A New Ex Vivo Method for Effective Expansion and Activation of Human Natural Killer Cells for Anti-Tumor Immunotherapy

Abstract

Preserving the activities of natural killer (NK) cells in human peripheral blood mononuclear cells (PBMCs) after ex vivo expansion and activation is critical for NK cell-based therapy. Collected from human PBMCs, the NK cells were expanded and activated. The expressions of surface receptors, cytotoxicity against tumor cells, and antibody-dependent cell-mediated cytotoxicity (ADCC) of the NK cells before and after expansion and activation were, respectively, compared. After expansion, the ADCC activity of healthy human NK cells was improved by 32 %, and the cytotoxicity against four types of tumor cells was increased by 19, 29, 26, and 28 %, respectively. The positive expression rates for the activating receptors NKG2D, CD94, NKp46, NKp30, and NKp44 of healthy human NK cells expanded ex vivo were increased by 60, 40, 20, 40, and 63 %, respectively, whereas those for the inhibitory receptors CD158b, NKB1, and NKAT showed no significant changes. The addition of an immunologically active peptide, "TKD," during cell expansion further increased NK cytotoxicity by approximately 10 %. The expanded and activated NK cells from cancer patients achieved average purity which was greater than 90 %, and the cytotoxicity against K562 cells was increased by more than 17 %. Compared with resting NK cells, NK cells both from healthy volunteers and cancer patients expanded and activated ex vivo using our method were significantly more active and demonstrated significantly increased anti-tumor activity. This method could be therefore used as a new and effective approach to meet requirements for anti-tumor immunotherapy.

Keywords: Activating receptor; Antibody-dependent cell-mediated cytotoxicity (ADCC); Ex vivo expansion; Inhibitory receptor; Natural killer cells.