Expression of influenza A and B virus nucleoprotein antigens in baculovirus.
Full-length cDNA clones of the nucleoprotein (NP) genes of influenza A/Ann Arbor/6/60 and B/Ann Arbor/1/86 viruses were constructed from virion RNA and subsequently expressed in Spodoptera frugiperda (Sf9) cells using the baculovirus vector, Autographa californica nuclear polyhedrosis virus. Western blot analysis of lysates prepared from Sf9 cells infected with the recombinant viruses confirmed that the baculovirus-expressed NP antigens were reactive with monoclonal antibodies specific for either type A or B NP and with anti-NP antibodies in human serum samples. Electrophoretic analysis indicated that the expressed NP antigens comigrated with NP purified from influenza A or B virions and that the recombinant NP antigens represented greater than 10% of total protein in infected cells. Dilutions of clarified Sf9 cell lysates were used as antigens in a standard enzyme immunoassay to detect serum antibody specific for influenza A or B viruses. The results from assays using the baculovirus-expressed NP antigens showed good correlation with the results obtained using bacterially expressed NP antigen as well as complement fixation. Therefore, baculovirus-expressed NP antigens have the potential to be used to develop reproducible and routine assays for the serodiagnosis of influenza virus infections as an alternative to the complement fixation or haemagglutination inhibition tests.
Division of Viral and Rickettsial Diseases, Centers for Disease Control, Atlanta, Georgia 30333.
P A
PA
Rota
R A
RA
Black
B K
BK
De
M W
MW
Harmon
A P
AP
Kendal
D000328:Adult; D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics* / Q000276:immunology; D015153:Blotting, Western; D002460:Cell Line; D002648:Child; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000276:immunology; D009981:Influenza B virus / Q000235:genetics* / Q000276:immunology; D007304:Insect Viruses / Q000235:genetics*; D009036:Moths; D000077183:Occlusion Body Matrix Proteins; D012261:Ribonucleoproteins / Q000235:genetics* / Q000276:immunology; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins; D014771:Virion / Q000235:genetics
D000914:Antibodies, Viral; D000956:Antigens, Viral; D004279:DNA, Viral; D000077183:Occlusion Body Matrix Proteins; D012261:Ribonucleoproteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus