High-efficient lentiviral vector-mediated gene transfer into primary human NK cells

Federica Micucci, Alessandra Zingoni, Mario Piccoli, Luigi Frati, Angela Santoni, Ricciarda Galandrini

Research output: Contribution to journalArticle

19 Citations (Scopus)

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 1α (EF1α) 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. Conclusion: Results of our study demonstrate that NK cells can be efficiently transduced by lentiviral vectors.

Original languageEnglish
Pages (from-to)1344-1352
Number of pages9
JournalExperimental Hematology
Volume34
Issue number10
DOIs
Publication statusPublished - Oct 2006

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Natural Killer Cells
Genes
Transfection
Peptide Elongation Factor 1
Gene Transfer Techniques
Interleukin-12
Transgenes
Reporter Genes
Interleukin-2
Cultured Cells
Flow Cytometry
HIV
Cell Line
Infection

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

Cite this

High-efficient lentiviral vector-mediated gene transfer into primary human NK cells. / Micucci, Federica; Zingoni, Alessandra; Piccoli, Mario; Frati, Luigi; Santoni, Angela; Galandrini, Ricciarda.

In: Experimental Hematology, Vol. 34, No. 10, 10.2006, p. 1344-1352.

Research output: Contribution to journalArticle

Micucci, F, Zingoni, A, Piccoli, M, Frati, L, Santoni, A & Galandrini, R 2006, 'High-efficient lentiviral vector-mediated gene transfer into primary human NK cells', Experimental Hematology, vol. 34, no. 10, pp. 1344-1352. https://doi.org/10.1016/j.exphem.2006.06.001
Micucci, Federica ; Zingoni, Alessandra ; Piccoli, Mario ; Frati, Luigi ; Santoni, Angela ; Galandrini, Ricciarda. / High-efficient lentiviral vector-mediated gene transfer into primary human NK cells. In: Experimental Hematology. 2006 ; Vol. 34, No. 10. pp. 1344-1352.
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