Hemagglutinin-pseudotyped green fluorescent protein-expressing influenza viruses for the detection of influenza virus neutralizing antibodies.
Influenza virus is a highly contagious virus that causes yearly epidemics and occasional pandemics of great consequence. Influenza virus neutralizing antibodies (NAbs) are promising prophylactic and therapeutic reagents. Detection of NAbs in serum samples is critical to evaluate the prevalence and spread of new virus strains. Here we describe the development of a simple, sensitive, specific, and safe screening assay for the rapid detection of NAbs against highly pathogenic influenza viruses under biosafety level 2 (BSL-2) conditions. This assay is based on the use of influenza viruses in which the hemagglutinin (HA) gene is replaced by a gene expressing green fluorescent protein (GFP). These GFP-expressing influenza viruses replicate to high titers in HA-expressing cell lines, but in non-HA-expressing cells, their replication is restricted to a single cycle.
Martínez-Sobrido
Luis
L
Department of Microbiology, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, NY 10029, USA.
Cadagan
Richard
R
Steel
John
J
Basler
Christopher F
CF
Palese
Peter
P
Moran
Thomas M
TM
García-Sastre
Adolfo
A
D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood*; D002460:Cell Line; D004196:Disease Outbreaks; D004285:Dogs; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000276:immunology*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D009980:Influenza A virus / Q000235:genetics* / Q000276:immunology* / Q000472:pathogenicity; D007251:Influenza, Human / Q000453:epidemiology / Q000276:immunology / Q000821:virology; D009500:Neutralization Tests / Q000379:methods* / Q000706:statistics & numerical data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins