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Journal of Virology, March 2003, p. 3851-3858, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3851-3858.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Chimeric and Pseudotyped Parvoviruses Minimize the Contamination of Recombinant Stocks with Replication-Competent Viruses and Identify a DNA Sequence That Restricts Parvovirus H-1 in Mouse Cells

Claudia Wrzesinski, Lia Tesfay, Nathalie Salomé, Jean-Claude Jauniaux, Jean Rommelaere, Jan Cornelis, and Christiane Dinsart*

Applied Tumor Virology, Abteilung F0100 and INSERM U375, Deutsches Krebsforschungszentrum, Heidelberg, Germany

Received 31 July 2002/ Accepted 23 December 2002

Recent studies demonstrated the ability of the recombinant autonomous parvoviruses MVMp (fibrotropic variant of the minute virus of mice) and H-1 to transduce therapeutic genes in tumor cells. However, recombinant vector stocks are contaminated by replication-competent viruses (RCVs) generated during the production procedure. To reduce the levels of RCVs, chimeric recombinant vector genomes were designed by replacing the right-hand region of H-1 virus DNA with that of the closely related MVMp virus DNA and conversely. Recombinant H-1 and MVMp virus pseudotypes were also produced with this aim. In both cases, the levels of RCVs contaminating the virus stocks were considerably reduced (virus was not detected in pseudotyped virus stocks, even after two amplification steps), while the yields of vector viruses produced were not affected. H-1 virus could be distinguished from MVMp virus by its restriction in mouse cells at an early stage of infection prior to detectable viral DNA replication and gene expression. The analysis of the composite viruses showed that this restriction could be assigned to a specific genomic determinant(s). Unlike MVMp virus, H-1 virus capsids were found to be a major determinant of the greater permissiveness of various human cell lines for this virus.

* Corresponding author. Mailing address: Applied Tumor Virology, Abteilung F0100 and INSERM U375, Deutsches Krebsforschungszentrum, Postfach 101949, D-69009 Heidelberg, Germany. Phone: 49 6221 424965. Fax: 49 6221 424962. E-mail: c.dinsart{at}dkfz.de.

Journal of Virology, March 2003, p. 3851-3858, Vol. 77, No. 6
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.6.3851-3858.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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