Characterization of baculovirus-expressed hemagglutinin and fusion glycoproteins of the attenuated measles virus strain AIK-C.
With measles virus cDNA of the avirulent vaccine strain AIK-C, two cDNAs of H or F genes were amplified by the polymerase chain reaction. The amplified cDNAs were inserted respectively to the baculovirus transfer vector pAcYM1 derived from the nuclear polyhedrosis virus of Autographa californica (AcNPV). After co-transfection of the transfer vectors with AcNPV DNA to Spodoptera frugiperda cells, recombinant baculoviruses were screened by plaque assay, and the viruses containing H-cDNA or F-cDNA were named H-AIK or F-AIK, respectively. By Western blot analyses, the band around 80 kDa and some smaller bands were appeared in the H-AIK infected S. frugiperda cells, and the band around 40 kDa was detected in the F-AIK infected cells. Immunofluorescence studies on unfixed S. frugiperda cells infected with H- or F-AIK recombinants showed that both antigens were transported to the cell surface. When green monkey red blood cells were added to the recombinant infected cells, H-AIK infected cells showed haemadsorption, and cells infected with F-AIK lysed the red blood cells. The recombinant proteins elicited the neutralizing antibodies against measles virus.
Takehara
K
K
Laboratory of Poultry Diseases, School of Veterinary Medicine and Animal Sciences, Kitasato University, Aomori, Japan.
Hashimoto
H
H
Ri
T
T
Mori
T
T
Yoshimura
M
M
D000818:Animals; D000914:Antibodies, Viral; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006389:Hemagglutinins, Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D008459:Measles virus / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D008969:Molecular Sequence Data; D009036:Moths; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014760:Viral Fusion Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D014774:Virulence / Q000235:genetics
D000914:Antibodies, Viral; D004279:DNA, Viral; D006389:Hemagglutinins, Viral; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins