A Prototype Recombinant Vaccine Against Respiratory Syncytial Virus and Parainfluenza Virus Type 3

• Published in: Bio/Technology 1994-08, Vol.12 (8), p.813-818
• Main Authors: Du, Run-Pan, Jackson, Gail E. D, Wyde, Philip R, Yan, Wei-Yao, Wang, Qijun, Gisonni, Lucy, Sanhueza, Sonia E, Klein, Michel H, Ewasyshyn, Mary E
• Format: Article
• Language: English
• Subjects: Agriculture; Amino Acid Sequence; Animals; Antibodies, Monoclonal - immunology; Baculoviridae - genetics; Base Sequence; Bioinformatics; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene Transfer Techniques; Genetic Engineering; Health. Pharmaceutical industry; Hemagglutinins - chemistry; Hemagglutinins - genetics; Hemagglutinins - immunology; HN Protein; Industrial applications and implications. Economical aspects; Life Sciences; Molecular Sequence Data; Moths - metabolism; Neuraminidase - chemistry; Neuraminidase - genetics; Neuraminidase - immunology; Parainfluenza Virus 3, Human - immunology; Production of active biomolecules; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - immunology; respiratory syncytial virus; Respiratory Syncytial Viruses - immunology; Vaccines, Synthetic; Vaccins; Viral Envelope Proteins; Viral Fusion Proteins - chemistry; Viral Fusion Proteins - genetics; Viral Fusion Proteins - immunology; Viral Proteins - chemistry; Viral Proteins - genetics; Viral Proteins - immunology; Viral Vaccines
• ISSN: 0733-222X; 1087-0156; 2331-3684; 1546-1696
• DOI: 10.1038/nbt0894-813

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 F RSV -HN PIV-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 μg 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.