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Up-Regulates IL-18 Gene Expression Via IFN Consensus Sequence-Binding Protein and Activator Protein-1 Elements in Macrophages1
Laboratory of Immunology, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon, Republic of Korea
Constitutive IL-18 expression is detected from many different
cells, including macrophages, keratinocytes, and osteoblasts. It has
been known that IL-18 gene expression is regulated by two different
promoters (p1 promoter and p2 promoter). When RAW 264.7 macrophages
were treated with IFN-
, IL-18 gene expression was increased in a
dose- and time-dependent manner. IFN- activated the inducible
promoter 1, but not the constitutive promoter 2. Mutagenesis studies
indicated that an IFN consensus sequence-binding protein (ICSBP)
binding site between -39 and -22 was critical for the IFN-
inducibility. EMSA using an ICSBP oligonucleotide probe showed that
IFN- treatment increased the formation of DNA-binding complex, which
was supershifted with anti-IFN regulatory factor-1 Ab and
anti-ICSBP Ab. Another element, an AP-1 site between -1120 and
-1083, was important. EMSA using an AP-1-specific oligonucleotide
demonstrated that IFN- or LPS treatment increased the AP-1-binding
activity. The addition of anti-c-Jun Ab or anti-c-Fos Ab to
IFN-- or LPS-treated nuclear extracts resulted in the reduction of
AP-1 complex or the formation of a supershifted complex. Taken
together, these results indicate that IFN- increased IL-18 gene
expression via ICSBP and AP-1 elements.
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