What is it about?
It is a comprehensive review article on the field of influenza pseudotypes and their multiple applications in gene therapy, serology, sero-surveillance, restriction factors, antiviral screening, immunogenicity testing and vaccination. Infographics throughout to add clarity.
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Why is it important?
Lentiviral and other vectors are gaining increasing importance not only for gene therapy, but also as a tool for the production of surrogate viruses for use in neutralization assays. This review extracts all of the production, titration and neutralization assay methodologies from over 100 publications and enables consensus protocols to be proposed.
Perspectives
This review is a very comprehensive overview of the primary literature concerning the production and application of influenza pseudotypes. It also provides consensus protocols for production, titration and neutralisation assays that were informed from the peer reviewed literature.
Professor Nigel James Temperton
University of Kent
Read the Original
This page is a summary of: Pseudotype-Based Neutralization Assays for Influenza: A Systematic Analysis, Frontiers in Immunology, April 2015, Frontiers,
DOI: 10.3389/fimmu.2015.00161.
You can read the full text:
Resources
Consensus protocol guide
Consensus protocols for the production, titration and neutralisation of influenza pseudotypes - a handy printable guide
Influenza pseudotype poster
Poster on the use of influenza pseudotypes in pandemic preparedness.
Pseudotype neutralisation set up guide
Pseudotype neutralization assays are powerful tools to study functional antibody responses against viruses in low biosafety laboratories. However, protocols described in the literature differ widely with respect to material, reagents, and methods used to perform these assays and to analyse the raw data generated. This could result in discrepancies between the results of different laboratories even when the same pseudotypes and the same samples are analysed. Here, we describe, in detail, an experimental protocol to perform pseudotype neutralization assays using lentiviral pseudotypes bearing influenza haemagglutinin and expressing firefly luciferase. We also present the steps necessary to analyse the data and calculate the half maximal inhibitory concentration of the sera analysed. This protocol will provide support for the validation and the standardization of the pseudotype neutralization assay for influenza virus serology. Additionally, it will provide a starting point for the development of pseudotype neutralization assays using pseudotypes bearing other viral envelope proteins.
Contributors
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