The Journal of Experimental Medicine
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Published 10 December 2001. doi:10.1084/jem.194.12.1699
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© Rockefeller University Press, 0022-1007/2001/12/1699/ $5.00
The Journal of Experimental Medicine, Volume 194, Number 12, December 17, 2001 1699-1709

Original Article

Shear-dependent Eosinophil Transmigration on Interleukin 4–stimulated Endothelial Cells : A Role for Endothelium-associated Eotaxin-3



Susan L. Cuvelier1 and Kamala D. Patel1,2

1 Department of Biochemistry and Molecular Biology, Immunology Research Group, University of Calgary, Calgary, Alberta, Canada T2N 4N1
2 Department of Physiology and Biophysics, Immunology Research Group, University of Calgary, Calgary, Alberta, Canada T2N 4N1

Address correspondence to Kamala D. Patel, 3330 Hospital Dr. NW, Calgary, Alberta, Canada T2N 4N1. Phone: 403-220-2746; Fax: 403-283-1267; E-mail: kpatel{at}ucalgary.ca

Leukocyte infiltration into inflammatory sites is regulated by the expression of adhesion and activation proteins, yet the role of these proteins in shear-dependent transmigration is poorly understood. We examined eosinophil recruitment on cytokine-stimulated human umbilical vein endothelial cells (HUVECs) under laminar flow conditions. Eosinophils rapidly transmigrated on interleukin (IL)-4–, but not TNF-stimulated HUVECs. Transmigration was shear dependent, with up to 90% of eosinophils transmigrating in the presence of shear and less than 25% of cells transmigrating under static conditions. Eosinophils express CC chemokine receptor CCR3 and are responsive to various CC chemokines. The effects of chemokines are mediated primarily through G{alpha}i, which is pertussis toxin sensitive. Greater than 65% of shear-dependent eosinophil transmigration on IL-4–stimulated HUVECs was blocked by either pertussis toxin or by an anti-CCR3 monoclonal antibody. Using reverse transcription polymerase chain reaction (RT-PCR) and Western blots, we found that IL-4–stimulated HUVECs produce both mRNA and protein for eotaxin-3. Eotaxin-3 was both released by HUVECs and expressed on the endothelial cell surface. Pretreatment of HUVECs with an anti–eotaxin-3 antibody blocked eosinophil transmigration to the same extent as an anti-CCR3 antibody. These results indicate that IL-4–stimulated HUVECs support shear-dependent eosinophil transmigration by upregulating eotaxin-3, and that surface association is critical for the role of eotaxin-3 in transmigration.

Key Words: chemokines • cell adhesion • cytokines • trafficking • leukocytes


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