title: A fragment of the envelope protein from dengue-1 virus, fused in two different sites of the meningococcal P64k protein carrier, induces a functional immune response in mice.
abstract: Previously we have reported the capacity of the fusion protein PD3, composed of the P64k protein and the envelope (E) fragment from amino acids (aa) 286-426 of dengue-2 virus (DEN-2), to induce a functional immune response in mice against the homologous virus. In that case, the E fragment was inserted within the lipoyl-binding domain of the meningococcal P64k protein. In the present study, to test the functionality of the same E region from dengue-1 (DEN-1), a similar construct was made. Furthermore, another alternative of fusion protein was also constructed where the same E fragment from DEN-1 was fused to the C-terminus of the P64k protein. The recombinant proteins obtained (PD11 and PD10) were semi-purified and analysed for their antigenicity, immunogenicity and the ability to protect mice against lethal challenge. Both molecules exhibited the same recognition patterns against anti-DEN-1 polyclonal antibodies. In addition, when administered to mice, they elicited high levels of neutralizing antibodies and induced significant protection against lethal challenge with DEN-1 after intracerebral inoculation. These results reveal the availability of two sites within the P64k for the further insertion of DEN fragments, enabling a construct carrying two fragments from heterologous serotypes within the same molecule of this protein carrier.
authors.affiliation: Centro de Ingeniería Genética y Biotecnología, Habana, Cuba. lisset.hermida@cigb.edu.cu
authors.forename: Lisset
authors.initials: L
authors.lastname: Hermida
authors.forename: Rayner
authors.initials: R
authors.lastname: Rodríguez
authors.forename: Laura
authors.initials: L
authors.lastname: Lazo
authors.forename: Lídice
authors.initials: L
authors.lastname: Bernardo
authors.forename: Ricardo
authors.initials: R
authors.lastname: Silva
authors.forename: Aída
authors.initials: A
authors.lastname: Zulueta
authors.forename: Carlos
authors.initials: C
authors.lastname: López
authors.forename: Jorge
authors.initials: J
authors.lastname: Martín
authors.forename: Iris
authors.initials: I
authors.lastname: Valdés
authors.forename: Delfina
authors.initials: D
authors.lastname: del Rosario
authors.forename: Gerardo
authors.initials: G
authors.lastname: Guillén
authors.forename: María G
authors.initials: MG
authors.lastname: Guzmán
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000276:immunology*; D003715:Dengue / Q000517:prevention & control; D003716:Dengue Virus / Q000737:chemistry*; D004195:Disease Models, Animal; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology*; D014765:Viral Vaccines / Q000008:administration & dosage
chemical_list: D001425:Bacterial Outer Membrane Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; C112623:lpdA protein, Neisseria meningitidis