Establishment of retroviral pseudotypes with influenza hemagglutinins from H1, H3, and H5 subtypes for sensitive and specific detection of neutralizing antibodies.
Pseudotype reporter viruses provide a safe, quantitative, and high-throughput tool for assessing antibody neutralization for many viruses, including high pathogenicity H5 and H7 influenza A strains. However, adapting this system to other influenza subtypes has been hampered by variations in the protease cleavage site of hemagglutinin (HA) that make it less susceptible to the cleavage required for infectivity. In this report several proteases, reporter vectors, and cell substrates were evaluated while optimizing pseudovirus production, and robust methods were established for sensitive and specific neutralization of pseudotypes carrying influenza H1, H3, and H5 subtype HA that correlates well with titers obtained in microneutralization assays involving replicating influenza virus These findings should facilitate broad use of HA-pseudotypes that remove the need for infectious virus in a range of applications, including neutralization assays for serological surveys of viral infection and evaluations of vaccine sera.
Wang
Wei
W
Division of Viral Products, Center for Biologics Evaluation and Research, US Food and Drug Administration, 1401 Rockville Pike, Rockville, MD 20852, USA.
Butler
Eboneé N
EN
Veguilla
Vic
V
Vassell
Russell
R
Thomas
J Terrig
JT
Moos
Malcolm
M
Ye
Zhiping
Z
Hancock
Kathy
K
Weiss
Carol D
CD
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D002460:Cell Line; D005822:Genetic Vectors; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology; D053122:Influenza A Virus, H3N2 Subtype / Q000276:immunology; D053124:Influenza A Virus, H5N1 Subtype / Q000276:immunology; D009980:Influenza A virus / Q000145:classification* / Q000235:genetics / Q000276:immunology*; D009500:Neutralization Tests; D011817:Rabbits; D012190:Retroviridae / Q000235:genetics* / Q000472:pathogenicity; D012680:Sensitivity and Specificity
D000914:Antibodies, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus