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and TNF-
in Mouse Macrophages1
,¶,§,*,
*
Division of Basic Sciences, Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206;
Department of Biochemistry and Molecular Genetics,
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-
and a number of
stimuli, including TNF-
. Recent work has shown that TNF-
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- and TNF-. All three kinases were activated during
costimulation with IFN- and TNF-. 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- and TNF-. 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-
receptor CD120a (p55) in the presence of IFN-.
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