Pseudotyped Viruses for Influenza.
Joanne Marie M Del Rosario1, Kelly A S da Costa1, Nigel J Temperton2
1Viral Pseudotype Unit, Medway School of Pharmacy, University of Kent and Greenwich at Medway, Chatham, UK.
Advances in Experimental Medicine and Biology
|March 15, 2023
Summary
Researchers created a comprehensive library of influenza pseudotypes (PV) for all major influenza virus types and subtypes. These PVs, along with the AutoPlate web app, offer safer, faster, and more accessible methods for influenza serological assays and antibody screening.
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Influenza viruses pose a significant global health threat, necessitating robust methods for serological surveillance and vaccine development.
- Traditional influenza assays can be time-consuming, require specialized containment, and may not cover all relevant viral subtypes.
- Detecting neutralizing antibodies against diverse influenza hemagglutinin (HA), hemagglutinin-esterase fusion (HEF), and neuraminidase (NA) proteins is crucial for understanding immune responses.
Purpose of the Study:
- To develop and validate a comprehensive library of influenza pseudotypes (PV) for all influenza A, B, C, and D virus subtypes.
- To establish pseudotype microneutralization (pMN) and pseudotype enzyme-linked lectin assay (pELLA) methods for sensitive detection of neutralizing antibodies and neuraminidase inhibition (NI) titers.
- To introduce AutoPlate, a web application for efficient analysis of pMN and pELLA data, including calculation of half-maximal inhibitory concentrations (IC50).
Main Methods:
- Construction of a diverse pseudotype library including HA, HEF, and NA pseudotypes covering all influenza A (IAV) HA and NA subtypes, influenza B (IBV) lineages, and representative influenza C (ICV) and D (IDV) HEF types.
- Utilization of pseudotype microneutralization (pMN) assays for detecting neutralizing antibodies against HA/HEF proteins and pseudotype enzyme-linked lectin assays (pELLA) for measuring neuraminidase inhibition (NI) titers.
- Development and application of the AutoPlate web app for automated data analysis, including plotting inhibition curves and calculating IC50 values.
Main Results:
- Successful production of a comprehensive influenza pseudotype library encompassing HA, HEF, and NA proteins from IAV, IBV, ICV, and IDV.
- Demonstrated utility of pMN and pELLA assays as sensitive and specific tools for measuring neutralizing antibodies and NI titers against a wide range of influenza viruses.
- Validated the AutoPlate web app for rapid, efficient, and reproducible analysis of serological data, simplifying data interpretation.
Conclusions:
- Influenza pseudotypes serve as effective and safer substitutes for live viruses in serological and neutralization assays.
- The developed pseudotype library and associated assays provide a versatile platform for influenza research, vaccine immunogenicity assessment, and serosurveillance.
- The AutoPlate web app enhances the accessibility and efficiency of influenza serological data analysis, facilitating broader adoption in research laboratories worldwide.
Keywords:
AutoPlateELLAHemagglutininInfluenzaInfluenza immunityInfluenza vaccineNeuraminidaseNeutralizationPseudotypesMore Related Videos
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