title: High immunogenicity of plant-produced candidate influenza vaccine based on the M2e peptide fused to flagellin.
abstract: The ectodomain of the conserved influenza matrix protein M2 (M2e) is a promising target for the development of a universal influenza vaccines. Immunogenicity of M2e could be enhanced by its fusion to bacterial flagellin, the ligand for Toll-like receptor 5. Previously we reported the transient expression in plants of a recombinant protein Flg-4M comprising flagellin fused to 4 tandem copies of the M2e. The use of self-replicating recombinant vector based on the potato virus X allowed expression of Flg-4M in Nicotiana benthaminana leaves at a very high level, up to about 1 mg/g of fresh leaf tissue. Intranasal immunization of mice with Flg-4M induced M2e-specific serum antibodies and provided protection against lethal challenge with different strains of influenza A virus. Here we show that immunization with Flg-4M not only generates a strong immune response, but also redirects the response from the carrier flagellin toward the M2e epitopes. Significant IgG response to M2e was also developed in bronchoalveolar lavages of immunized mice. Protective activity of Flg-4M upon lethal influenza challenge correlated with a decrease of virus titers in lungs relative to the control. Overall these data show the potential for the development of a plant-produced M2e-flagellin universal influenza vaccine.
authors.affiliation: a Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences , Moscow , Russia.|b Research Institute of Influenza, Russian Federation Ministry of Health , St. Petersburg , Russia.
authors.forename: Eugenia S
authors.initials: ES
authors.lastname: Mardanova
authors.affiliation: a Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences , Moscow , Russia.|b Research Institute of Influenza, Russian Federation Ministry of Health , St. Petersburg , Russia.
authors.forename: Roman Y
authors.initials: RY
authors.lastname: Kotlyarov
authors.affiliation: a Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences , Moscow , Russia.
authors.forename: Victor V
authors.initials: VV
authors.lastname: Kuprianov
authors.affiliation: b Research Institute of Influenza, Russian Federation Ministry of Health , St. Petersburg , Russia.
authors.forename: Liudmila A
authors.initials: LA
authors.lastname: Stepanova
authors.affiliation: b Research Institute of Influenza, Russian Federation Ministry of Health , St. Petersburg , Russia.
authors.forename: Liudmila M
authors.initials: LM
authors.lastname: Tsybalova
authors.affiliation: c Department of Biological Chemistry , John Innes Center, Norwich Research Park , Norwich , UK.
authors.forename: George P
authors.initials: GP
authors.lastname: Lomonossoff
authors.affiliation: a Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences , Moscow , Russia.
authors.forename: Nikolai V
authors.initials: NV
authors.lastname: Ravin
mesh_terms: D000281:Administration, Intranasal; D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D000091344:Filaggrin Proteins; D005408:Flagellin / Q000235:genetics / Q000276:immunology*; D005822:Genetic Vectors / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D000071497:Immunogenicity, Vaccine; D007074:Immunoglobulin G / Q000096:biosynthesis; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000276:immunology*; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000096:biosynthesis* / Q000235:genetics; D007251:Influenza, Human / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D017863:Potexvirus / Q000235:genetics; D000072417:Protein Domains; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism; D014611:Vaccination; D014763:Viral Matrix Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007074:Immunoglobulin G; D007252:Influenza Vaccines; C488936:M1 protein, Influenza A virus; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; D005408:Flagellin
keywords: M2e peptide; flagellin; influenza; plant-produced vaccine