [Voepel, 2014: 25200253], [Clemente, 2012: 22911156], [Beiss, 2015: 25615702], West Nile Virus [49][50][51] [ Chen, 2015: 25676782], [Lai, 2014: 24975464], 24675995] Bacillus anthracis (anthrax) [52][53][54][55][...…”

Meeting Report VLPNPV: Session 3: Immune responses

“…First, M. M Herbst-Kralovetz from the University of Arizona School of Medicine, USA, elaborated on her ongoing studies of norovirus VLPs as a vaccine. 12 Since the introduction of the rotavirus vaccine, norovirus, a positive stranded, nonenveloped virus classified in the Caliciviridae family, has become the predominant cause of acute viral diarrhea. The goal of the studies reported at this meeting was to optimize mucosal responses to a norovirus VLP produced in plants using a TMV vector.…”