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Blood, Vol. 93 No. 10 (May 15), 1999:
pp. 3241-3249
NF- B Activation Is Required for C5a-Induced Interleukin-8 Gene
Expression in Mononuclear Cells
Matthew H. Hsu,
Meiying Wang,
Darren D. Browning,
Naofumi Mukaida, and
Richard D. Ye
From the Department of Immunology, The Scripps Research Institute, La
Jolla, CA; and the Department of Molecular Pharmacology, Cancer
Research Institute, Kanazawa University, Kanazawa, Ishikawa, Japan.
C5a, a potent peptide chemoattractant, stimulates interleukin-8
(IL-8) secretion from peripheral blood mononuclear cells (PBMC). Experiments were conducted to understand the mechanisms for C5a-induced IL-8 production, which was 14-fold greater than that in unstimulated cells by 2 hours. IL-8 secretion was accompanied by accumulation of
IL-8 mRNA in the cytosol and by nuclear expression of a  B DNA
binding activity within 30 minutes. AP-1 but not NF-IL-6 DNA binding
activity was also detected in C5a-stimulated PBMC; however, its delayed
expression (maximal at 4 hours) suggested a less important role in the
rapid production of IL-8. The correlation between C5a-induced B
binding activity and IL-8 gene expression was examined in the RAW264.7
macrophage cells using reporter genes directed by the B sequence
from IB and IL-8 promoter regions. C5a-induced reporter gene
expression was abolished by introducing mutations into the B sites
and by coexpression of a dominant negative IB construct resistant
to agonist-induced phosphorylation. Pertussis toxin, which
ADP-ribosylates the Gi proteins known to couple to the C5a
receptor, produced minimal inhibition of C5a-induced IL-8 expression
and had little effect on C5a-induced calcium mobilization in RAW264.7
cells. These results suggest that NF-B activation is required for
C5a-induced IL-8 gene expression and that this response is mediated
primarily through a pertussis toxin-insensitive pathway.
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