Poster Presentation: Modulation of Neuroinflammation by Interleukin-10 in Mouse Microglial Cells

Microglia are resident macrophages of the central nervous system that are important for maintaining brain homeostasis. Persistent activation of microglia is associated with neuroinflammation in neurodegenerative disorders. Interest in immortalized microglia cell lines as cell culture models to study their biology and role in neuroinflammation has increased in the past decade, with mouse microglial BV-2 cells emerging as an in vitro model for neuronal inflammation similar to primary mouse microglial cells.

We report here IL-10 pretreatment of BV-2 cells compared to primary microglia from CD-1 mice. A 45-plex Luminex immunoassay was used to measure IL-10 pretreatment attenuation of TNF-a, IL-6, CCL5/RANTES, and G-CSF secretion from cells treated with IFN-g and LPS. In addition, we demonstrate the effects of small molecule inhibitors MLN4924, tofacitinib (CP 690550), and Cardamonin on IL-10 pretreatment. MLN4924 inhibits the NFkB pathway and decreased secretion of multiple cytokines and chemokines, including TNF-a, IL-6, CCL2/MCP-1, CCL5, CCL3/MIP-1a, and CCL4/MIP-1b. Tofacitinib inhibits the Jak1/Tyk2-STAT3 pathway, decreasing STAT-3 TYR705 phosphorylation, but had little effect on IL-10 cytokine suppression. Interestingly, IL-10 pretreatment increased secretion of IL-1ra, an M2b microglia phenotype marker, and chemokines CCL2 and CCL3.

Multiplex immunoassays are a valuable tool for investigating the mechanisms of neuroinflammation in primary and immortalized cell lines.