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Journal of Virology, February 2002, p. 1488-1495, Vol. 76, No. 3
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.3.1488-1495.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Oncoretrovirus and Lentivirus Vectors Pseudotyped with Lymphocytic Choriomeningitis Virus Glycoprotein: Generation, Concentration, and Broad Host Range

Winfried R. Beyer,1* Manfred Westphal,2 Wolfram Ostertag,1 and Dorothee von Laer3*

Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg,1 Neurochirurgische Klinik, Universitätskrankenhaus Eppendorf, D-20251 Hamburg,2 Georg-Speyer-Haus, 60596 Frankfurt, Germany3

Received 6 August 2001/ Accepted 29 October 2001

Lymphocytic choriomeningitis virus (LCMV) is a noncytopathic arenavirus shown to infect a broad range of different cell types. Here, we combined the beneficial characteristics of the LCMV glycoprotein (LCMV-GP) and those of retroviral vectors to generate a new, safe, and efficient gene transfer system. These LCMV-GP pseudotypes were systematically compared with vectors containing the widely used amphotropic murine leukemia virus envelope (A-MLVenv) or the vesicular stomatitis virus G protein (VSV-G). Production of LCMV-GP-pseudotyped oncoretroviral and lentiviral vectors by transient transfection resulted in vector titers similar to those with A-MLVenv or VSV-G. In contrast to A-MLVenv particles, LCMV-GP pseudotypes could be efficiently concentrated by ultracentrifugation without loss of vector titer. Unlike the cell-toxic VSV-G, a stable retroviral packaging cell line constitutively expressing LCMV-GP could be established. Vectors pseudotyped with LCMV-GP efficiently transduced many cell lines from different species and tissues relevant for gene therapy. Transduction of human glioma cells was studied in detail. These cells are a major target for cancer gene therapy and were transduced more efficiently with LCMV-GP-pseudotyped vectors than with the generally used A-MLVenv particles. The high stability, low toxicity, and broad host range make LCMV-GP-pseudotyped vectors attractive for gene transfer applications. The recombinant LCMV-GP-pseudotyped vectors will also allow functional characterization of naturally occurring and recombinant LCMV-GP variants.

* Corresponding author. Mailing address for Winfried R. Beyer: Heinrich-Pette-Institut, Martinistr. 52, D-20251 Hamburg, Germany. Phone: 49-40-48051-270. Fax: 49-40-48051-187. E-mail: beyer{at}hpi.uni-hamburg.de. Mailing address for Dorothee von Laer: Georg-Speyer-Haus, Paul-Ehrlich-Str. 42-44, D-60596 Frankfurt, Germany. Phone: 49-69-63395-232. Fax: 49-69-63395-297. E-mail: laer{at}em.uni-frankfurt.de.

Journal of Virology, February 2002, p. 1488-1495, Vol. 76, No. 3
0022-538X/01/$04.00+0     DOI: 10.1128/JVI.76.3.1488-1495.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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