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(Journal of Leukocyte Biology. 2000;68:529-537.)
© 2000 by Society for Leukocyte Biology

Sequential migration of neutrophils across monolayers of endothelial and epithelial cells

Frederik P. J. Mul*, Astrid E. M. Zuurbier*, Hans Janssen{dagger}, Jero Calafat{dagger}, Sandra van Wetering{ddagger}, Pieter S. Hiemstra{ddagger}, Dirk Roos* and Peter L. Hordijk*

* Central Laboratory of the Netherlands Blood Transfusion Service and Laboratory for Experimental and Clinical Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands;
{dagger} Division of Cell Biology, Netherlands Cancer Institute, Amsterdam, The Netherlands; and
{ddagger} Department of Pulmonology, Leiden University Medical Center, Leiden, The Netherlands

Correspondence: Peter L. Hordijk, Ph.D., Central Laboratory of the Netherlands Blood Transfusion Service, Plesmanlaan 125, 1066 CX Amsterdam, The Netherlands. E-mail: P_Hordijk{at}CLB.NL

In the course of granulocyte-dominated lung inflammation, granulocytes migrate across the endothelium and epithelium of the lung and cause severe tissue damage. To study this process in more detail, we developed a bilayer transmigration model composed of primary human endothelial and lung epithelial cells, simultaneously cultured on opposite sides of Transwell filters. Electron microscopical analysis showed that the morphology of the cells and the expression of junctional proteins remained unaltered and that matrix components were deposited onto the filter. Intriguingly, neutrophil migration was more efficient across the bilayers than across single epithelial monolayers and did not differ from migration across single endothelial monolayers. Coculture experiments showed that endothelial cells stimulated epithelial cells to release IL-6 and that epithelial cells enhanced release of IL-8 from endothelial cells. Together these data reveal bidirectional signaling and enhanced neutrophil migration in a transmigration model of primary human epithelial and endothelial cells.

Key Words: human • cytokines • transmigration • IL-1ß, IL-6, IL-8




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