Fig. 2: In vitro cytokine induction assay.
a Human PBMCs from healthy donors were pre-cultured at high density (1 × 107 cells/ml) for 48 h to increase cell–cell contacts and immune scanning. The cells were then harvested and treated with anti-CD3 or anti-CD3 + anti-CD28 antibodies as positive controls, and CA-170 (10 nM to 1000 nM). Release of IL-2, IFN-γ, and TNF-α was measured in the culture supernatants at 48 h. Two donors were used in this study (one male and one female); there was no gender difference observed in the cytokine response in this study. Data presented is representative data from a male donor (BLoD Below Limit of Detection). b Cytokine levels determined in PBMCs from the same donor with similar experimental conditions but cultured at a normal density of 1 × 105 cells/well for 48 h for comparison. For a–b each bar in the figure denotes mean ± SD from the three replicates.
