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Fig. 3 | Journal for ImmunoTherapy of Cancer

Fig. 3

From: Discovery of a novel natural killer cell line with distinct immunostimulatory and proliferative potential as an alternative platform for cancer immunotherapy

Fig. 3

CD56dimCD62L+ NK-like features of NK101. a NK101, NK-92 and primary human peripheral blood mononuclear cells (PBMCs) were stained with fluorochrome-conjugated anti-CD3, −CD19, −CD56, and -CD62L antibodies. Results are displayed as one-color histogram (left) or two-color contour (right) plots after gating live, CD3, CD19, and CD56+ cells. Red, blue and black indicate NK101, NK-92, primary CD56+ NK cells, respectively. In the histogram plot, gray-filled line represents isotype control and the numbers in the parenthesis represent geometric mean fluorescence intensity of CD56 (left). Results are representative of 3 independent experiments. b Cultured NK101 cells were treated with 10 ng/ml of indicated cytokines (except IL-2; 500 IU/ml). Cell expansion was assessed by MTS assay after 3 days (left) and IFN-γ secretion was measured by ELISA after 24 h (right). Data represent mean ± SD of triplicate wells from 3 independent experiments. **p < 0.01 versus the corresponding untreated groups. c The secretion of indicated cytokines or chemokines from rested, K562-(E:T = 4:1), or THP-1-(E:T = 4:1) co-cultured NK101 cells was measured by a multiplex immunoassay. Data represent mean ± SD of triplicate wells from 2 independent experiments. **p < 0.01 versus rested NK101. d NK101 cells were co-cultured with CFSE-labeled K562 or THP-1 cells at an effector-to-target ratio of 4:1 for 24 h in the absence (left) or presence of indicated neutralizing antibody (10 μg/ml) (right). Harvested cells were stained with Annexin-V and fixable viability dye, and CFSE+ tumor cells were analyzed by flow cytometry. Representative plots from 3 independent experiments are shown (left). Bar graphs represent mean ± SD of triplicate wells from 3 independent experiments (right). *p < 0.05, **p < 0.01 versus corresponding isotype control groups

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