After 14 days of culture, cell products became significantly enriched in NK cells (day 0 with mean 23.5%; range 5%-46% versus
day 14 with mean 80%; range 60%-95%, n = 6, P = 0.0001 data not shown). Expansion efficiency was comparable between PBMC derived from solid tumor patients versus healthy donor PBMC (mean 316 fold; range 1-1795 with n = 6 versus mean 165 fold; range 4-567 with n = 6, P = 0.6685). These data suggest that NK cells are efficiently expanded from PBMC from normal individuals and more importantly, from patients with various solid tumors without the need SC75741 price of primary enrichment protocols. NK cell expansion turns the receptor balance towards activation and Emricasan chemical structure results in autologous gastric tumor cell lysis Human NK cells maintain self tolerance by the expression of at least one inhibitory receptor specific for autologous HLA class I which prevents cytotoxicity against autologous cells [21]. To establish cytotoxicity against autologous target cells, inhibitory signals must be overcome, either by (i) down-regulation of inhibitory ligands on the XAV939 tumor cell, (ii) enhanced expression of activating receptors on NK cells, (iii) expression of ligands on the tumor target
that activate the NK cell or (iv) a combination of thereof. Since NK cell activation is affected by cytokines such as IL-2 and IL-15 [22], we sought to determine if NK cells expanded from PBMC were phenotypically different Evodiamine from non-expanded NK cells (Table 2). Table 2 Phenotypic changes on human NK cells after 14 days of expansion Healthy donors (n = 6) Patient 1 Patient 2 Day 0 Day 14 Mean (%) Range (%) Mean (%) Range (%) P-value a, b (%) Day 0 Day 14 Day 0 Day 14 Activating receptors DNAM-1 83 72-90 94 89-97 0.0335 (↑) 90 97 37 90 NKG2D 83 51-98 96 93-99 0.1074 30 94 87 98 NKp46 68 27-91 87 64-97 0.0161 (↑) 52 95 19 70 NKp44 3 2-5 59 16-93 0.0039 (↑) 0,3
29 0,4 15 NKp30 52 11-93 82 67-97 0.0131 (↑) 7 63 15 70 Inhibitory receptors KLRD1 68 56-82 92 86-95 0.0012 (↑) ND 98 49 95 NKG2A 46 14-67 68 34-89 0.00118 (↑) ND 84 7 8 KIR3DL1 22 10-37 29 17-38 0.1526 ND 21 5 3 KIR3DL2/3 28 9-48 29 14-44 0.7858 ND 35 88 96 LIR1 22 13-37 6 3-9 0.0142 (↓) ND 18 70 44 a Significant differences (P < 0.05) are indicated in bold b Arrows indicate significant increase (↑) or significant decrease (↓) ND; not determined In expanded NK cells from normal individuals, no significant change was observed in inhibitory receptors KIR3DL1 (P = 0.1526), KIR3DL2/3 (P = 0.7858) and the activating receptor NKG2D (P = 0.1074). In contrast, activating receptors DNAM-1 (P = 0.0061), NKp46 (P = 0.0161), NKp44 (P = 0.0039) and NKp30 (P = 0.0131) were significantly increased in expression after 14 days of expansion. Interestingly, KLRD1 (P = 0.0012) and NKG2A (P = 0.