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PMID: 11110796 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Induction of nitric-oxide synthase and activation of NF-kappaB by interleukin-12 p40 in microglial cells.

The Journal of biological chemistry ·Vol. 276 ·No. 11 ·2001-03-16 ·Pages 7899-905

Pahan K, Sheikh FG, Liu X, Hilger S, McKinney M, Petro TM

Abstract

Interleukin-12 (IL-12) is composed of two different subunits, p40 and p35. Expression of p40 mRNA but not that of p35 mRNA in excessive amount in the central nervous system of patients with multiple sclerosis (MS) suggests that IL-12 p40 may have a role in the pathogenesis of the disease. However, the mode of action of p40 is completely unknown. Because nitric oxide produced from the induction of nitric-oxide synthase (iNOS) also plays a vital role in the pathophysiology of MS, the present study was undertaken to explore the role of p40 in the induction of NO production and the expression of iNOS in microglia. Both IL-12 and p40(2), the p40 homodimer, dose-dependently induced the production of NO in BV-2 microglial cells. This induction of NO production was accompanied by an induction of iNOS protein and mRNA. Induction of NO production by the expression of mouse p40 cDNA but not that of the mouse p35 cDNA suggests that the p40 but not the p35 subunit of IL-12 is involved in the expression of iNOS. In addition to BV-2 glial cells, p40(2) also induced the production of NO in mouse primary microglia and peritoneal macrophages. However, both IL-12 and p40(2) were unable to induce the production of NO in mouse primary astrocytes. Because activation of NF-kappaB is important for the expression of iNOS, we investigated the effect of p40(2) on the activation of NF-kappaB. Induction of the DNA binding as well as the transcriptional activity of NF-kappaB by p40(2) and inhibition of p40(2)-induced expression of iNOS by SN50, a cell-permeable peptide carrying the nuclear localization sequence of p50 NF-kappaB, but not by SN50M, a nonfunctional peptide mutant, suggests that p40(2) induces the expression of iNOS through the activation of NF-kappaB. This study delineates a novel role of IL-12 p40 in inducing the expression of iNOS in microglial cells, which may participate in the pathogenesis of neuroinflammatory diseases.

