title: Protection against multiple influenza A virus strains induced by candidate recombinant vaccine based on heterologous M2e peptides linked to flagellin.
abstract: Matrix 2 protein ectodomain (M2e) is considered a promising candidate for a broadly protective influenza vaccine. M2e-based vaccines against human influenza A provide only partial protection against avian influenza viruses because of differences in the M2e sequences. In this work, we evaluated the possibility of obtaining equal protection and immune response by using recombinant protein on the basis of flagellin as a carrier of the M2e peptides of human and avian influenza A viruses. Recombinant protein was generated by the fusion of two tandem copies of consensus M2e sequence from human influenza A and two copies of M2e from avian A/H5N1 viruses to flagellin (Flg-2M2eh2M2ek). Intranasal immunisation of Balb/c mice with recombinant protein significantly elicited anti-M2e IgG in serum, IgG and sIgA in BAL. Antibodies induced by the fusion protein Flg-2M2eh2M2ek bound efficiently to synthetic peptides corresponding to the human consensus M2e sequence as well as to the M2e sequence of A/Chicken/Kurgan/05/05 RG (H5N1) and recognised native M2e epitopes exposed on the surface of the MDCK cells infected with A/PR/8/34 (H1N1) and A/Chicken/Kurgan/05/05 RG (H5N1) to an equal degree. Immunisation led to both anti-M2e IgG1 and IgG2a response with IgG1 prevalence. We observed a significant intracellular production of IL-4, but not IFN-γ, by CD4+ T-cells in spleen of mice following immunisation with Flg-2M2eh2M2ek. Immunisation with the Flg-2M2eh2M2ek fusion protein provided similar protection from lethal challenge with human influenza A viruses (H1N1, H3N2) and avian influenza virus (H5N1). Immunised mice experienced significantly less weight loss and decreased lung viral titres compared to control mice. The data obtained show the potential for the development of an M2e-flagellin candidate influenza vaccine with broad spectrum protection against influenza A viruses of various origins.
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Liudmila A
authors: LA
authors: Stepanova
authors: Centre "Bioengineering", Russian Academy of Sciences, Moscow, Russia.
authors: Roman Y
authors: RY
authors: Kotlyarov
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Anna A
authors: AA
authors: Kovaleva
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Marina V
authors: MV
authors: Potapchuk
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Alexandr V
authors: AV
authors: Korotkov
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Mariia V
authors: MV
authors: Sergeeva
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Marina A
authors: MA
authors: Kasianenko
authors: Centre "Bioengineering", Russian Academy of Sciences, Moscow, Russia.
authors: Victor V
authors: VV
authors: Kuprianov
authors: Centre "Bioengineering", Russian Academy of Sciences, Moscow, Russia; GenNanotech Ltd, St. Petersburg, Russia.
authors: Nikolai V
authors: NV
authors: Ravin
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Liudmila M
authors: LM
authors: Tsybalova
authors: Centre "Bioengineering", Russian Academy of Sciences, Moscow, Russia.
authors: Konstantin G
authors: KG
authors: Skryabin
authors: Department of Influenza Vaccines, Research Institute of Influenza, Ministry of Health of the Russian Federation, St. Petersburg, Russia.
authors: Oleg I
authors: OI
authors: Kiselev
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D000091344:Filaggrin Proteins; D005408:Flagellin / Q000276:immunology*; D006801:Humans; D007114:Immunization; D009980:Influenza A virus / Q000276:immunology*; D007252:Influenza Vaccines / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D008168:Lung / Q000276:immunology / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D019562:Viral Load; D014763:Viral Matrix Proteins / Q000276:immunology*; D000086082:Viroporin Proteins
chemical_list: D000914:Antibodies, Viral; D000939:Epitopes; D000091344:Filaggrin Proteins; D005408:Flagellin; D007252:Influenza Vaccines; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014763:Viral Matrix Proteins; C000717249:FLG protein, human; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins