Expansion and Activation of Human Natural Killer Cells ex vivo in the Presence of Transgenic Feeder Cells
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nsion and Activation of Human Natural Killer Cells ex vivo in the Presence of Transgenic Feeder Cells E. P. Vashkevicha, A. A. Migasa, *, A. N. Meleshkoa, M. A. Matveenkoa, N. V. Strushkevichb, and T. V. Shmana aRepublican b
Scientific and Practical Center of Children’s Oncology and Hematology, Minsk, 223053 Belarus Skolkovo Institute of Science and Technology, Skolkovo Innovation Center Territory, Moscow, 143026 Russia *e-mail: [email protected] Received December 18, 2019; revised January 13, 2020; accepted January 15, 2020
Abstract—Preliminary results of clinical trials show that the use of activated and expanded ex vivo natural killer cells is a promising tool for adoptive immunotherapy of acute myeloid leukemia, as well as a number of solid tumors, when used together with targeted monoclonal antibodies. However, the introduction of this approach into clinical practice is limited by the possibility of obtaining a sufficient amount of the NK cell product with required characteristics. To solve this problem, we obtained transgenic feeder cells based on immortalized K562 cells expressing the recombinant membrane-bound variant of human interleukin-21 and 4-1BBL protein. Cocultivation of mononuclear cells obtained from the peripheral blood of ten healthy donors with genetically modified feeder cells resulted in significant expansion of natural killer cells (median expansion fold 21589 times; minimum, 3150 times; maximum, 304328 times) with a minimum content of Tcells (median, 0.5%; minimum, 0.06%; maximum, 4.7%) compared with their initial number. NK cells obtained in this way were not contaminated with the BCR–ABL1 oncogene from feeder cells and had a lower expression of c-MYC protooncogene compared to the initial level. Keywords: natural killer cells, ex vivo expansion and activation, adoptive antitumor immunotherapy, genetically engineered feeder cells DOI: 10.1134/S1990519X20050090
INTRODUCTION Natural killer (NK) cells are a population of lymphocytes of the innate immune system that are actively involved in the formation of the antitumor and antiviral immune response (Kiessling et al., 1975; Pross et al., 1975; Timonen et al., 1980). They are differentiated from the common precursor of lymphocytes in bone marrow. Further maturation and functional activity are observed mainly in peripheral blood and lymph nodes (Kondo et al., 1997; Carotta et al., 2011; Fathman et al., 2011; Sojka et al., 2014; Constantinides et al., 2015). The high cytotoxic activity of NK cells in relation to malignant cells of various origins makes them useful in the adoptive therapy of cancer. The effective application of this technology requires to have a sufficient amount of cellular product, characterized by the necessary purity and activation of NK cells. Abbreviations: NK—natural killer cell, NKT—NK-like T-cell, IL—interleukin, mIL—membrane-bound recombinant variant of interleukin, cDNA—complementary DNA synthesized on an RNA template, FC—feeder cell, MNC—mononuclear cell, mRNA—messenger RNA, PCR—polymerase chain reaction.
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