Cellular activation by plasmid DNA in various macrophages in primary culture.

Macrophages are an important group of cells responsible for the inflammatory response to unmethylated CpG dinucleotide (CpG motif) in plasmid DNA (pDNA) via Toll-like receptor 9 (TLR9). This finding is primarily based on in vitro studies. Previous in vivo studies also have suggested that tissue macrophages are involved in inflammatory cytokine release in the circulation following intravenous administration of pDNA to mice. However, the relationship between the in vitro and in vivo studies has not been sufficiently clarified. To gain insight into which types of cells are responsible for the production of cytokines upon interaction with pDNA, peritoneal macrophages, splenic macrophages, hepatic nonparenchymal cells (NPCs) including Kupffer cells and mesangial cells were isolated from mice. All types of primary cultured cells, except for mesangial cells, express TLR9 at varying levels. Splenic macrophages and hepatic NPCs were activated to produce tumor necrosis factor-alpha (TNF-alpha) by naked pDNA, whereas peritoneal macrophages and mesangial cells were not. pDNA complexed with N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethyl-ammonium chloride/cholesterol liposome induced TNF-alpha in the splenic macrophages but not in the other cell types. These results indicate that splenic macrophages and hepatic NPCs are closely involved in TNF-alpha production in response to pDNA. (c) 2008 Wiley-Liss, Inc. and the American Pharmacists Association