title: High-efficient lentiviral vector-mediated gene transfer into primary human NK cells.
abstract: OBJECTIVE
  The long-term transfection of genes into primary natural killer (NK) cells without disrupting normal cellular functions has been proven to be difficult with currently available gene-transfer methods. In this study, we establish a lentiviral vector-based technique for improved gene transfer into human NK cells in vitro and we report on high-efficient transduction of freshly isolated as well as cultured primary NK cells.
  
  
  METHODS
  Freshly isolated or primary cultured human NK cells, as well as the human NK cell line YTS, were transduced with replication-incompetent human immunodeficiency virus (HIV)-based lentiviral vector bearing a GFP reporter gene or a gene of interest under the control of the elongation factor 1alpha (EF1alpha) promoter. Transduction efficiencies were monitored by flow cytometry.
  
  
  RESULTS
  A long-term transgene expression was detected in up to 98% of YTS NK cells, whereas in freshly isolated or primary cultured NK cells exposed to interleukin (IL)-2 plus IL-12 upon infection, efficiency was in the range of 50% to 90%. Moreover, in freshly isolated quiescent NK cells a transfection efficiency of 18% to 20% was achieved without stimulation. Notably, no major phenotypic and functional modifications were observed in transduced cells with respect to control cells: the expression levels of activating receptors, CD69-antigen induction as well as cytotoxic function were unaffected.
  
  
  CONCLUSIONS
  Results of our study demonstrate that NK cells can be efficiently transduced by lentiviral vectors.
authors.affiliation: Department of Experimental Medicine and Pathology, Istituto Pasteur-Fondazione Cenci-Bolognetti, University "La Sapienza," Rome, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Micucci
authors.forename: Alessandra
authors.initials: A
authors.lastname: Zingoni
authors.forename: Mario
authors.initials: M
authors.lastname: Piccoli
authors.forename: Luigi
authors.initials: L
authors.lastname: Frati
authors.forename: Angela
authors.initials: A
authors.lastname: Santoni
authors.forename: Ricciarda
authors.initials: R
authors.lastname: Galandrini
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000096:biosynthesis; D002478:Cells, Cultured; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D018664:Interleukin-12 / Q000494:pharmacology; D007694:Killer Cells, Natural / Q000166:cytology / Q000378:metabolism; D037181:Lectins, C-Type; D016086:Lentivirus; D008213:Lymphocyte Activation / Q000187:drug effects / Q000235:genetics; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D013997:Time Factors; D014161:Transduction, Genetic / Q000379:methods; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D018664:Interleukin-12; D037181:Lectins, C-Type; D020648:Peptide Elongation Factor 1; D000099192:CD69 Antigens