title: A plant-produced H1N1 trimeric hemagglutinin protects mice from a lethal influenza virus challenge.
abstract: The increased worldwide awareness of seasonal and pandemic influenza, including pandemic H1N1 virus, has stimulated interest in the development of economic platforms for rapid, large-scale production of safe and effective subunit vaccines. In recent years, plants have demonstrated their utility as such a platform and have been used to produce vaccine antigens against various infectious diseases. Previously, we have produced in our transient plant expression system a recombinant monomeric hemagglutinin (HA) protein (HAC1) derived from A/California/04/09 (H1N1) strain of influenza virus and demonstrated its immunogenicity and safety in animal models and human volunteers. In the current study, to mimic the authentic HA structure presented on the virus surface and to improve stability and immunogenicity of the HA antigen, we generated trimeric HA by introducing a trimerization motif from a heterologous protein into the HA sequence. Here, we describe the engineering, production in Nicotiana benthamiana plants, and characterization of the highly purified recombinant trimeric HA protein (tHA-BC) from A/California/04/09 (H1N1) strain of influenza virus. The results demonstrate the induction of serum hemagglutination inhibition antibodies by tHA-BC and its protective efficacy in mice against a lethal viral challenge. In addition, the immunogenic and protective doses of tHA-BC were much lower compared with monomeric HAC1. Further investigation into the optimum vaccine dose and/or regimen as well as the stability of trimerized HA is necessary to determine whether trimeric HA is a more potent vaccine antigen than monomeric HA.
authors.affiliation: Fraunhofer USA Center for Molecular Biotechnology; Newark, DE USA.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Shoji
authors.forename: R Mark
authors.initials: RM
authors.lastname: Jones
authors.forename: Vadim
authors.initials: V
authors.lastname: Mett
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Chichester
authors.forename: Konstantin
authors.initials: K
authors.lastname: Musiychuk
authors.forename: Xiangjie
authors.initials: X
authors.lastname: Sun
authors.forename: Terrence M
authors.initials: TM
authors.lastname: Tumpey
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Green
authors.forename: Moneim
authors.initials: M
authors.lastname: Shamloul
authors.forename: Joey
authors.initials: J
authors.lastname: Norikane
authors.forename: Hong
authors.initials: H
authors.lastname: Bi
authors.forename: Caitlin E
authors.initials: CE
authors.lastname: Hartman
authors.forename: Cory
authors.initials: C
authors.lastname: Bottone
authors.forename: Michelle
authors.initials: M
authors.lastname: Stewart
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Streatfield
authors.forename: Vidadi
authors.initials: V
authors.lastname: Yusibov
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D004195:Disease Models, Animal; D006385:Hemagglutination Inhibition Tests; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology*; D008807:Mice, Inbred BALB C; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000517:prevention & control*; D030821:Plants, Genetically Modified / Q000235:genetics; D015202:Protein Engineering; D055503:Protein Multimerization; D016019:Survival Analysis; D014026:Nicotiana / Q000235:genetics; D016896:Treatment Outcome; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000276:immunology; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic
keywords: hemagglutinin; influenza; plant-produced antigen; recombinant protein; subunit vaccine; trimer