Figure 3.

Intracellular levels of cytokines and cytotoxic effector molecules on co-culture of NK cells with melanoma cell lines

(A–F) NK cells were co-cultured with melanoma cell lines for 5h at the E:T ratio of 1:1 and percentage of (A) IFNγ⁺ cells, (B) TNF⁺ cells and gMFI of (C) perforin and (D) granzyme B was determined. Cell lines are arranged from high to low susceptibility. Red dashed line indicates baseline level of no target condition. 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.

(E) Pearson correlation coefficient (r) of percentage of cytotoxicity vs. % degranulation, % IFNγ⁺ cells, % TNF⁺ cells, gMFI of perforin or gMFI of granzyme B of effector + target (NK + tumor) conditions per cell line and data of all cell lines.

(F) Pearson correlation coefficient (r) of % cytotoxicity vs. intrinsic levels of perforin or granzyme B per cell line and gMean of all cell lines. Significance is shown as p < 0.05 ∗, <0.01 ∗∗, 0.001 ∗∗∗, 0.0001 ∗∗∗∗.