Inhibition of intracellular antiviral defense mechanisms augments lentiviral transduction of human natural killer cells: implications for gene therapy.
Adoptive immunotherapy with genetically modified natural killer (NK) cells is a promising approach for cancer treatment. Yet, optimization of highly efficient and clinically applicable gene transfer protocols for NK cells still presents a challenge. In this study, we aimed at identifying conditions under which optimum lentiviral gene transfer to NK cells can be achieved. Our results demonstrate that stimulation of NK cells with interleukin (IL)-2 and IL-21 supports efficient transduction using a VSV-G pseudotyped lentiviral vector. Moreover, we have identified that inhibition of innate immune receptor signaling greatly enhances transduction efficiency. We were able to boost the efficiency of lentiviral genetic modification on average 3.8-fold using BX795, an inhibitor of the TBK1/IKKɛ complex acting downstream of RIG-I, MDA-5, and TLR3. We have also observed that the use of BX795 enhances lentiviral transduction efficiency in a number of human and mouse cell lines, indicating a broadly applicable, practical, and safe approach that has the potential of being applicable to various gene therapy protocols.
Center for Hematology and Regenerative Medicine, Department of Medicine, Karolinska University Hospital Huddinge, Karolinska Institutet, SE-14186 Stockholm, Sweden.
Tolga
T
Sutlu
Sanna
S
Nyström
Mari
M
Gilljam
Birgitta
B
Stellan
Steven E
SE
Applequist
Evren
E
Alici
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D016207:Cytokines / Q000494:pharmacology; D015316:Genetic Therapy; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects; D042541:Intracellular Space / Q000187:drug effects / Q000821:virology*; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology* / Q000821:virology*; D016086:Lentivirus / Q000187:drug effects* / Q000235:genetics; D051379:Mice; D010641:Phenotype; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D051192:Receptors, Pattern Recognition / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D013997:Time Factors; D014161:Transduction, Genetic
D000998:Antiviral Agents; D016207:Cytokines; D047428:Protein Kinase Inhibitors; D051192:Receptors, Pattern Recognition