Cyclooxygenase (COX) exists as two isoforms: COX-1, which is constitutively expressed in most cell types; and COX-2, which is inducible by lipopolysaccharide (LPS) and cytokines in a variety of cell types. Although previous studies have implicated two DNA binding proteins, interferon regulatory factor (IRF)-1 and IRF-2, in the regulation of LPS- and IFN-γ-induced COX-2, their effects in vivo and in vitro are not well-defined. Using real-time PCR, COX-2 gene expression in the livers and lungs of mice challenged in vivo and in macrophages stimulated with LPS in vitro was investigated in wild-type and in IRF-1 and IRF-2 knockout mice. In response to 35 mg/kg LPS, IRF-1-, but not IRF-2-deficient mice, exhibited much poorer induction of COX-2 gene expression in both the livers and lungs. In vitro, COX-2 mRNA levels were also poorly induced in IRF-1-deficient macrophages, while IRF-2-deficient macrophages exhibited higher levels than in normal macrophages. IRF-1 and IRF-2 were confirmed to activate and repress expression of the COX-2 promoter, respectively, in a transient transfection system and the role of specific DNA binding sites confirmed by site-specific mutagenesis. Collectively, these data provide evidence for an important role for IRF-1 in vivo and in vitro and for IRF-2 in vitro in the regulation of COX-2 expression by LPS.
References
1.
Guha M., Mackman N.LPS induction of gene expression in human monocytes. Cell Signal2001; 13: 85—94.
2.
Sweet MJ, Hume DAEndotoxin signal transduction in macrophages. J Leukoc Biol1996; 60: 8—26.
3.
Funk CDProstaglandins and leukotrienes: advances in eicosanoid biology. Science2001; 294: 1871—1875.
4.
Smith WL, Dewitt DLBiochemistry of prostaglandin endoperoxide H systhase-1 and synthase-2 and their differential susceptibility to nonsteroidal anti-inflammtory drugs. Semin Nephrol1995; 15: 179—194.
5.
Stack E., DuBois RNRegulation of cyclo-oxygenase-2. Best Pract ResClin Gastroenterol2001; 15: 787—800.
6.
Mamane Y., Heylbroeck C., Genin P. et al. Interferon regulatory factors: the next generation. Gene1999; 237: 1—14.
7.
Taniguchi T. , Ogasawara K., Takaoka A., Tanaka N.IRF family of transcription factors as regulators of host defense. Annu Rev Immunol2001; 19: 623—655.
8.
Manthey CL, Perera PY, Salkowshi CA, Vogel SNTaxol provides a second signal for murine macrophage tumoricidal activity . J Immunol1994; 152: 825—831.
9.
Fujihara M., Ito N., Pace JL et al. Role of endogenous interferon-beta in lipopolysaccharide-triggered activation of the inducible nitric-oxide synthase gene in a mouse macrophage cell line, J774. J Biol Chem1994; 269: 12773—12778.
10.
Ohmori Y., Hamilton TARequirement for STAT1 in LPS-induced gene expression in macrophages. J Leukoc Biol2001; 69: 598—604.
11.
Lohoff M., Duncan GS, Ferrick D. et al. Deficiency in the transcription factor interferon regulatory factor (IRF)-2 leads to severely compromised development of natural killer and T helper type 1 cells. J Exp Med2000; 192: 325—335.
12.
Blaine SA, Wick M., Dessev C., Nemenoff RAInduction of cPLA2 in lung epithelial cells and non-small cell lung cancer is mediated by Sp1 and c-Jun. J Biol Chem2001; 276: 42737—42743.
13.
Mestre JR, Mackrell PJ, Rivadeneria DE et al. Redundancy in the signaling pathways and promoter elements regulating cyclooxygenase-2 gene expression in endotoxin-treated macrophage/monocytic cells. J Biol Chem2000; 276: 3977—3982.
14.
Subbaramaiah K., Lin DT, Hart JC, Dannenberg AJPeroxisome proliferator-activated receptor gamma ligands suppress the transcriptional activation of cyclooxygenase-2. Evidence for involvement of activator protein-1 and CREB-binding protein/p300 . J Biol Chem2001; 276: 12440—12448.
15.
Kirtikara K. , Raghow R., Laulederkind SJ et al. Transcriptional regulation of cyclooxygenase-2 in the human microvascular endothelial cell line, HMEC-1: control by the combinational reactions of AP2, NF-IL-6 and CRE elements. Mol Cell Biochem2000; 203: 41—51.
16.
Caivano M., Gorgoni B., Cohen P., Poli V.The induction of cyclooxygenase-2 mRNA in macrophages is biphasic and requires both C/EBP{beta} and C/EBP{delta} transcription factors. J Biol Chem2001; 276: 48693—48701.
17.
Tang Q., Chen W., Gonzales MS et al. Role of cyclic AMP responsive element in the UVB induction of cyclooxygenase-2 transcription in human keratinocytes. Oncogene2001; 20: 5164—5172.
18.
Blanco Jcg, Contursi C., Vogel SN et al. Interferon regulatory factor (IRF)-1 and IRF-2 regulate interferon γ-dependent cyclooxygenase 2 expression. J Exp Med2000 ; 191: 2131—2144.
