Myofibroblast matrix metalloproteinases activate the neutrophil chemoattractant CXCL7 from intestinal epithelial cells.
OBJECTIVE
The up-regulation of matrix metalloproteinases (MMPs) in the inflamed gut has mainly been associated with mucosal degradation and ulceration. However, their in vitro capacity to specifically cleave inflammatory mediators indicates that MMPs may have a profound immunoregulatory impact. We hypothesized that MMPs proteolytically modify intestinal epithelial chemokine signaling.


METHODS
Interleukin-1beta-stimulated Caco-2 cells were exposed basolaterally to nanomolar concentrations of activated MMP-3 or cocultured with interleukin-1beta-stimulated, MMP-producing, colonic myofibroblasts (CCD-18co). The conditioned media were subjected to chemotaxis assays. In addition, epithelial cells from patients with colitis were examined by real-time polymerase chain reaction, immunoblotting, and immunohistochemistry.


RESULTS
MMP-3 dose-dependently induced the neutrophil (up to 5-fold) but not monocyte chemoattractant capacity of Caco-2 cells. A similar Caco-2 chemotactic response was obtained in the Caco-2/CCD-18co cocultures. The principal mediator of these protease-related effects was identified as the potent neutrophil chemokine CXCL7 (neutrophil activating peptide 2), a proteolytic cleavage product of chemotactically inert platelet basic protein (PBP), not previously identified in the intestine. Antibodies against CXCL7 inhibited the MMP-induced chemotactic response by 84%, and PBP mRNA and protein were detected in stimulated Caco-2 but not in CCD-18co cells. Furthermore, PBP transcript and protein levels were low in the mucosa and in isolated epithelial cells from patients with Crohn's disease and from normal intestine but increased up to 13-fold in patients with ulcerative colitis.


CONCLUSIONS
These findings identify a novel proinflammatory action of MMPs in inflammation and suggest that lamina propria myofibroblasts are required to achieve maximal intestinal epithelial immune activation.
Kruidenier
Laurens
L
Research Centre in Gastroenterology, Institute of Cell and Molecular Science, Queen Mary's School of Medicine and Dentistry, University of London, London, England. l.kruidenier@qmul.ac.uk
MacDonald
Thomas T
TT
Collins
Jane E
JE
Pender
Sylvia L F
SL
Sanderson
Ian R
IR
D018938:Caco-2 Cells; D002633:Chemotaxis; D003093:Colitis, Ulcerative / Q000503:physiopathology; D003424:Crohn Disease / Q000503:physiopathology; D005347:Fibroblasts / Q000502:physiology; D006801:Humans; D007249:Inflammation; D007413:Intestinal Mucosa / Q000166:cytology / Q000276:immunology; D007962:Leukocytes / Q000502:physiology; D019278:Matrix Metalloproteinase 3 / Q000502:physiology; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000502:physiology*; D015398:Signal Transduction; D015854:Up-Regulation; D001620:beta-Thromboglobulin / Q000502:physiology*
C061615:PPBP protein, human; D001620:beta-Thromboglobulin; D020782:Matrix Metalloproteinases; D019278:Matrix Metalloproteinase 3