The efficiency of simian foamy virus vector type-1 (SFV-1) in nondividing cells and in human PBLs.
Current retroviral vectors based on murine leukemia virus (MuLV) are unable to efficiently transduce nondividing cells. Lentiviruses, such as the human immunodeficiency virus 1 (HIV-1) are efficient at transducing nondividing, growth-arrested, and post-mitotic cells, but due to complex safety considerations, they may have limited potential for human clinical gene transfer. For this reason, alternatives to MuLV and HIV-1 vectors need to be explored. In this paper, we have found that simian foamy virus vector (SFV-1) containing a CMV-LacZ expression cassette is able to efficiently transduce multiple cell types of various species that include epithelial, lymphoid, and hematopoietic-derived human cell lines and fibroblast cell lines of several species. Previously it was reported that foamy virus replication is cell cycle dependent (P. D. Bieniasz, R. A. Weiss, and M. O. McClure, 1995. J. Virol. 69, 7295-7299). However, others studies demonstrated nuclear import of viral DNA in arrested cells (A. Saibi, F. Puvion-Dutilleul, M. Schmid, J. Peries, and H. d. The 1997. J. Virol. 71, 1155-1161). Here, we show efficient LacZ transduction by SFV-1 vectors in several chemically arrested cell lines and terminally differentiated human neurons. SFV-1 vector can transduce cell lines arrested in G1/S phase of the cell cycle by aphidicolin treatment with similar efficiencies to that of dividing cells. The terminally differentiated human neural cell line, NT2N, was transduced with 30-50% efficiency, corroborating our data obtained with the arrested cell lines. To further examine whether the SFV-1 vector can efficiently deliver a gene into clinically important cells for gene therapy, we transduced primary human peripheral blood cells (PBLs) in the presence and absence of phytohemagglutanin (PHA) stimulation. We observed 81% transduction efficiency in non-stimulated PBLs and 87% in PHA-stimulated PBLs with vector infection carried out twice in 8 hours intervals at a multiplicity of infection of 1. Together, these data indicate that SFV-1 based retroviral vectors may provide a safe, efficient alternative to current onco- and lentiviral vectors for gene transfer in cells from a broad spectrum of lineages across species boundaries.
Mergia
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Department of Pathobiology, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32611, USA. mergiaa@mail.vetmed.ufl.edu
Chari
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Kolson
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Goodenow
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Ciccarone
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D000818:Animals; D019556:COS Cells; D002455:Cell Division; D002460:Cell Line; D002461:Cell Line, Transformed; D002522:Chlorocebus aethiops; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D007763:Lac Operon; D007963:Leukocytes, Mononuclear / Q000166:cytology; D051379:Mice; D016092:Spumavirus / Q000235:genetics / Q000502:physiology*; D014407:Tumor Cells, Cultured