title: An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles.
abstract: Virus-like particles (VLPs) derived from enteric pathogens like Norwalk virus (NV) are well suited to study oral immunization. We previously described stable transgenic plants that accumulate recombinant NV-like particles (rNVs) that were orally immunogenic in mice and humans. The transgenic approach suffers from long generation time and modest level of antigen accumulation. We now overcome these constraints with an efficient tobacco mosaic virus (TMV)-derived transient expression system using leaves of Nicotiana benthamiana. We produced properly assembled rNV at 0.8 mg/g leaf 12 days post-infection (dpi). Oral immunization of CD1 mice with 100 or 250 microg/dose of partially purified rNV elicited systemic and mucosal immune responses. We conclude that the plant viral transient expression system provides a robust research tool to generate abundant quantities of rNV as enriched, concentrated VLP preparations that are orally immunogenic.
authors.affiliation: The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85287-5401, USA.
authors.forename: Luca
authors.initials: L
authors.lastname: Santi
authors.forename: Lance
authors.initials: L
authors.lastname: Batchelor
authors.forename: Zhong
authors.initials: Z
authors.lastname: Huang
authors.forename: Brooke
authors.initials: B
authors.lastname: Hjelm
authors.forename: Jacquelyn
authors.initials: J
authors.lastname: Kilbourne
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Arntzen
authors.forename: Qiang
authors.initials: Q
authors.lastname: Chen
authors.forename: Hugh S
authors.initials: HS
authors.lastname: Mason
mesh_terms: D000818:Animals; D017250:Caliciviridae Infections / Q000517:prevention & control*; D005260:Female; D005759:Gastroenteritis / Q000517:prevention & control*; D007070:Immunoglobulin A / Q000032:analysis; D007422:Intestines / Q000276:immunology; D051379:Mice; D046529:Microscopy, Electron, Transmission; D009663:Norwalk virus / Q000235:genetics / Q000276:immunology*; D014026:Nicotiana / Q000235:genetics / Q000276:immunology*; D014027:Tobacco Mosaic Virus / Q000235:genetics*; D022462:Vaccines, Edible / Q000235:genetics / Q000276:immunology; D022721:Vaccines, Virosome / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology*; D022701:Virosomes / Q000648:ultrastructure
chemical_list: D007070:Immunoglobulin A; D022462:Vaccines, Edible; D022721:Vaccines, Virosome; D014765:Viral Vaccines; D022701:Virosomes