Expression of IL-15 in NK cells results in rapid enrichment and selective cytotoxicity of gene-modified effectors that carry a tumor-specific antigen receptor
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Natural killer (NK) cells hold promise for adoptive cancer immunotherapy but are dependent on cytokines such as interleukin (IL)-2 for growth and cytotoxicity. Here, we investigated the consequences of ectopic expression of IL-15 in human NK cells. IL-2 and IL-15 belong to the common γ chain family of cytokines and have overlapping activities. Transduction of clinically applicable NK-92 cells with lentiviral vectors encoding human IL-15 resulted in predominantly intracellular expression of the cytokine, and STAT5 activation, proliferation and cytotoxicity of the producer cells in the absence of IL-2. Growth of non-transduced bystander cells was not supported, allowing rapid enrichment of gene-modified cells solely by IL-2 withdrawal. This was also the case upon transduction of NK-92 and NKL cells with a bicistronic lentiviral vector encoding IL-15 and a chimeric antigen receptor (CAR) targeting the pancarcinoma antigen EpCAM. Effector cells co-expressing CAR and IL-15 continued to proliferate in the absence of exogenous cytokines and displayed high and selective cell-killing activity against EpCAM-expressing breast carcinoma cells that were resistant to the natural cytotoxicity of unmodified NK cells. This strategy facilitates rapid isolation and continuous expansion of retargeted NK cells and may extend their potential clinical utility.
KeywordsInterleukin-15 Natural killer cells Lentiviral vector scFv antibody Chimeric antigen receptor EpCAM
This work was supported in part by grants from the Bundesministerium für Bildung und Forschung (BMBF) FKZ 01GU0805, the Deutsche Forschungsgemeinschaft (DFG) GRK1172, and the LOEWE Center for Cell and Gene Therapy Frankfurt (CGT). We thank Dr. Uwe Zangemeister-Wittke, University of Bern, for MOC31 antibody, Dr. Manuel Grez, Georg-Speyer-Haus, for helpful discussions and for providing lentiviral vectors and packaging constructs, Dr. Stefan Stein and Mr. Tevik Merovci, Georg-Speyer-Haus for help with flow cytometric cell sorting, and Mrs. Annemarie Schimpf, Georg-Speyer-Haus, for excellent technical assistance.
Conflict of interest
The authors declare that they have no conflict of interest.
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