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Journal of Virology, August 2005, p. 10672-10677, Vol. 79, No. 16
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.16.10672-10677.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Generation of an Influenza A Virus Vector Expressing Biologically Active Human Interleukin-2 from the NS Gene Segment

Christian Kittel, Boris Ferko, Martina Kurz, Regina Voglauer, Sabine Sereinig, Julia Romanova, Gabriela Stiegler, Hermann Katinger, and Andrej Egorov*

Institute of Applied Microbiology, University of Natural Resources and Applied Life Sciences, Muthgasse 18B, A-1190 Vienna, Austria

Received 12 March 2005/ Accepted 4 May 2005

Engineering of the influenza A virus NS1 protein became an attractive approach to the development of influenza vaccine vectors since it can tolerate large inserts of foreign proteins. However, influenza virus vectors expressing long foreign sequences from the NS1 open reading frame (ORF) are usually replication deficient in animals due to the abrogation of their NS1 protein function. In this study, we describe a bicistronic expression strategy based on the insertion of an overlapping UAAUG stop-start codon cassette into the NS gene, allowing the reinitiation of translation of a foreign sequence. Although the expression level of green fluorescent protein (GFP) from the newly created reading frame was significantly lower than that obtained previously from an influenza virus vector expressing GFP from the NS1 ORF, the bicistronic vector appeared to be replication competent in mice and showed outstanding genetic stability. All viral isolates derived from mouse lungs at 10 days postinfection were still capable of expressing GFP in infected cells. Utilizing this bicistronic approach, we constructed another recombinant influenza virus, allowing the secretion of biologically active human interleukin-2 (IL-2). Although this virus also replicated to high titers in mouse lungs, it did not display any mortality rate in infected animals, in contrast to control viruses. Moreover, the IL-2-expressing virus showed an enhanced CD8+ response to viral antigens in mice after a single intranasal immunization. These results indicate that influenza viruses could be engineered for the expression of biologically active molecules such as cytokines for immune modulation purposes.

* Corresponding author. Mailing address: Institute of Applied Microbiology, Muthgasse 18B, A-1190 Vienna, Austria. Phone: (43/1) 36006-6594. Fax: (43/1) 36 97 615. E-mail: a.egorov{at}iam.boku.ac.at.

Journal of Virology, August 2005, p. 10672-10677, Vol. 79, No. 16
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.16.10672-10677.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Hatta, M., Kohlmeier, C. K., Hatta, Y., Ozawa, M., Kawaoka, Y. (2009). Region Required for Protein Expression from the Stop-Start Pentanucleotide in the M Gene of Influenza B Virus. J. Virol. 83: 5939-5942 [Abstract] [Full Text]  
  • Wressnigg, N., Shurygina, A. P., Wolff, T., Redlberger-Fritz, M., Popow-Kraupp, T., Muster, T., Egorov, A., Kittel, C. (2009). Influenza B mutant viruses with truncated NS1 proteins grow efficiently in Vero cells and are immunogenic in mice. J. Gen. Virol. 90: 366-374 [Abstract] [Full Text]  
  • Ferko, B., Kittel, C., Romanova, J., Sereinig, S., Katinger, H., Egorov, A. (2006). Live Attenuated Influenza Virus Expressing Human Interleukin-2 Reveals Increased Immunogenic Potential in Young and Aged Hosts. J. Virol. 80: 11621-11627 [Abstract] [Full Text]  


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