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B in Cytokine Production Induced from Human Airway Epithelial Cells by Rhinovirus Infection1
Divisions of
*
Clinical Immunology and
Pulmonary and Critical Care Medicine, Department of Medicine, and
Department of Otolaryngology-Head and Neck Surgery, Johns Hopkins University School of Medicine, Baltimore, MD 21224
Infection of human epithelial cells with human rhinovirus (HRV)-16
induces rapid production of several proinflammatory cytokines,
including IL-8, IL-6, and GM-CSF. We evaluated the role of NF-
B in
HRV-16-induced IL-8 and IL-6 production by EMSA using oligonucleotides
corresponding to the binding sites for NF-B in the IL-6 and IL-8
gene promoters. Consistent with the rapid induction of mRNA for IL-8
and IL-6, maximal NF-B binding to both oligonucleotides was detected
at 30 min after infection. NF-B complexes contained p65 and p50, but
not c-Rel. The IL-8 oligonucleotide bound recombinant p50 with only
about one-tenth the efficiency of the IL-6 oligonucleotide, even though
epithelial cells produced more IL-8 protein than IL-6. Neither the
potent glucocorticoid, budesonide (10-7 M), nor a NO donor
inhibited NF-B binding to either cytokine promoter or induction of
mRNA for either IL-8 or IL-6. Sulfasalazine and calpain inhibitor I,
inhibitors of NF-B activation, blocked HRV-16-induced formation of
NF-B complexes with oligonucleotides from both cytokines, but did
not inhibit mRNA induction for either cytokine. By contrast,
sulfasalazine clearly inhibited HRV-16 induction of mRNA for GM-CSF in
the same cells. Thus, HRV-16 induces epithelial expression of IL-8 and
IL-6 by an NF-B-independent pathway, whereas induction of GM-CSF is
at least partially dependent upon NF-B
activation.
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