DNA and its cationic lipid complexes induce CpG motif-dependent activation of murine dendritic cells.
Unmethylated CpG motifs in bacterial DNA, but not in vertebrate DNA, are known to trigger an inflammatory response of antigen-presenting cells (APC). In this study, we investigated the cytokine release from murine dendritic cells (DC) by the addition of various types of DNA in the free or complexed form with cationic lipids. Naked plasmid DNA and Escherichia coli DNA with immunostimulatory unmethylated CpG motifs induced pro-inflammatory cytokine secretion from granulocyte-macrophage colony-stimulating factor (GM-CSF)-cultured bone marrow-derived DC and the DC cell-line, DC2.4 cells, though vertebrate calf thymus DNA (CT DNA) with less CpG motifs did not. These characteristics differed from mouse peritoneal resident macrophages that do not respond to any naked DNA. The amount of cytokines released from the DC was significantly increased by complex formation with cationic lipids when CpG-motif positive DNAs were used. Unlike murine macrophages or Flt-3 L cultured DC, GM-CSF DC did not release inflammatory cytokines in response to the addition of CT DNA/cationic lipid complex, suggesting that the activation is completely dependent on CpG motifs. Taken together, the results of the present study demonstrate that murine DC produce pro-inflammatory cytokines upon stimulation with CpG-containing DNAs and the responses are enhanced by cationic lipids. These results also suggest that DC are the major cells that respond to naked CpG DNA in vivo, although both DC and macrophages will release inflammatory cytokines after the administration of a DNA/cationic lipid complex.
Yoshinaga
Takaharu
T
Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan.
Yasuda
Kei
K
Ogawa
Yoshiyuki
Y
Nishikawa
Makiya
M
Takakura
Yoshinobu
Y
D000818:Animals; D002478:Cells, Cultured; D018899:CpG Islands / Q000276:immunology*; D016207:Cytokines / Q000096:biosynthesis; D004247:DNA / Q000276:immunology*; D019175:DNA Methylation; D004269:DNA, Bacterial / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D018836:Inflammation Mediators / Q000378:metabolism; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D051217:Toll-Like Receptor 9 / Q000276:immunology
D016207:Cytokines; D004269:DNA, Bacterial; D018836:Inflammation Mediators; C495370:Tlr9 protein, mouse; D051217:Toll-Like Receptor 9; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D004247:DNA; C427203:calf thymus DNA