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.
Highlights
Noroviruses (NoVs), which belong to the family Caliciviridae, are the leading causes of acute viral gastroenteritis in humans worldwide
The Narita virus capsid protein (NaVCP) coding sequence was cloned into the 3 module, and when delivered in combination with 5 module and integrase module, the system drives the expression of NaVCP in the cytoplasm
Three days postinfiltration (DPI), total RNA was extracted from the N. benthamiana leaf and RNA blot was performed with 3UTR specific probe
Summary
Noroviruses (NoVs), which belong to the family Caliciviridae, are the leading causes of acute viral gastroenteritis in humans worldwide. The open reading frame (ORF) 1 encodes nonstructural proteins required for viral replication. The major capsid protein can diverge by as much as 60% between genogroups and 20– 30% between genotypes within a genogroup. The majority of norovirus outbreaks currently are caused by the GII. genotypes, which cause significant morbidity and mortality [6]. Several viruses in this category have emerged since 1990s worldwide [7,8,9,10,11,12,13,14,15]. We studied the development of a plant-based vaccine against Narita 104 virus (NaV) (NCBI database, accession number AB078336), a GII. strain of norovirus [16]
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