Abstract
Recent advances in cancer immunotherapy emphasize the need for an efficient method to genetically modify effector lymphocytes to express exogenous “synthetic” genes. NK cells represent 10–20% of total lymphocytes in the peripheral blood of humans and play an essential role in clearing infections and malignant cells. A significant number of NK cells express and utilize non-clonotypic receptors that recognize cognate ligands expressed on a broad spectrum of target cells. Thus, NK cells can be utilized as potent immunotherapeutic tools with fewer limitations. Considerable amount of progress in improving effector functions through genetic manipulations has been centered around T cells. However, a similar technological and translational exploration on NK cells is lacking. One major constrain is the significantly low efficiency of lentiviral-mediated gene transductions into primary human or mouse NK cells. We found that dextran, a branched glucan polysaccharide, significantly improves the transduction efficiency of human and mouse primary NK cells. This highly reproducible methodology offers an approach that can help to improve gene delivery into NK cells and thereby cancer immunotherapy.
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References
Vivier E, Tomasello E, Baratin M, Walzer T, Ugolini S (2008) Functions of natural killer cells. Nat Immunol 9(5):503–510
Zitvogel L, Terme M, Borg C, Trinchieri G (2006) Dendritic cell-NK cell cross-talk: regulation and physiopathology. Curr Top Microbiol Immunol 298:157–174
Abel AM, Yang C, Thakar MS, Malarkannan S (2018) Natural killer cells: development, maturation, and clinical utilization. Front Immunol 9:1869. https://doi.org/10.3389/fimmu.2018.01869
Raulet DH, Vance RE (2006) Self-tolerance of natural killer cells. Nat Rev Immunol 6(7):520–531
Schuster IS, Coudert JD, Andoniou CE, Degli-Esposti MA (2016) Natural regulators: NK cells as modulators of T cell immunity. Front Immunol 7:235. https://doi.org/10.3389/fimmu.2016.00235
Landoni E, Savoldo B (2018) Treating hematological malignancies with cell therapy: where are we now? Expert Opin Biol Ther 18(1):65–75. https://doi.org/10.1080/14712598.2018.1384810
Liu E, Tong Y, Dotti G, Shaim H, Savoldo B, Mukherjee M, Orange J, Wan X, Lu X, Reynolds A, Gagea M, Banerjee P, Cai R, Bdaiwi MH, Basar R, Muftuoglu M, Li L, Marin D, Wierda W, Keating M, Champlin R, Shpall E, Rezvani K (2018) Cord blood NK cells engineered to express IL-15 and a CD19-targeted CAR show long-term persistence and potent antitumor activity. Leukemia 32(2):520–531. https://doi.org/10.1038/leu.2017.226
Rezvani K, Rouce R, Liu E, Shpall E (2017) Engineering natural killer cells for cancer immunotherapy. Mol Ther 25(8):1769–1781. https://doi.org/10.1016/j.ymthe.2017.06.012
Zhang C, Oberoi P, Oelsner S, Waldmann A, Lindner A, Tonn T, Wels WS (2017) Chimeric antigen receptor-engineered NK-92 cells: an off-the-shelf cellular therapeutic for targeted elimination of cancer cells and induction of protective antitumor immunity. Front Immunol 8:533. https://doi.org/10.3389/fimmu.2017.00533
Regunathan J, Chen Y, Kutlesa S, Dai X, Bai L, Wen R, Wang D, Malarkannan S (2006) Differential and nonredundant roles of phospholipase Cgamma2 and phospholipase Cgamma1 in the terminal maturation of NK cells. J Immunol 177(8):5365–5376
Jiang K, Zhong B, Gilvary DL, Corliss BC, Vivier E, Hong-Geller E, Wei S, Djeu JY (2002) Syk regulation of phosphoinositide 3-kinase-dependent NK cell function. J Immunol 168(7):3155–3164
Maasho K, Marusina A, Reynolds NM, Coligan JE, Borrego F (2004) Efficient gene transfer into the human natural killer cell line, NKL, using the Amaxa nucleofection system. J Immunol Methods 284(1–2):133–140
Becknell B, Trotta R, Yu J, Ding W, Mao HC, Hughes T, Marburger T, Caligiuri MA (2005) Efficient infection of human natural killer cells with an EBV/retroviral hybrid vector. J Immunol Methods 296(1–2):115–123. https://doi.org/10.1016/j.jim.2004.11.012
Simmons A, Alberola-Ila J (2016) Retroviral transduction of T cells and T cell precursors. Methods Mol Biol 1323:99–108. https://doi.org/10.1007/978-1-4939-2809-5_8
Froelich S, Tai A, Wang P (2010) Lentiviral vectors for immune cells targeting. Immunopharmacol Immunotoxicol 32(2):208–218. https://doi.org/10.3109/08923970903420582
Denning W, Das S, Guo S, Xu J, Kappes JC, Hel Z (2013) Optimization of the transductional efficiency of lentiviral vectors: effect of sera and polycations. Mol Biotechnol 53(3):308–314. https://doi.org/10.1007/s12033-012-9528-5
Rajasekaran K, Kumar P, Schuldt KM, Peterson EJ, Vanhaesebroeck B, Dixit V, Thakar MS, Malarkannan S (2013) Signaling by Fyn-ADAP via the Carma1-Bcl-10-MAP3K7 signalosome exclusively regulates inflammatory cytokine production in NK cells. Nat Immunol 14(11):1127–1136
Acknowledgements
Funding Support: We thank Lucia Sammarco and her Lulu’s Lemonade Stand for inspiration, motivation, and support. This work was supported in part by Ann’s Hope Melanoma Foundation (S.M. and M.S.T.); NIH R01 AI102893 (S.M.) and NCI R01 CA179363 (S.M. and M.S.T.); HRHM Program of MACC Fund/Children’s Hospital of Wisconsin (S.M.), Nicholas Family Foundation (S.M.); Gardetto Family (S.M.); MCW-Cancer Center-Large Seed Grant (S.M. & M.S.T.); and MACC Fund/Children’s Hospital of Wisconsin (M.S.T. and S.M.).
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Nanbakhsh, A., Malarkannan, S. (2020). Dextran Enhances the Lentiviral Transduction Efficiency of Murine and Human Primary NK Cells. In: Katz, S., Rabinovich, P. (eds) Cell Reprogramming for Immunotherapy. Methods in Molecular Biology, vol 2097. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0203-4_7
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DOI: https://doi.org/10.1007/978-1-0716-0203-4_7
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