Oncoretrovirus and lentivirus vectors pseudotyped with lymphocytic choriomeningitis virus glycoprotein: generation, concentration, and broad host range.
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.
Beyer
Winfried R
WR
Heinrich-Pette-Institut für Experimentelle Virologie und Immunologie an der Universität Hamburg, Germany. beyer@hpi.uni-hamburg.de
Westphal
Manfred
M
Ostertag
Wolfram
W
von Laer
Dorothee
D
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D001483:Base Sequence; D016466:CHO Cells; D002415:Cats; D002460:Cell Line; D002461:Cell Line, Transformed; D006224:Cricetinae; D004279:DNA, Viral; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology*; D005910:Glioma; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D020014:K562 Cells; D016086:Lentivirus / Q000235:genetics / Q000502:physiology*; D008217:Lymphocytic choriomeningitis virus / Q000235:genetics*; D008969:Molecular Sequence Data; D012190:Retroviridae / Q000235:genetics / Q000502:physiology*; D014407:Tumor Cells, Cultured; D014461:Ultracentrifugation / Q000379:methods; D014764:Viral Proteins / Q000235:genetics*
D000956:Antigens, Viral; D004279:DNA, Viral; D006023:Glycoproteins; D014764:Viral Proteins; C115679:glycoprotein peptide, Lymphocytic choriomeningitis virus