19.
Kobayashi O. , Miwa H., Watanabe S. et al. Cyclooxygenase-2 downregulates inducible nitric oxide synthase in rat intestinal epithelial cells. Am J Physiol2001; 281: G688—G696.
20.
Barrios-Rodiles M., Chadee K.Novel regulation of cyclooxygenase-2 expression and prostaglandin E2 production by IFN-γ in human macrophages. J Immunol1998; 161: 2441—2448.
21.
Nogawa SF, Zhang F., Ross ME, Ladecola C.Cyclo-oxygenase-2 gene expression in neurons contributes to ischemic brain damage. J Neurosci1997; 17: 2746—2755.
22.
Bazan NGCOX-2 as a multifunctional neuronal modulator. Nat Med2001; 7: 414—415.
23.
Iadecola C. , Salkowski CA, Zhang F. et al. The transcription factor interferon regulatory factor 1 is expressed after cerebral ischemia and contributes to ischemic brain injury. J Exp Med1999; 189: 719—727.
24.
Perera PY, Mayadas TN, Takeuchi O. et al. CD11b/CD18 acts in concert with CD14 and Toll-like receptor (TLR) 4 to elicit full lipopolysaccharide and Taxol-inducible gene expression . J Immunol2001; 166: 574—581.
25.
Ogura S., Umegaki O., Genovese F. et al. Protective effect of tyrphostin AG-556 on shock induced by endotoxin or Gram-positive bacteria. Shock1999; 12: 105—110.
26.
Huang YC, Guh JH, Cheng ZJ et al. Inhibitory effect of DCDC on lipopolysaccharide-induced nitric oxide synthesis in RAW 264.7 cells. Life Sci2001; 68: 2435—2447.
27.
Liu SF, Ye X., Malik ABInhibition of NF-kappaB activation by pyrrolidine dithiocarbamate prevents in vivo expression of pro-inflammatory genes. Circulation1999; 100: 1330—1337.
28.
Futaki N., Takahashi S., Kitagawa T. et al. Selective inhibition of cyclooxygenase-2 by NS-398 in endotoxin shock rats in vivo. Inflamm Res1997; 46: 496—502.
29.
McIntire FC , Sievert HW, Barlow GH et al. Chemical, physical, biological properties of lipopolysaccharide from Escherichia coli K-235. Biochemistry1967; 6: 2363—2372.
30.
Wang IM, Blanco JC, Tsai SY et al. Interferon regulatory factors and TFIIB co-operatively regulate interferon-responsive promoter activity in vivo and in vitro. Mol Cell Biol1996; 16: 6313—6324.
31.
Masumi A., Wang IM, Lefebvre B. et al. The histone acetylase PCAF is a phorbol-ester-inducible co-activator of the IRF family that confers enhanced interferon responsiveness. Mol Cell Biol1999; 19: 1810—1820.
32.
Kraemer SA, Arthur KA, Denison MS et al. Regulation of prostaglandin endoperoxide H synthase-2 expression by 2,3,7,8-tetrachlorodibenzop-dioxin. Arch Biochem Biophys1996; 330: 319—328.
33.
Salkowski CA, Thomas KE, Cody MJ, Vogel SNImpaired IFNgamma production in IFN regulatory factor-1 knockoutmice during endotoxemia is secondary to a loss of both IL-12 and IL-12 receptor expression . J Immunol2000; 165: 3970—3977.
34.
Wieland CW, Siegmund B., Senaldi G. et al. Pulmonary inflammation induced by Pseudomonas aeruginosa lipopolysaccharide, phospholipase C, and exotoxin A: role of interferon regulatory factor 1. Infect Immun2002; 70: 1352—1358.
35.
Toshchakov V., Jones BW, Perera PY et al. TLR4, but not TLR2, mediates IFN-β-induced STAT1 α / β -dependent gene expression in macrophages. Nat Immunol2002; 3: 392—398.
36.
Okamoto T., Hino O.Expression of cyclooxygenase-1 and —2 mRNA in rat tissues: tissue-specific difference in the expression of the basal level of mRNA. Int J Mol Med2000; 6: 455—457.
37.
Soslow RA, Dannenberg AJ, Rush D. et al. COX-2 is expressed in human pulmonary, colonic, and mammary tumors . Cancer2000; 89: 2637—2645.
38.
Shuhji S., Yoshiko H., Toshihiko K. et al. The liver as a crucial organ in the first line of host defense: the roles of Kupffer cells, natural killer (NK) cells and NK1.1 Ag+ T cells in T helper 1 immune responses. Immunol Rev2000; 174: 35—46.
39.
Whitsett JAIntrinsic and innate defenses in the lung: intersection of pathways regulating lung morphogenesis, host defense, and repair. J Clin Invest2002; 109: 565—569.
40.
Saitoh H., Masuda T., Zhang XY et al. Effect of antisense oligonucleotides to nuclear factor-kappaB on the survival of LPS-induced ARDS in mouse. Exp Lung Res2002; 28: 219—231.
