Evaluating classic (TLR4) and novel (dopaminergic) mediated inflammatory responses in human macrophages across distinct culture and serum supplementation conditions 3376

Description Classic toll-like receptor activation on macrophages drives inflammation, and our data indicate that the neurotransmitter dopamine can also drive inflammation in these cells. Our data in human monocyte derived macrophages (MDM) show that dopamine promotes an inflammatory phenotype. We also show in vitro culture environment can impact MDM function. We hypothesized that dopamine receptor (DR) expression patterns impact MDM inflammatory response, and that serum supplementation in vitro can alter this. We cultured MDM in DMEM or RPMI Medium +/- FBS or Macrophage Serum Free Media (M-SFM). MDM were untreated or stimulated with dopamine or LPS and assessed for changes in DR expression (RT-qPCR), cytokine production (AlphaLISA), transcriptional profile (RNA-seq), NF-kB activity, and morphology (High Content Imaging). Dopamine increased IL-6 secretion which correlated with the expression pattern of specific DR transcripts. Serum influenced MDM activation, with FBS supplementation and culture in M-SFM significantly altered the baseline transcriptome. M-SFM culture also significantly suppressed the NF-kB response to LPS while the impact of FBS supplementation on pharmacodynamics was related to the base media. Culture media and serum supplementation also drove morphological differences (area, length-to-width, perimeter-to-area ratio). These studies show dopamine drives inflammation in MDM and that in vitro microenvironment is an active component of experimentation. Funding Sources NIH DA057337 (PJG), NIH DA058051 (PJG), T32-MH079785 (supporting BC), F30AI179472 (BC), P30-MH092177 (WD), and the Department of Pharmacology and Physiology at Drexel University College of Medicine.