Basic–alimentary tractMyofibroblast Matrix Metalloproteinases Activate the Neutrophil Chemoattractant CXCL7 From Intestinal Epithelial Cells
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Cell Culture
Caco-2 human colon carcinoma cells (passages 30–35) were maintained in Dulbecco’s modified Eagle medium (Invitrogen, Paisley, Scotland) supplemented with 10% fetal calf serum (Invitrogen) as previously described.10 Caco-2 cells were seeded at a density of 3 × 105 on polycarbonate culture plate inserts (0.4 μm pore size; Millipore, Watford, England), which were placed in 6-well plates and incubated in 2 mL of apical medium and 2 mL of basolateral medium. Monolayer formation was followed by
MMP Activity Induces Caco-2 Cells to Attract Neutrophils
We first addressed whether MMPs could modulate the ability of intestinal epithelial cell supernatants to attract leukocytes. Caco-2 epithelial cells, a recognized model of the intestinal epithelium,10 were grown for 21–28 days on filters and then basolaterally exposed to 0.5–50 nmol/L of active recombinant human MMP-3 in the absence or presence of 1 ng/mL IL-1β. After 24 hours, the conditioned basolateral media were tested in chemotaxis assays (Figure 1A and B). MMP-3 induced a dose-dependent,
Discussion
This work identifies a novel, proinflammatory action of MMP activity in mucosal immune activation. Subepithelial myofibroblast MMPs are directly involved in local leukocyte recruitment by eliciting a functional chemokine response from the intestinal epithelium, mediated by the neutrophil chemokine CXCL7 (NAP-2).
CXCL7, not previously identified in the intestine, is the only known neutrophil chemokine that is produced as a chemotactically inactive precursor protein, in this case PBP.18 PBP (94
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Supported by grants from the Medical Research Council and the St. Bartholomew’s and the Royal London Charitable Foundation.