title: Efficient infection of human natural killer cells with an EBV/retroviral hybrid vector.
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.
authors.affiliation: Medical Scientist Program, Integrated Biomedical Graduate Program, College of Medicine and Public Health, The Ohio State University, Columbus, OH 43210, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Becknell
authors.forename: Rossana
authors.initials: R
authors.lastname: Trotta
authors.forename: Jianhua
authors.initials: J
authors.lastname: Yu
authors.forename: Wei
authors.initials: W
authors.lastname: Ding
authors.forename: Hsiaoyin C
authors.initials: HC
authors.lastname: Mao
authors.forename: Tiffany
authors.initials: T
authors.lastname: Hughes
authors.forename: Trent
authors.initials: T
authors.lastname: Marburger
authors.forename: Michael A
authors.initials: MA
authors.lastname: Caligiuri
mesh_terms: D019002:CD56 Antigen / Q000276:immunology; D002478:Cells, Cultured; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics*; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics*
chemical_list: D019002:CD56 Antigen; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma