Example 3
CD14+ monocytes were isolated from human PBMCs using negative magnetic bead selection (Stemcell Technologies) on day 0. Cells were cultured with M-CSF to differentiate monocytes to macrophages. On day 6, cells were treated with LP44 (high affinity 5-HT7 agonist), BW 723C86 hydrochloride (5-HT2B agonist), MK 212 hydrochloride (5-HT2C agonist) (Sigma-Aldrich), at concentrations 1 μM and 10 μM overnight. Supernatant was collected and bead-based immunoassays (Biolegend) were performed to detect changes in cytokine secretion.
Across multiple human donors, secretion of the inflammatory cytokine, IL8, by macrophages was decreased after addition of each of the HTR2B, HTR2C, and HTR7 agonists, as show in Table 17 below.