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The human Transmembrane Protease Serine 2 is necessary for the production of Group 2 influenza A virus pseudotypes | OMICS International | Abstract
ISSN: 1747-0862

Journal of Molecular and Genetic Medicine
Open Access

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Research Article

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

Francesca Ferrara, Eleonora Molesti, Eva Böttcher-Friebertshäuser, Giovanni Cattoli¥, Davide Cortiψ, Simon D Scott* and Nigel J Temperton*

Viral Pseudotype Unit, Medway School of Pharmacy, University of Kent, Central Avenue, Chatham Maritime, ME4 4TB, UK

Institute of Virology, Philipps University Marburg, Hans-Meerwein-Str. 2, 35043 Marburg, Germany,

¥FAO-OIE and National Reference Laboratory for Newcastle Disease and Avian Influenza, Istituto Zooprofilattico delle Venezie, Viale dell’Università, 10 35020 Legnaro (PD), Italy,

ψInstitute for Research in Biomedicine, Via Vela 6, 6500 Bellinzona, Switzerland.

Corresponding Author:
Simon Scott
Email: [email protected]
Tel: +44 (0)1634 202957
Fax: +44 (0)1634 883927
Nigel Temperton
Email: [email protected]
Tel: +44 (0)1634 202957
Fax: +44 (0)1634 883927

Received date: 31 October 2012; Revised date: 19 February 2013; Accepted date: 20 February 2013; Published date: 20 February 2013

© Copyright The Author(s): First Published by Library Publishing Media. This is an open access article, published under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/ by-nc/2.5). This license permits non-commercial use, distribution and reproduction of the article, provided the original work is appropriately acknowledged with correct citation details.

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 other Group 2 HA influenza strains. Additionally, we show that the Group 2 influenza pseudotype particles can be used as surrogate antigens in neutralization assays and are efficiently neutralized by corresponding influenza virus reference sera. These data demonstrate that the viral pseudotype system is a powerful method for serological surveillance of a wide range of influenza viruses.

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