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The Journal of Immunology, 1999, 162: 415-422.
Copyright © 1999 by The American Association of Immunologists

Evaluation of the Role of Mitogen-Activated Protein Kinases in the Expression of Inducible Nitric Oxide Synthase by IFN-{gamma} and TNF-{alpha} in Mouse Macrophages1

Edward D. Chan{ddagger}, Brent W. Winston||, Soo-Taek Uh{ddagger}, Murry W. Wynes, David M. Rose§ and David W. H. Riches2,{ddagger},*,{dagger}

* Division of Basic Sciences, Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206; {dagger} Department of Biochemistry and Molecular Genetics, {ddagger} Division of Pulmonary Sciences and Critical Care Medicine, Department of Medicine, § Department of Pharmacology, and Department of Immunology, University of Colorado Health Sciences Center, Denver, CO 80262; and || Faculty of Medicine, Division of Critical Care Medicine, University of Calgary, Calgary, Alberta, Canada

The expression of inducible nitric oxide synthase (iNOS) by macrophages is stimulated by coexposure to IFN-{gamma} and a number of stimuli, including TNF-{alpha}. Recent work has shown that TNF-{alpha} activates members of the mitogen-activated protein kinase family that subsequently trans-activate transcription factors implicated in the regulation of iNOS expression. The objective of this study was to systematically evaluate the role of: 1) p42mapk/erk2, 2) p46 c-Jun NH2-terminal kinase/stress-activated protein kinase (p46 JNK/SAPK), and 3) p38mapk in the induction of iNOS expression during costimulation of mouse macrophages with IFN-{gamma} and TNF-{alpha}. All three kinases were activated during costimulation with IFN-{gamma} and TNF-{alpha}. However, specific antagonism of the p42mapk/erk2 and p38mapk with PD98059 and SKF86002, respectively, had no effect on the induction of iNOS expression. In contrast, blockade of all three kinases with N-acetylcysteine completely blocked the induction of iNOS expression. In addition, specific antagonism of the JNK/SAPK upstream kinases MEKK (mitogen-activated protein kinase/extracellular signal-regulated kinase kinase kinase) and MKK4 (mitogen-activated protein kinase kinase 4) with dominant inhibitory mutants blocked transcriptional activation of the iNOS promoter in response to costimulation with IFN-{gamma} and TNF-{alpha}. Collectively, these findings support the involvement of p46 JNK/SAPK and its upstream kinases in regulating the induction of iNOS following ligation of the TNF-{alpha} receptor CD120a (p55) in the presence of IFN-{gamma}.




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