Stable transduction of primary human monocytes by simian lentiviral vector PBj.
Despite the ability to infect nonproliferating cells, current lentiviral vectors are inefficient at mediating gene transfer into quiescent primary human cells such as monocytes. Here, a replication-incompetent vector based on a molecular clone of simian immunodeficiency virus strain PBj (SIVsmmPBj1.9) was generated that, in contrast to lenti- and gamma-retroviral control vectors, enabled transfer of heterologous genes into human diploid fibroblasts and cell lines blocked in the G(0) phase of the cell cycle. Moreover, freshly isolated human monocytes refractory to HIV-1-derived vectors were efficiently transduced by the PBj vector independent of the viral Nef protein. Stable chromosomal integration of PBj-derived viral expression vectors was verified in transduced cells. The capability of the PBj vector to transduce quiescent cells such as unstimulated primary human monocytes is an important extension of human gene therapy perspectives.
Mühlebach
Michael D
MD
Division of Medical Biotechnology, Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, D-63225 Langen, Germany.
Wolfrum
Nina
N
Schüle
Silke
S
Tschulena
Ulrich
U
Sanzenbacher
Ralf
R
Flory
Egbert
E
Cichutek
Klaus
K
Schweizer
Matthias
M
D015139:Blotting, Southern; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002875:Chromosomes / Q000378:metabolism; D046508:Culture Techniques; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D005697:Galactosides / Q000494:pharmacology; D015693:Gene Products, nef / Q000378:metabolism; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D016086:Lentivirus / Q000235:genetics*; D008957:Models, Genetic; D009000:Monocytes / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D014161:Transduction, Genetic
D005697:Galactosides; D015693:Gene Products, nef; D007211:Indoles; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside