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The human Transmembrane Protease Serine 2 is necessary for the production of Group 2 influenza A virus pseudotypes

Ferrara, Francesca, Molesti, Eleonora, Böttcher-Friebertshäuser, Eva, Cattoli, Giovanni, Corti, Davide, Scott, Simon D., Temperton, Nigel J. (2013) The human Transmembrane Protease Serine 2 is necessary for the production of Group 2 influenza A virus pseudotypes. Journal of Molecular and Genetic Medicine, 7 . pp. 309-314. ISSN 1747-0862. (doi:10.4172/1747-0862.1000055) (KAR id:33308)

Abstract

The monomer of influenza haemagglutinin is synthesized as a single polypeptide precursor that during maturation is cleaved by proteases into two active subunits. Other studies have demonstrated that the human Transmembrane Protease Serine 2 (TMPRSS2) can cleave the HA of human seasonal influenza viruses. Consequently, we have investigated the use of human Transmembrane Protease Serine 2 to produce high titre influenza haemmagglutinin (HA) lentiviral pseudotypes from Group 2 influenza viruses. Such pseudotypes represent powerful and safe tools to study viral entry and immune responses. Influenza pseudotype particles are obtained by co-transfecting human embryonic kidney HEK293T/17 cells using plasmids coding for the influenza HA, HIV gag-pol and a lentiviral vector incorporating firefly luciferase. However, in order to produce Group 2 pseudotypes, it was necessary to co-transfect a plasmid expressing the TMPRSS2 endoprotease, to achieve the necessary HA cleavage for infective particle generation. These lentiviral pseudotypes were shown to transduce HEK293T/17 cells with high efficiency. This demonstrates that TMPRSS2 is necessary for the functional activation, in vitro, of both the HA of human seasonal influenza and also other Group 2 HA influenza strains. Additionally, we show that the Group 2 influenza pseudotype particles could be used as surrogate antigens in neutralization assays and are efficiently neutralized by corresponding influenza reference sera. These data demonstrate that the pseudotype system is a powerful method for serological surveillance of a wide range of influenza viruses.

Item Type: Article
DOI/Identification number: 10.4172/1747-0862.1000055
Additional information: [1] Published online: 20 February 2013.
Uncontrolled keywords: Haemagglutinin, protease cleavage, antibody response, pseudotype serology
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Divisions > Division of Natural Sciences > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 05 Mar 2013 13:07 UTC
Last Modified: 09 Mar 2023 11:32 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/33308 (The current URI for this page, for reference purposes)

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