MeSH Terms
Animals Astrocytes/metabolism Cell Line Enzyme Induction/drug effects Interleukin-12/genetics,pharmacology Macrophages, Peritoneal/metabolism Mice Microglia/metabolism NF-kappa B/metabolism Nitric Oxide/biosynthesis Nitric Oxide Synthase/biosynthesis Nitric Oxide Synthase Type II Protein Subunits
Chemicals
NF-kappa B Protein Subunits Interleukin-12 Nitric Oxide Nitric Oxide Synthase Nitric Oxide Synthase Type II Nos2 protein, mouse
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Pahan K
Department of Oral Biology, University of Nebraska Medical Center, Lincoln, Nebraska 68583, USA. [email protected]
Sheikh F G
Liu X
Hilger S
McKinney M
Petro T M
References (56)
56 references, click to expand
  1. Induction of nitric oxide synthase in demyelinating regions of multiple sclerosis brains.
    Ann Neurol. 1994 Nov;36(5):778-86 PMID: 7526776
  2. Nitric oxide as a potential pathological mechanism in demyelination: its differential effects on primary glial cells in vitro.
    Neuroscience. 1994 Aug;61(3):575-85 PMID: 7969931
  3. Control of I kappa B-alpha proteolysis by site-specific, signal-induced phosphorylation.
    Science. 1995 Mar 10;267(5203):1485-8 PMID: 7878466
  4. Reactive nitrogen intermediates in human neuropathology: an overview.
    Dev Neurosci. 1994;16(3-4):152-61 PMID: 7535680
  5. Inhibition of nuclear translocation of transcription factor NF-kappa B by a synthetic peptide containing a cell membrane-permeable motif and nuclear localization sequence.
    J Biol Chem. 1995 Jun 16;270(24):14255-8 PMID: 7782278
  6. The IL-12 p40 homodimer as a specific antagonist of the IL-12 heterodimer.
    Immunol Today. 1995 Oct;16(10):500-1 PMID: 7576055
  7. Cytokine gene expression in islets and thyroids of BB rats. IFN-gamma and IL-12p40 mRNA increase with age in both diabetic and insulin-treated nondiabetic BB rats.
    J Immunol. 1996 Feb 1;156(3):1315-21 PMID: 8558012
  8. Nitric oxide: a neural messenger.
    Annu Rev Cell Dev Biol. 1995;11:417-40 PMID: 8689564
  9. Differential regulation by cytokines of human astrocyte nitric oxide production.
    Glia. 1995 Dec;15(4):491-4 PMID: 8926042
  10. A functional interleukin 12 receptor complex is composed of two beta-type cytokine receptor subunits.
    Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):14002-7 PMID: 8943050
  11. Regulation of inducible nitric oxide synthase messenger RNA expression and nitric oxide production by lipopolysaccharide in vivo: the roles of macrophages, endogenous IFN-gamma, and TNF receptor-1-mediated signaling.
    J Immunol. 1997 Jan 15;158(2):905-12 PMID: 8993010
  12. Inhibition of nitric oxide synthase for treatment of experimental autoimmune encephalomyelitis.
    J Immunol. 1997 Mar 15;158(6):2940-6 PMID: 9058833
  13. Increasing cAMP attenuates induction of inducible nitric-oxide synthase in rat primary astrocytes.
    J Biol Chem. 1997 Mar 21;272(12):7786-91 PMID: 9065441
  14. A cytokine-responsive IkappaB kinase that activates the transcription factor NF-kappaB.
    Nature. 1997 Aug 7;388(6642):548-54 PMID: 9252186
  15. IL-12 production by central nervous system microglia is inhibited by astrocytes.
    J Immunol. 1997 Aug 15;159(4):1604-12 PMID: 9257819
  16. IKK-1 and IKK-2: cytokine-activated IkappaB kinases essential for NF-kappaB activation.
    Science. 1997 Oct 31;278(5339):860-6 PMID: 9346484
  17. Catalysis by a multiprotein IkappaB kinase complex.
    Science. 1997 Oct 31;278(5339):818-9 PMID: 9381193
  18. Lovastatin and phenylacetate inhibit the induction of nitric oxide synthase and cytokines in rat primary astrocytes, microglia, and macrophages.
    J Clin Invest. 1997 Dec 1;100(11):2671-9 PMID: 9389730
  19. N-acetyl cysteine inhibits induction of no production by endotoxin or cytokine stimulated rat peritoneal macrophages, C6 glial cells and astrocytes.
    Free Radic Biol Med. 1998 Jan 1;24(1):39-48 PMID: 9436612
  20. Sphingomyelinase and ceramide stimulate the expression of inducible nitric-oxide synthase in rat primary astrocytes.
    J Biol Chem. 1998 Jan 30;273(5):2591-600 PMID: 9446561
  21. Nuclear factor-kappa B contributes to excitotoxin-induced apoptosis in rat striatum.
    Mol Pharmacol. 1998 Jan;53(1):33-42 PMID: 9443930
  22. Expression of IL-12 in CNS and lymphoid organs of mice with experimental allergic encephalitis.
    J Neuroimmunol. 1998 Feb;82(1):22-30 PMID: 9526842
  23. Production of interleukin-12 and expression of its receptors by murine microglia.
    Brain Res. 1998 Mar 16;787(1):139-42 PMID: 9518583
  24. Inhibitors of protein phosphatase 1 and 2A differentially regulate the expression of inducible nitric-oxide synthase in rat astrocytes and macrophages.
    J Biol Chem. 1998 May 15;273(20):12219-26 PMID: 9575170
  25. The interleukin-12/interleukin-12-receptor system: role in normal and pathologic immune responses.
    Annu Rev Immunol. 1998;16:495-521 PMID: 9597139
  26. Increased release of interleukin-12p40 in MS: association with intracerebral inflammation.
    Neurology. 1998 Sep;51(3):753-8 PMID: 9748022
