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1 Physiology Program, Harvard School of Public Health, Boston, Massachusetts 02115; and 2 Pulmonary and Critical Care Division, Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104
Human airway smooth muscle (HASM) cells
express interleukin (IL)-13 and IL-4 receptors and respond to these
cytokines with signal transducer and activator of transcription-6 and
extracellular signal-regulated kinase (ERK) activation. The purpose of
this study was to determine whether IL-13 and/or IL-4 influence eotaxin release in HASM cells and whether the ERK mitogen-activated protein (MAP) kinase pathway is involved in these events. Eotaxin release into
HASM cell supernatants was assayed by ELISA, and eotaxin mRNA
expression was determined by Northern blot analysis. Pretreatment with
either IL-13 or IL-4 resulted in a concentration- and time-dependent release of eotaxin, although IL-4 was more effective. Eotaxin release
was approximately twice baseline after treatment with 50 ng/ml IL-13 or
IL-4 (P < 0.001). IL-13 and IL-4 also acted synergistically with tumor necrosis factor (TNF)-
to induce eotaxin release: TNF- alone (10 ng/ml for 24 h) resulted in an
approximately fourfold increase in eotaxin release, whereas TNF- in
combination with IL-13 or IL-4 resulted in 10- or 20-fold increases
(P < 0.05). Similar results were obtained for eotaxin
mRNA expression. Pretreatment with either U-0126 (10 µM) or PD-98059
(30 µM), both inhibitors of MAP/ERK kinase, the enzyme upstream of
ERK, inhibited IL-13- or IL-4-induced eotaxin release
(P < 0.05). U-0126 also inhibited IL-13, and TNF-
induced mRNA expression. Our results indicate that IL-13 and IL-4 cause
eotaxin release in HASM cells through a mechanism that, in part,
involves ERK activation and suggest that the smooth muscle may be an
important source of chemokines leading to eosinophil recruitment in asthma.
U-0126; PD-98059; tumor necrosis factor-; asthma; extracellular
regulated kinase; interleukin
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