title: Towards a novel influenza vaccine: engineering of hemagglutinin on a platform of adenovirus dodecahedron.
abstract: BACKGROUND
  The production process for the current influenza vaccine takes about 6 months and its antigenic composition must be modified annually. In the attempt towards developing influenza vaccine production that would be faster, safer and cheaper we engineered an influenza vaccine in which multiple copies of hemagglutinin (HA) would be delivered by a vector, adenovirus dodecahedron (Ad Dd). Dd is a virus-like particle, formed by assembly of twelve copies of pentameric penton base (Pb) proteins responsible for virus penetration. In order to attach HA to the vector, an adaptor containing WW domains was used. The WW domain is a linear peptide fragment identified as a partner of proline-proline-x-tyrosine (PPxY) motif present at the N-terminal extremity of the Pb protein, which is a building block of Dd. That tandem of three WW domains in fusion with the protein of interest enables interaction with Dd and efficient translocation to the cytoplasm of cells in culture.
  
  
  RESULTS
  Since HA is an oligomeric protein with complicated processing, we prepared six different constructs of HA (A/swan/Poland/467/2006(H5N1)) in fusion with the WW adaptor. Herein we report baculovirus expression and functional analysis of six HA-WW variants. The best behaving variant was successfully delivered into human cells in vitro.
  
  
  CONCLUSIONS
  Engineering of a soluble complex of HA with Dd, a virus-like particle that serves as a vector, an adjuvant and as a multivalent presentation platform, is an important step toward a novel influenza vaccine.
authors.affiliation: Institute of Biochemistry and Biophysics, PAS, Pawinskiego 5a, 02106 Warsaw, Poland.
authors.forename: Antonina
authors.initials: A
authors.lastname: Naskalska
authors.forename: Ewa
authors.initials: E
authors.lastname: Szolajska
authors.forename: Igor
authors.initials: I
authors.lastname: Andreev
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Podsiadla
authors.forename: Jadwiga
authors.initials: J
authors.lastname: Chroboczek
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics; D006383:Hemadsorption Inhibition Tests; D006388:Hemagglutinins / Q000235:genetics* / Q000378:metabolism; D007252:Influenza Vaccines / Q000235:genetics* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D043743:Ubiquitin-Protein Ligase Complexes / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D056827:Endosomal Sorting Complexes Required for Transport; D006388:Hemagglutinins; D007252:Influenza Vaccines; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014764:Viral Proteins; D043743:Ubiquitin-Protein Ligase Complexes; C469287:RSP5 protein, S cerevisiae