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Characterization of lentiviral pseudotypes with influenza H5N1 hemagglutinin and their performance in neutralization assays
- Source :
- Journal of Virological Methods. 165:305-310
- Publication Year :
- 2010
- Publisher :
- Elsevier BV, 2010.
-
Abstract
- Pseudotype reporter viruses are being used as safe, quantitative, and high-throughput tools for assessing antibody neutralization for many viruses, including influenza. However, characterization of pseudotypes containing influenza hemagglutinin (HA-pseudotypes) is needed before this system is widely adopted for evaluating neutralizing antibodies in sera following vaccination or infection. In this report HA-pseudotype stocks were analyzed for HA content, stability, and performance in neutralization assays under various conditions. HA-pseudotypes produced with HA genes of H5 strains representing clades 1, 2.2, and 2.3.4 consistently contain similar HA contents, and infectivity was not greatly affected by the purity of the HA-pseudotype preparations or variations in storage conditions. HA-pseudotype neutralization titers using a reference serum panel were also consistent across a wide range of dilutions of HA-pseudotype stocks and correlated well with results from microneutralization assays involving replicating influenza. Concentrated HA-pseudotypes were further shown to work well in hemagglutination inhibition assays. Finally, antisera elicited by genetically modified HA, with changes in the polybasic cleavage site that have been used in some H5 vaccines and reduce pathogenicity, gave identical neutralization titers against HA-pseudotypes with wild type or modified HA. These findings support continued development of HA-pseudotypes as a robust tool for analyzing sera in vaccine and serologic studies.
- Subjects :
- Antiserum
Infectivity
Sheep
Hemagglutination assay
Influenza A Virus, H5N1 Subtype
biology
Lentivirus
Orthomyxoviridae
Hemagglutinin Glycoproteins, Influenza Virus
Antibodies, Viral
biology.organism_classification
medicine.disease_cause
Antibodies, Neutralizing
Virology
Neutralization
Influenza A virus subtype H5N1
Microbiology
Titer
Neutralization Tests
Influenza, Human
medicine
Influenza A virus
Animals
Humans
Subjects
Details
- ISSN :
- 01660934
- Volume :
- 165
- Database :
- OpenAIRE
- Journal :
- Journal of Virological Methods
- Accession number :
- edsair.doi.dedup.....34a82d077a07fdc00a7352b7319e856f