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Blood, Vol. 94 No. 6 (September 15), 1999:
pp. 1878-1889
Nuclear Factor- B-Dependent Induction of Interleukin-8
Gene Expression by Tumor Necrosis Factor  : Evidence for an
Antioxidant Sensitive Activating Pathway Distinct From Nuclear
Translocation
Spiros Vlahopoulos,
Istvan Boldogh,
Antonella Casola, and
Allan R. Brasier
From the Departments of Internal Medicine, Microbiology & Immunology,
the Sealy Center for Molecular Sciences, and the Department of
Pediatrics, University of Texas Medical Branch, Galveston,
TX.
Tumor necrosis factor  (TNF) is a pluripotent activator of
inflammation by inducing a proinflammatory cytokine cascade. This
phenomenon is mediated, in part, through inducible expression of the
CXC chemokine, interleukin-8 (IL-8). In this study, we investigate the
role of TNF-inducible reactive oxygen species (ROS) in IL-8
expression by "monocyte-like" U937 histiocytic lymphoma cells.
TNF is a rapid activator of IL-8 gene expression by U937, producing
a 50-fold induction of mRNA within 1 hour of treatment. In gene
transfection assays, the effect of TNF requires the presence of an
inducible nuclear factor- B (NF-B) (Rel A) binding site in the
IL-8 promoter. TNF treatment induces a rapid translocation of the 65 kD transcriptional activator NF-B subunit, Rel A, whose binding in the nucleus occurs before changes in intracellular ROS.
Pretreatment (or up to 15 minutes posttreatment) relative to TNF
with the antioxidant dimethyl sulfoxide (DMSO) (2% [vol/vol]) blocks
80% of NF-B-dependent transcription. Surprisingly, however, DMSO
has no effect on inducible Rel A binding. Similar selective effects on
NF-B transcription are seen with the unrelated antioxidants, N-acetylcysteine (NAC) and vitamin C. These data indicate that TNF
induces a delayed ROS-dependent signalling pathway that is required for
NF-B transcriptional activation and is separable from that required
for its nuclear translocation. Further definition of this pathway will
yield new insights into inflammation initiated by TNF signalling.
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