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Am. J. Respir. Crit. Care Med., Volume 159, Number 6, June 1999, 1933-1942

Constitutive and Cytokine-Stimulated Expression of Eotaxin by Human Airway Smooth Muscle Cells

OMAR GHAFFAR, QUTAYBA HAMID, PAOLO M. RENZI, ZOULFIA ALLAKHVERDI, SOPHIE MOLET, JAMES C. HOGG, STEPHANIE A. SHORE, ANDREW D. LUSTER, and BOUCHAIB LAMKHIOUED

Meakins-Christie Laboratories, McGill University, Montreal, Quebec, Canada; CHUM Research Center, Montreal, Quebec, Canada; Pulmonary Research Laboratory, University of British Columbia, Vancouver, British Columbia, Canada; Harvard School of Public Health, Boston; and Infectious Disease Unit, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts

Airway eosinophilia is a prominent feature of asthma that is believed to be mediated in part through the expression of specific chemokines such as eotaxin, a potent eosinophil chemoattractant that is highly expressed by epithelial cells and inflammatory cells in asthmatic airways. Airway smooth muscle (ASM) has been identified as a potential source of cytokines and chemokines. The aim of the present study was to examine the capacity of human ASM to express eotaxin. We demonstrate that airway myocytes constitutively express eotaxin mRNA as detected by RT-PCR. Treatment of ASM for 24 h with different concentrations of TNF-alpha and IL-1beta alone or in combination enhanced the accumulation of eotaxin transcripts. Maximal mRNA expression of eotaxin was shown at 12 and 24 h following IL-1beta and TNF-alpha stimulation, respectively. The presence of immunoreactive eotaxin was demonstrated by immunocytochemistry, and constitutive and cytokine-stimulated release of eotaxin was confirmed in ASM culture supernatants by ELISA. Strong signals for eotaxin mRNA and immunoreactivity were observed in vivo in smooth muscle in asthmatic airways. In addition, chemotaxis assays demonstrated the presence of chemoattractant activity for eosinophils and PBMCs in ASM supernatants. The chemotactic responses of eosinophils were partly inhibited with antibodies directed against eotaxin or RANTES, and a combined blockade of both chemokines causes > 70% inhibition of eosinophil chemotaxis. The results of this study suggest that ASM may contribute to airway inflammation in asthma through the production and release of eotaxin.




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