Abstract
Oligodeoxynucleotides (ODN) containing CpG motifs activate RAW 264.7 mouse macrophages and RPMI 8226 human myeloma cells to produce IL-12 p40. Using deletion and site-directed mutagenesis, the nuclear factor (NF)-kappaB half-site and the CCAAT/enhancer binding protein (C/EBP) recognition site were identified as potent cis-acting elements in CpG ODN-mediated IL-12 p40 promoter activation. Several NF-kappaB/Rel proteins competed for binding to the NF-kappaB half-site. The p65/c-Rel and p65/p50 heterodimer occupied this site shortly after CpG ODN administration (0.5-2 h), while the p50/c-Rel heterodimer dominated binding in the late stage (8-12 h). The induction of p50/c-Rel heterodimer was associated with a significant expression of IL-12 p40 mRNA. C/EBPbeta also contributed to CpG ODN-mediated IL-12 p40 promoter activation.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Adjuvants, Immunologic / pharmacology*
-
Animals
-
Binding Sites
-
CCAAT-Enhancer-Binding Proteins
-
Cell Line
-
DNA-Binding Proteins / genetics
-
DNA-Binding Proteins / metabolism
-
Dimerization
-
Gene Expression Regulation / drug effects*
-
Humans
-
Interleukin-12 / genetics*
-
Macrophages / cytology
-
Macrophages / drug effects
-
Macrophages / immunology
-
Mice
-
NF-kappa B / genetics
-
NF-kappa B / metabolism
-
NF-kappa B p50 Subunit
-
Nuclear Proteins / genetics
-
Nuclear Proteins / metabolism
-
Oligodeoxyribonucleotides*
-
Oligonucleotides / pharmacology*
-
Promoter Regions, Genetic
-
Proto-Oncogene Proteins c-rel / genetics
-
Proto-Oncogene Proteins c-rel / metabolism
-
Thionucleotides / pharmacology*
-
Transcription Factor RelA
-
Transcription, Genetic / drug effects
-
Tumor Cells, Cultured
Substances
-
Adjuvants, Immunologic
-
CCAAT-Enhancer-Binding Proteins
-
CPG-oligonucleotide
-
DNA-Binding Proteins
-
NF-kappa B
-
NF-kappa B p50 Subunit
-
Nuclear Proteins
-
Oligodeoxyribonucleotides
-
Oligonucleotides
-
Proto-Oncogene Proteins c-rel
-
Thionucleotides
-
Transcription Factor RelA
-
Interleukin-12