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Published ahead of print on June 25, 2004, doi:10.1165/rcmb.2004-0099OC

Am. J. Respir. Cell Mol. Biol., Volume 31, Number 4, October 2004, 413-422

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Submitted on March 23, 2004
Revised on June 25, 2004

Eosinophils adhere to VCAM-1 via podosomes

Mats W Johansson1*, Ming H Lye1, Steven R Barthel2, Allison K Duffy1, Douglas S Annis1, and Deane F Mosher2

1 Department of Medicine, University of Wisconsin, Madison, WI, USA, 2 Department of Medicine, University of Wisconsin, Madison, WI, USA; Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI, USA

* To whom correspondence should be addressed. E-mail: mwj{at}medicine.wisc.edu.

Vascular cell adhesion molecule-1 (VCAM-1) supports specific eosinophil adhesion via {alpha}4{beta}1 integrin. We tested the hypothesis that adhesive contacts formed by eosinophils on VCAM-1 are different from focal adhesions formed by adherent fibroblasts. Eosinophils adherent on VCAM-1 formed punctate adhesions that fit the criteria for podosomes, highly dynamic structures found in adherent transformed fibroblasts, osteoclasts, and macrophages. The structures contained {beta}1 integrin subunit, phosphotyrosine-containing proteins, punctate filamentous actin, and gelsolin, a podosome marker. In contrast, non- transformed fibroblasts on VCAM-1 formed peripheral focal adhesions that were positive for {alpha}4, {beta}1, phosphotyrosine, vinculin, talin, and paxillin; negative for gelsolin; and associated with microfilaments. PMA or tumor necrosis factor-{alpha} and interleukin-5 stimulated podosome formation in adherent eosinophils. Because podosomes in tumor cells are associated with extracellular matrix degradation, we analyzed the VCAM-1 layer. VCAM-1 was lost under adherent eosinophils but not under adherent fibroblasts. This loss was inhibited by the metalloproteinase inhibitor ortho-phenanthroline and correlated with expression and podosome localization of a membrane-tethered metalloproteinase, ADAM8. Podosome-mediated VCAM-1 clearance may be a mechanism to regulate eosinophil arrest and extravasation in allergic conditions such as asthma.




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