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In vitro evaluation of neuraminidase inhibitors using the neuraminidase-dependent release assay of hemagglutinin-pseudotyped viruses

Su, Ching-Yao, Wang, Shi-Yun, Shie, Jiun-Jie, Jeng, King-Song, Temperton, Nigel J., Fang, Jim-Min, Wong, Chi-Huey, Cheng, Yih-Shyun E. (2008) In vitro evaluation of neuraminidase inhibitors using the neuraminidase-dependent release assay of hemagglutinin-pseudotyped viruses. Antiviral Research, 79 (3). pp. 199-205. ISSN 0166-3542. (doi:10.1016/j.antiviral.2008.03.002) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided) (KAR id:29996)

The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided.
Official URL:
http://dx.doi.org/10.1016/j.antiviral.2008.03.002

Abstract

For the treatment of influenza virus infections, neuraminidase inhibitors (NAIs) that prevent the release of virus particles have been effective against most influenza strains. Several neuraminidase (NA) assays are available for the evaluation of NAIs. To understand the NAI functions under physiological conditions, assays mimicking viral particle release should be useful. We have constructed retrovirus-based reporter viruses that are pseudotyped with hemagglutinin (HA) glycoprotein by transfection of producer cells using plasmids expressing retroviral gag-pol, influenza HA, NA, and firefly luciferase genes. Similarly to the life cycle of influenza viruses, the release of pseudotype viruses also requires neuraminidase functions. This requirement was used to develop an assay to evaluate NAI activities by measuring inhibition of pseudotype virus production at different NAI concentrations. The pseudotype virus release assay was used to determine the IC(50) values of Oseltamivir carboxylate, Zanamivir, and the novel phosphonate congeners of Oseltamivir against N1 group neuraminidases and their H274Y Oseltamivir carboxylate-resistant mutants. The deduced IC(50) values obtained using the release assay correlated with those determined using the fluorogenic substrate 2'-(4-methylumbelliferyl)-alpha-d-N-acetylneuraminic acid (MUNANA) and also correlated with the infectivity results.

Item Type: Article
DOI/Identification number: 10.1016/j.antiviral.2008.03.002
Uncontrolled keywords: Antiviral, Drug resistance, Neuraminidase inhibitor, Pseudotype virus, Viral release assay, 2' (4 methylumbelliferyl) alpha dextro n acetylneuraminic acid, 4 acetamido 5 amino 3 (1 ethylpropoxy) 1 cyclohexene 1 carboxylic acid, Gag protein, hemagglutinin, Influenza virus hemagglutinin, luciferase, n acetylneuraminic acid, phosphonic acid derivative, sialidase, sialidase inhibitor, zanamivir, antiviral resistance, article, assay, concentration response, correlation analysis, drug potency, enzyme activity, enzyme assay, genetic transfection, IC 50, in vitro study, influenza, Influenza virus, luciferase gene, nonhuman, nucleotide sequence, plasmid, priority journal, protein expression, pseudotype virus, Retrovirus, viral release assay, virus gene, virus infectivity, virus inhibition, virus particle, virus strain, Animals, Antiviral Agents, Cell Line, Chick Embryo, Drug Evaluation, Preclinical, Enzyme Inhibitors, Genes, Reporter, Genetic Engineering, Hemagglutinins, Viral, Humans, Influenza, Human, Leukemia Virus, Murine, Neuraminidase, Orthomyxoviridae, Oseltamivir, Virus Shedding, Zanamivir, Orthomyxoviridae
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Divisions > Division of Natural Sciences > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 02 Aug 2012 10:37 UTC
Last Modified: 16 Nov 2021 10:07 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/29996 (The current URI for this page, for reference purposes)

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