  27. Inhibition of phosphatidylinositol 3-kinase induces nitric-oxide synthase in lipopolysaccharide- or cytokine-stimulated C6 glial cells.
    J Biol Chem. 1999 Mar 12;274(11):7528-36 PMID: 10066820
  28. Decreased interleukin-10 and increased interleukin-12p40 mRNA are associated with disease activity and characterize different disease stages in multiple sclerosis.
    Ann Neurol. 1999 Jun;45(6):695-703 PMID: 10360761
  29. Expression of a constitutively active form of phosphatidylinositol 3-kinase inhibits the induction of nitric oxide synthase in human astrocytes.
    FEBS Lett. 2000 Apr 28;472(2-3):203-7 PMID: 10788611
  30. Uric acid, a peroxynitrite scavenger, inhibits CNS inflammation, blood-CNS barrier permeability changes, and tissue damage in a mouse model of multiple sclerosis.
    FASEB J. 2000 Apr;14(5):691-8 PMID: 10744626
  31. Characterization of mouse interleukin-12 p40 homodimer binding to the interleukin-12 receptor subunits.
    Eur J Immunol. 1999 Jun;29(6):2007-13 PMID: 10382764
  32. Expression of a dominant-negative mutant of p21(ras) inhibits induction of nitric oxide synthase and activation of nuclear factor-kappaB in primary astrocytes.
    J Neurochem. 2000 Jun;74(6):2288-95 PMID: 10820188
  33. Murine macrophages stimulated with central and peripheral nervous system myelin or purified myelin proteins release inflammatory products.
    Neurosci Lett. 2000 Jun 30;287(3):171-4 PMID: 10863022
  34. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  35. Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue.
    J Cell Biol. 1980 Jun;85(3):890-902 PMID: 6248568
  36. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  37. Characterization of ameboid microglia isolated from developing mammalian brain.
    J Neurosci. 1986 Aug;6(8):2163-78 PMID: 3018187
  38. Differentiation of murine macrophages to express nonspecific cytotoxicity for tumor cells results in L-arginine-dependent inhibition of mitochondrial iron-sulfur enzymes in the macrophage effector cells.
    J Immunol. 1988 Apr 15;140(8):2829-38 PMID: 2451695
  39. Peroxynitrite oxidation of sulfhydryls. The cytotoxic potential of superoxide and nitric oxide.
    J Biol Chem. 1991 Mar 5;266(7):4244-50 PMID: 1847917
  40. Cloning of cDNA for natural killer cell stimulatory factor, a heterodimeric cytokine with multiple biologic effects on T and natural killer cells.
    J Immunol. 1991 May 1;146(9):3074-81 PMID: 1673147
  41. Nitric oxide mediates glutamate neurotoxicity in primary cortical cultures.
    Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6368-71 PMID: 1648740
  42. DNA deaminating ability and genotoxicity of nitric oxide and its progenitors.
    Science. 1991 Nov 15;254(5034):1001-3 PMID: 1948068
  43. Cloning and expression of murine IL-12.
    J Immunol. 1992 Jun 1;148(11):3433-40 PMID: 1350290
  44. Nitric oxide: a pathogenetic factor in autoimmunity.
    Immunol Today. 1992 May;13(5):157-60 PMID: 1379434
  45. Nitric oxide as a secretory product of mammalian cells.
    FASEB J. 1992 Sep;6(12):3051-64 PMID: 1381691
  46. I kappa B interacts with the nuclear localization sequences of the subunits of NF-kappa B: a mechanism for cytoplasmic retention.
    Genes Dev. 1992 Oct;6(10):1899-913 PMID: 1340770
  47. Induction of calcium-independent nitric oxide synthase activity in primary rat glial cultures.
    Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10945-9 PMID: 1279698
  48. NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway.
    Science. 1993 Mar 26;259(5103):1912-5 PMID: 8096091
  49. In vivo expression of inducible nitric oxide synthase in experimentally induced neurologic diseases.
    Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):3024-7 PMID: 7681993
  50. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages.
    Science. 1993 Apr 23;260(5107):547-9 PMID: 8097338
  51. Suppression of arthritis by an inhibitor of nitric oxide synthase.
    J Exp Med. 1993 Aug 1;178(2):749-54 PMID: 7688035
  52. Microglial cell cytotoxicity of oligodendrocytes is mediated through nitric oxide.
    J Immunol. 1993 Aug 15;151(4):2132-41 PMID: 8102159
  53. Lipopolysaccharide induces activation of CD14-associated protein tyrosine kinase p53/56lyn.
    J Biol Chem. 1993 Oct 5;268(28):20725-8 PMID: 7691802
  54. Induction of nitric oxide synthase in rat C6 glioma cells.
    J Neurochem. 1994 Jan;62(1):315-21 PMID: 7505314
  55. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase.
    J Biol Chem. 1994 Feb 18;269(7):4705-8 PMID: 7508926
  56. Evidence for increased nitric oxide production in multiple sclerosis.
    J Neurol Neurosurg Psychiatry. 1995 Jan;58(1):107 PMID: 7823050
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M008262200
Email
[email protected]
Published
2001-03-16
Epub
2000-00-07
Pages
7899-905
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2041874
Subset
IM
Grants
NINDS NIH HHS · R01 NS039940 · United States
NINDS NIH HHS · R01 NS039940-01A1 · United States
NINDS NIH HHS · NS39940 · United States
External Links
PubMed source DOI: 10.1074/jbc.M008262200 PMC Full Text View journal details
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