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. 2005 Jan;296(1-2):115-23.
doi: 10.1016/j.jim.2004.11.012. Epub 2004 Dec 8.

Efficient infection of human natural killer cells with an EBV/retroviral hybrid vector

Brian Becknell  1 , Rossana Trotta, Jianhua Yu, Wei Ding, Hsiaoyin C Mao, Tiffany Hughes, Trent Marburger, Michael A Caligiuri
Affiliations

Efficient infection of human natural killer cells with an EBV/retroviral hybrid vector

Brian Becknell et al. J Immunol Methods. 2005 Jan.

Abstract

Molecular characterization of human natural killer (NK) cells will require targeted gene delivery to inhibit and activate specific signaling pathways, yet to our knowledge, an effective means to deliver such products for long-term gene expression without disrupting normal cellular processes has not been described. In this study, we have developed a retroviral strategy to effectively express gene products in the NK cell, whereby its effector functions of cytotoxicity and cytokine production remain intact. Using an EBV/retroviral hybrid vector, we demonstrate infection of human peripheral blood NK cells with simultaneous expression of a marker for infection--the enhanced green fluorescent protein (EGFP)--along with various genes of interest. This technique results in successful infection of the CD56dim NK population that predominates among human peripheral blood NK and is the effector of antibody-dependent cellular cytotoxicity and natural killing. In addition, we demonstrate infection of the CD56bright NK subset as well as the NK-92 cell line. In summary, we have devised an efficient and reproducible methodology for the targeted delivery of gene products to human NK cells that should now provide opportunities to dissect the molecular processes critical to normal NK cell physiology.

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