title: A prototype recombinant vaccine against respiratory syncytial virus and parainfluenza virus type 3.
abstract: We have produced a genetically-engineered chimeric protein composed of the external domains of the respiratory syncytial virus (RSV) fusion (F) protein and the parainfluenza virus type 3 (PIV-3) hemagglutinin-neuraminidase (HN) protein in insect cells using the baculovirus expression system. The yield of the soluble chimeric FRSV-HNPIV-3 protein could be increased approximately 2-fold by using Trichoplasia ni (High Five) insect cells in place of Spodoptera frugiperda (Sf9) for expression. The chimeric protein, purified from the supernatant of baculovirus-infected High Five cells by immunoaffinity chromatography was correctly processed at the F2-F1 proteolytic cleavage site. Immunochemical analysis of the chimera with a panel of anti-F and anti-HN monoclonal antibodies suggested that the antigenicity of the major F and HN neutralization epitopes of the chimeric protein was preserved. Immunization of cotton rats with two 1 or 10 micrograms doses of the chimeric protein adsorbed to aluminum phosphate elicited strong PIV-3 specific HAI responses as well as PIV-3 and RSV specific neutralizing antibodies, and at either dose completely protected against challenge with live RSV and PIV-3.
authors.affiliation: Connaught Centre for Biotechnology Research, Willowdale, Ontario, Canada.
authors.forename: R P
authors.initials: RP
authors.lastname: Du
authors.forename: G E
authors.initials: GE
authors.lastname: Jackson
authors.forename: P R
authors.initials: PR
authors.lastname: Wyde
authors.forename: W Y
authors.initials: WY
authors.lastname: Yan
authors.forename: Q
authors.initials: Q
authors.lastname: Wang
authors.forename: L
authors.initials: L
authors.lastname: Gisonni
authors.forename: S E
authors.initials: SE
authors.lastname: Sanhueza
authors.forename: M H
authors.initials: MH
authors.lastname: Klein
authors.forename: M E
authors.initials: ME
authors.lastname: Ewasyshyn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D015707:HN Protein; D006388:Hemagglutinins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D009036:Moths / Q000378:metabolism; D009439:Neuraminidase / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D010224:Parainfluenza Virus 3, Human / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D012136:Respiratory Syncytial Viruses / Q000276:immunology*; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines
chemical_list: D000911:Antibodies, Monoclonal; D015707:HN Protein; D006388:Hemagglutinins; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D014764:Viral Proteins; D014765:Viral Vaccines; C116204:attachment protein G; D009439:Neuraminidase