A prototype recombinant vaccine against respiratory syncytial virus and parainfluenza virus type 3.
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.
Du
R P
RP
Connaught Centre for Biotechnology Research, Willowdale, Ontario, Canada.
Jackson
G E
GE
Wyde
P R
PR
Yan
W Y
WY
Wang
Q
Q
Gisonni
L
L
Sanhueza
S E
SE
Klein
M H
MH
Ewasyshyn
M E
ME
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
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