Figure 5
Lipidated imidazoquinoline UM-3003 demonstrates robust potency and efficacy in inducing Th-polarizing cytokines from human newborn MoDCs. (A–B) Newborn MoDCs were stimulated for 24 h and supernatants were collected for multiplex assay. Cytokine responses (pg/ml) of 13 analytes (n = 5) were represented in radar plot (A) demonstrating neonatal MoDCs concurrently stimulated in the presence of 2% glycerin (vehicle control represented by thick black line) or 100 ng/ml of LPS (blue line) or 10 μM of UM-3003 (orange line). (B) Fold-change of 100 ng/ml of LPS (blue line) or 10 μM of UM-3003 (orange line) over vehicle control (thick black line) are shown. (C) TNF (n = 5), (D) IL-12p70 (n = 5) ELISA were performed by stimulated supernatants where lipidated compound demonstrating similar TNF induction ability compared with LPS, but slightly increased IL-12p70 production. In, (E–F), cord blood mononuclear cells (CBMCs) (n = 6) were stimulated in the presence of the polyclonal T cell activator α-CD3 (5 μg/ml) in combination or alone for 96 h. TLR7/8 agonist (CL075, 10 μM) and STING agonist (2′3′-cGAMP, 10 μM) in combination were used as a benchmark control. (E) Radar plot demonstrated total cytokine levels (pg/ml) in cell free supernatants by a high sensitivity T cell multiplex assay from six independent experiments. (F) Fold changes relative to vehicle control are shown. Statistical analysis was performed by nonparametric Kruskal–Wallis test corrected for multiple comparisons and statistical significance denoted as *p < 0.033, **p < 0.002.
