title: Subunit influenza vaccine candidate based on CD154 fused to HAH5 increases the antibody titers and cellular immune response in chickens.
abstract: World Health Organization has a great concern about the spreading of avian influenza virus H5N1. To counteract its massive spread, poultry vaccination is highly recommended together with biosecurity measures. In our study, a recombinant vaccine candidate based on the fusion of extracellular segments of hemagglutinin (HA) H5 of avian influenza virus and chicken CD154 (HACD) is tested with the aim of enhancing humoral and cellular immune responses in chickens. Protein expression was carried out by transducing several mammalian cell lines with recombinant adenoviral vectors. HACD purification was assessed by three distinct purification protocols: immunoaffinity chromatography by elution at acidic pH or with a chaotropic agent and size exclusion chromatography. Humoral and cellular immune responses were measured using the hemagglutination inhibition assay and the semiquantitative real time PCR, respectively. The results showed that humoral response against HACD was significantly higher than the obtained with HA alone after booster (P<0.01, P<0.05). From HACD molecules purified by distinct protocols, only the obtained by size exclusion chromatography generated hemagglutinationin-inhibition activity. IFN-γ levels indicated that cellular immune response was significantly higher with HACD, in its pure or impure form, compared to its counterpart HA (P<0.01). These data demonstrate that HACD is able to significantly enhance humoral and cellular immune responses against HA antigen, which make this fusion protein a promising subunit vaccine candidate against H5N1 virus outbreaks.
authors.affiliation: Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), P.O. Box 6162, Havana 10600, Cuba. alain.pose@cigb.edu.cu
authors.forename: Alaín González
authors.initials: AG
authors.lastname: Pose
authors.forename: Julia Noda
authors.initials: JN
authors.lastname: Gómez
authors.forename: Alina Venereo
authors.initials: AV
authors.lastname: Sánchez
authors.forename: Armando Vega
authors.initials: AV
authors.lastname: Redondo
authors.forename: Elsa Rodríguez
authors.initials: ER
authors.lastname: Rodríguez
authors.forename: Raquel Montesino
authors.initials: RM
authors.lastname: Seguí
authors.forename: Ernesto Manuel González
authors.initials: EM
authors.lastname: Ramos
authors.forename: María Pilar Rodríguez
authors.initials: MP
authors.lastname: Moltó
authors.forename: Elaine Santana
authors.initials: ES
authors.lastname: Rodríguez
authors.forename: Liliam Rios
authors.initials: LR
authors.lastname: Cordero
authors.forename: Alina Rodríguez
authors.initials: AR
authors.lastname: Mallón
authors.forename: Carlos Borroto
authors.initials: CB
authors.lastname: Nordelo
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000235:genetics / Q000276:immunology; D023201:CD40 Ligand / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D002645:Chickens; D006385:Hemagglutination Inhibition Tests; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism*; D006388:Hemagglutinins / Q000235:genetics / Q000276:immunology; D006801:Humans; D007111:Immunity, Cellular; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology*; D007252:Influenza Vaccines / Q000235:genetics / Q000276:immunology*; D005585:Influenza in Birds / Q000276:immunology* / Q000517:prevention & control* / Q000821:virology; D022223:Vaccines, Subunit / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D006388:Hemagglutinins; D007252:Influenza Vaccines; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; C109963:hemagglutinin, avian influenza A virus; D023201:CD40 Ligand