An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particles.
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.
Santi
Luca
L
The Biodesign Institute and School of Life Sciences, Arizona State University, Tempe, AZ 85287-5401, USA.
Batchelor
Lance
L
Huang
Zhong
Z
Hjelm
Brooke
B
Kilbourne
Jacquelyn
J
Arntzen
Charles J
CJ
Chen
Qiang
Q
Mason
Hugh S
HS
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
D007070:Immunoglobulin A; D022462:Vaccines, Edible; D022721:Vaccines, Virosome; D014765:Viral Vaccines; D022701:Virosomes