Release of cytokines and cytotoxic effector molecules in supernatant on co-culture of NK cells with melanoma cell lines
(A–F) NK cells were co-cultured with melanoma cell lines for 5h, E:T ratio of 1:1 and release of (A) IFNγ, (B) TNF, (C) perforin, (D) granzyme A, (E) granzyme B and (F) granulysin was measured in the supernatant. Cell lines are arranged in order of NK cytotoxic susceptibility. Each data point represents an individual donor and data are shown as mean ± SEM (n = 9, divided among 3 independent experiments). Statistical analysis was performed comparing no target condition vs. each individual cell line using one-way ANOVA. Below each bar graph, the corresponding heatmaps of the 9 individual UCB donors are shown. The individual donors are arranged in increasing cytotoxic capacity based on the geometric mean (gMean) of % cytotoxicity of each individual donor against all cell lines.
(G) Pearson correlation coefficient (r) between percentage of cytotoxicity vs. IFNγ, TNF, perforin, granzyme A, granzyme B or granulysin per cell line. Significance is shown as p < 0.05 ∗, <0.01 ∗∗, 0.001 ∗∗∗, 0.0001 ∗∗∗∗.