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Published ahead of print on March 4, 2004, doi:10.1164/rccm.200306-855OC
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American Journal of Respiratory and Critical Care Medicine Vol 169. pp. 1110-1117, (2004)
© 2004 American Thoracic Society

Original Article

Contribution of Eotaxin-1 to Eosinophil Chemotactic Activity of Moderate and Severe Asthmatic Sputum

Gordon Dent, Chrystalleni Hadjicharalambous, Takahiro Yoshikawa, Rachel L. C. Handy, John Powell, Ian K. Anderson, Renaud Louis, Donna E. Davies and Ratko Djukanovic

Respiratory Cell and Molecular Biology, Division of Infection, Inflammation, and Repair, University of Southampton School of Medicine, Southampton; Cambridge Antibody Technology, Cambridge, United Kingdom; and Pneumology and Allergology, Faculty of Medicine, University of Liège, Liège, Belgium

Correspondence and requests for reprints should be addressed to Gordon Dent, Ph.D., School of Life Sciences, Huxley Building, Keele University, Keele, Staffordshire ST5 5BG, UK. E-mail: g.dent{at}hfac.keele.ac.uk

The CC chemokine eotaxin-1 (CCL11) is chemotactic for eosinophils, basophils, and type 2 helper T cells and may play a role in allergic inflammation. We investigated its contribution as an eosinophil chemoattractant in asthmatic airway secretions (sampled as induced sputum), which possess chemotactic activity for eosinophils and T cells. Sputum samples collected from healthy subjects and subjects with mild, stable-moderate, unstable-moderate, and severe asthma were processed with phosphate-buffered saline and assayed for eotaxin by ELISA and for eosinophil chemotactic activity by fluorescence-based chemotaxis assay. The contribution of eotaxin to chemotactic activity was studied by using a high-affinity neutralizing human anti-eotaxin antibody, CAT-213. Sputum eotaxin concentration was significantly raised in moderate and severe asthma (p < 0.05 versus healthy control subjects) but not in mild asthma. Chemotactic activity was significantly increased in all asthmatic groups relative to healthy subjects (p < 0.05) and was significantly inhibited by CAT-213 (100 nM) in subjects with moderate and severe asthma, with median inhibition of 52% (p < 0.05), 78% (p < 0.0001), and 86% (p < 0.0001), respectively, in samples representing stable-moderate, unstable-moderate, and severe asthma. Eotaxin contributed to the eosinophil chemotactic activity of sputum from subjects with more severe forms of asthma but not mild asthma, suggesting that its contribution is more important in more severe disease. This activity is inhibited significantly by CAT-213.

Key Words: antibodies • asthma • chemokines • chemotaxis




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