Mechanisms of activation of the RAW264.7 macrophage cell line by transfected mammalian DNA.
Bacterial DNA can stimulate the production of cytokines and nitric oxide (NO), while mammalian DNA can block these responses. If mammalian DNA is transfected into macrophages, however, it can stimulate NO production, without inducing IL-12. To define further this activity, signaling pathways induced by transfected calf thymus (CT) DNA were studied. Using RAW264.7 cells as a model, CT DNA in the transfection agent FuGENE 6 activated cells through the NF-kappaB and MAPKs pathways, similar to bacterial DNA and LPS. The role of these pathways was further investigated using specific inhibitors, with studies indicating that NO production is blocked by inhibitors of NF-kappaB and p38 but not other MAPKs. These data indicate that the immune activity of DNA is influenced by context or intracellular location and that, when transfected into cells, mammalian DNA can activate cells through signaling pathways similar to those of bacterial DNA.
Department of Medicine, Division of Rheumatology and Immunology, Duke University, Durham, NC, USA.
Weiwen
W
Jiang
Charles F
CF
Reich III
David S
DS
Pisetsky
D000818:Animals; D016207:Cytokines / Q000096:biosynthesis; D004247:DNA / Q000378:metabolism*; D007093:Imidazoles / Q000378:metabolism; D050356:Lipid Metabolism; D008055:Lipids; D008262:Macrophage Activation / Q000502:physiology*; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism; D011725:Pyridines / Q000378:metabolism; D014162:Transfection
D016207:Cytokines; C411955:FuGene; D007093:Imidazoles; D008055:Lipids; D016328:NF-kappa B; D011725:Pyridines; D009569:Nitric Oxide; D004247:DNA; D020928:Mitogen-Activated Protein Kinases; C093642:SB 203580