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American Journal of Respiratory Cell and Molecular Biology. Vol. 28, pp. 713-721, 2003
© 2003 American Thoracic Society
DOI: 10.1165/rcmb.4771

Priming of Eosinophil Migration Across Lung Epithelial Cell Monolayers and Upregulation of CD11b/CD18 Are Elicited by Extracellular Ca2+

Lixin Liu, Lena Håkansson, Peter Ridefelt, Rodolfo C. Garcia and Per Venge

Department of Medical Sciences, Clinical Chemistry, Uppsala University, Uppsala, Sweden; and Leukocyte Biology Unit, International Center for Genetic Engineering and Biotechnology, Trieste, Italy

Address correspondence to: Dr. Lixin Liu, Immunology Research Group, HSC-1876, Health Sciences Centre, University of Calgary, 3330 Hospital Drive N.W., Calgary, AB, T2N 4N1 Canada. E-mail: lixliu{at}ucalgary.ca

In patients with asthma, eosinophils are primed and massively infiltrate lung tissues and migrate across epithelia into airways. Using blocking monoclonal antibodies, we found that eosinophil transmigration across a lung epithelial cell monolayer depended on the functions of {alpha}Mß2 integrin CD11b/CD18. To study the role of Ca2+ in eosinophil priming and transepithelial migration, we treated eosinophils with eotaxin or thapsigargin (TG), reagents that increase cytoplasmic free Ca2+ concentrations by receptor- or nonreceptor-mediated mechanisms, respectively. Pretreatment of eosinophils with TG enhanced CD11b/CD18-dependent transmigration across lung epithelium. Within minutes, TG time- and dose-dependently upregulated the expression of CD11b/CD18 but did not upregulate the expression of {alpha}L (CD11a) or ß1 (CD29) integrin. The upregulation of CD11b/CD18 expression by eotaxin or TG was prevented when Ca2+ entry was blocked. The priming of eosinophil transmigration by TG was also abrogated by the blockade of Ca2+ entry. Our results indicate that induction of Ca2+ entry by the depletion of Ca2+ from intracellular stores upregulates CD11b/CD18 expression on eosinophils and primes eosinophil transmigration across lung epithelium. Both responses are therefore elicited by extracellular Ca2+. We suggest that, as an important priming signal for human eosinophil functional responses, store-operated Ca2+ entry may be one of the underlying mechanisms of eosinophilic inflammation in asthma.

Abbreviations: bronchoalveolar lavage fluid, BALF • bovine serum albumin, BSA • Central Laboratory of the Netherlands Blood Transfusion Service, CLB • complement fragment 5a, C5a • cytoplasmic free Ca2+ concentration, [Ca2+]i • eosinophil cationic protein, ECP • fluorescein isothiocyanate, FITC • fura-2/acetoxymethyl ester, fura-2/AM • interleukin, IL • monoclonal antibodies, mAbs • mean fluorescence intensity, MFI • platelet-activating factor, PAF • phosphate-buffered saline, PBS • recombinant human, rh • thapsigargin, TG




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