Influenza virus‐like particles produced by transient expression in Nicotiana benthamiana induce a protective immune response against a lethal viral challenge in mice
Open Access
- 12 November 2008
- journal article
- research article
- Published by Wiley in Plant Biotechnology Journal
- Vol. 6 (9), 930-940
- https://doi.org/10.1111/j.1467-7652.2008.00384.x
Abstract
A strain‐specific vaccine represents the best possible response to the threat of an influenza pandemic. Rapid delivery of such a vaccine to the world's population before the peak of the first infection wave seems to be an unattainable goal with the current influenza vaccine manufacturing capacity. Plant‐based transient expression is one of the few production systems that can meet the anticipated surge requirement. To assess the capability of plant agroinfiltration to produce an influenza vaccine, we expressed haemagglutinin (HA) from strains A/Indonesia/5/05 (H5N1) and A/New Caledonia/20/99 (H1N1) by agroinfiltration of Nicotiana benthamiana plants. Size distribution analysis of protein content in infiltrated leaves revealed that HA was predominantly assembled into high‐molecular‐weight structures. H5‐containing structures were purified and examination by transmission electron microscopy confirmed virus‐like particle (VLP) assembly. High‐performance thin layer chromatography analysis of VLP lipid composition highlighted polar and neutral lipid contents comparable with those of purified plasma membranes from tobacco plants. Electron microscopy of VLP‐producing cells in N. benthamiana leaves confirmed that VLPs accumulated in apoplastic indentations of the plasma membrane. Finally, immunization of mice with two doses of as little as 0.1 µg of purified influenza H5‐VLPs triggered a strong immune response against the homologous virus, whereas two doses of 0.5 µg of H5‐VLPs conferred complete protection against a lethal challenge with the heterologous A/Vietnam/1194/04 (H5N1) strain. These results show, for the first time, that plants are capable of producing enveloped influenza VLPs budding from the plasma membrane; such VLPs represent very promising candidates for vaccination against influenza pandemic strains.Keywords
This publication has 27 references indexed in Scilit:
- Cross-Clade Protective Immune Responses to Influenza Viruses with H5N1 HA and NA Elicited by an Influenza Virus-Like ParticlePLOS ONE, 2008
- A plant‐produced influenza subunit vaccine protects ferrets against virus challengeInfluenza and Other Respiratory Viruses, 2008
- Influenza Virus Hemagglutinin and Neuraminidase, but Not the Matrix Protein, Are Required for Assembly and Budding of Plasmid-Derived Virus-Like ParticlesJournal of Virology, 2007
- Virus-Like Particle Vaccine Induces Protective Immunity against Homologous and Heterologous Strains of Influenza VirusJournal of Virology, 2007
- Characterization of Lipid Rafts from Medicago truncatula Root Plasma Membranes: A Proteomic Study Reveals the Presence of a Raft-Associated Redox SystemPlant Physiology, 2007
- Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectorsProceedings of the National Academy of Sciences of the United States of America, 2006
- Virus-like particles: Passport to immune recognitionMethods, 2006
- Virus-like particles production in green plantsMethods, 2006
- Virus-Like Particle (VLP) Vaccine Conferred Complete Protection against a Lethal Influenza Virus ChallengeViral Immunology, 2005
- Assembly and budding of influenza virusVirus Research, 2004