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Published ahead of print on December 4, 2003, doi:10.1165/rcmb.2003-0370OC

Am. J. Respir. Cell Mol. Biol., Volume 30, Number 6, June 2004, 761-770

A more recent version of this article appeared on June 1, 2004
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Submitted on October 16, 2003
Revised on December 3, 2003

MMP-9 DEFICIENT DENDRITIC CELLS HAVE IMPAIRED MIGRATION THROUGH TRACHEAL EPITHELIAL TIGHT JUNCTIONS

Hidenori Ichiyasu1, Joanne M McCormack1, Karin M McCarthy1, David Dombkowski1, Frederic I Preffer1, and Eveline E Schneeberger1*

1 Pathology, Massachusetts General Hospital, Charlestown, MA, USA

* To whom correspondence should be addressed. E-mail: eschneeeberger{at}partners.org.

When sampling inhaled antigens, dendritic cells (DC) must penetrate the tight junction (TJ) barrier while maintaining the TJ seal. In MMP-9 deficient mice, in vivo experiments suggest that migration of DC into air spaces is impaired. To examine the underlying mechanisms, we established a well-defined in vitro model using mouse tracheal epithelial cells and mouse bone marrow DC (BMDC). Transmigration was elicited with either MIP-1{alpha} or MIP-3{beta} in a time dependent manner. Control MMP-9+/+ BMDC cultured with GM-CSF for 7 days showed a 30-fold greater trans-epithelial migration towards MIP-3{beta} than MIP-1{alpha}, indicating a more mature DC phenotype. MMP-9-/- BMDC as well as MMP-9+/+ BMDC in the presence of the MMP inhibitor GM6001, while showing a similar preference for MIP-3{beta}, were markedly impaired in their ability to traverse the epithelium. Expression levels of CCR5 and CCR7, however, were similar in both MMP-9-/- and MMP-9+/+ BMDC. Expression of the integral TJ proteins, occludin and claudin-1, were examined in BMDC before and after transepithelial migration. Interestingly, occludin but not claudin-1 was degraded following transepithelial migration in both MMP-9-/- and control BMDC. In addition, there was a greater than a two-fold increase in claudin-1 expression in MMP-9-/- as compared to control BMDC. These observations indicate that occludin and claudin-1 are differentially regulated and suggest that the lack of MMP-9 may affect claudin-1 turnover.




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