Submitted on October 31, 2008
Revised on January 30, 2009
Accepted on March 12, 2009
Differential expression and regulation of ADAM17 and TIMP3 in acute inflamed intestinal epithelia
Annabelle Cesaro1, Abakar Abakar-Mahamat2, Patrick Brest3, Sandra Lassalle4, Eric SELVA5, Jérôme Filippi6, Xavier Hébuterne2, Jean-Pierre Hugot7, Alain Doglio3, Franck Galland8, Philippe Naquet8, Valérie Vouret-Craviari9, Baharia Mograbi3, and Paul M. Hofman1*
1 INSERM/UNSA
2 INSERM\UNSA\Archet Hospital
3 INSERM\UNSA
4 INSERM\UNSA\Pasteur Hospital
5 Pasteur Hospital
6 Archet Hospital
7 INSERM\University of Paris 7
8 INSERM\CNRS
9 INSERM \ UNSA
* To whom correspondence should be addressed. E-mail: hofman.p{at}chu-nice.fr.
The acute phase of Crohn's disease (CD) is characterized by a large afflux of polymorphonuclear leukocytes (PMNL) into the mucosa and by the release of TNF-
. Conversion of inactive TNF- into an active form requires the cleavage of a transmembrane TNF- precursor by the TNF--converting enzyme (ADAM17), a protease mainly regulated by the tissue inhibitor of metalloproteinase 3 (TIMP3). The aim of the present study was to investigate in an in vitro model of PMNL transepithelial migration and in the intestinal mucosa of patients with CD the expression and regulation of ADAM 17 and TIMP3 in intestinal epithelial cells (IEC). ADAM17 and TIMP3 expression was analyzed by Western blotting, RT-PCR, confocal microscopy and immunohistochemistry using the T84 model, and digestive biopsies. ADAM17 expression in IEC was increased at a post-transcriptional level during the early phase (from 2 to 4 h) of PMNL transepithelial migration while TIMP3 was only increased 24 h later. TNF- induced an early upregulation of ADAM17 in T84 cells, whereas PMNL adhesion, H2O2 or epithelial tight junction opening, alone, did not affect the amount of ADAM17. Immunohistochemistry of intestinal biopsies revealed that strong expression of ADAM17 was associated with a high activity of CD. In contrast, TIMP3 was very poorly expressed in these biopsies. ADAM17 and TIMP3 profiling did not correlated with the NOD2/CARD15 status. The ADAM17 activity was higher both in the early phase of PMNL transepithelial migration and in active CD. These results showed early post-transcriptional-upregulation of ADAM17 in IEC linked to PMNL transepithelial migration and a high activity of CD.
