Recruitment of eosinophils into the lung tissue is a critical event in allergic inflammatory reactions. Extravasation of eosinophils from the bloodstream is a highly dynamic multistep process that involves capture, rolling, activation, firm adhesion, and transendothelial and subendothelial migration of the cells. It is assumed that the rate-limiting step in this cascade is the capture and firm adhesion of cells to the endothelium. As such it is most critical to study endothelial–leukocyte interaction using assays which are capable of mimicking physiological flow conditions. Previously, various parallel flow chamber setups had been used for studying leukocyte adhesion to endothelial cells. Here we describe a highly reproducible technique for investigating eosinophil adhesion to endothelial cell layer or adhesion molecule/extracellular matrix protein coating in biochips by using a semiautomated microfluidic platform and live-cell imaging. In detail, we show eosinophil adhesion to endothelial cells activated by tumour necrosis factor (TNF) alpha, and adhesion to fibronectin of eosinophils stimulated by prostaglandin (PG) D2.
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This work was supported by the Austrian Science Fund FWF (Grant P25531 to V.K., P22521 to A.H.) and the Jubiläumsfonds of the Austrian National Bank (14263 to A.H.). M.P. was funded by the Ph.D. Program DK-MOLIN (W1241). The authors would like to thank Wolfgang Platzer and Iris Red for professional technical assistance.
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