Matrix metalloproteinase processing of monocyte chemoattractant proteins generates CC chemokine receptor antagonists with anti-inflammatory properties in vivo

Abstract

Monocyte chemoattractant protein (MCP)–3 is inactivated upon cleavage by the matrix metalloproteinase (MMP) gelatinase A (MMP-2). We investigated the susceptibility to proteolytic processing of the 4 human MCPs by 8 recombinant MMPs to determine whether MCP-3 is an isolated example or represents a general susceptibility of chemokines to proteolytic inactivation by these important inflammatory proteases. In addition to MMP-2, MCP-3 is efficiently cleaved by membrane type 1 (MT1)–MMP, the cellular activator of MMP-2, and by collagenase-1 and collagenase-3 (MMP-1, MMP-13) and stromelysin-1 (MMP-3). Specificity was shown by absence of cleavage by matrilysin (MMP-7) and the leukocytic MMPs neutrophil collagenase (MMP-8) and gelatinase B (MMP-9). The closely related chemokines MCP-1, MCP-2, and MCP-4 were not cleaved by MMP-2 or MT1-MMP, but were cleaved by MMP-1 and MMP-3 with varying efficiency. MCPs were typically cleaved between residues 4 and 5, but MCP-4 was further processed at Val7-Pro8. Synthetic MCP analogs corresponding to the MMP-cleaved forms bound CC chemokine receptor (CCR)–2 and CCR-3, but lacked chemoattractant activity in pre-B cells transfected with CCR-2 and CCR-3 or in THP-1 monocytic cells, a transformed leukemic cell line. Moreover, the truncated products of MCP-2 and MCP-4, like MCP-3, were potent antagonists of their cognate CC chemokine receptors in transwell cell migration assays in vitro. When they were injected 24 hours after the initiation of carrageenan-induced inflammation in rat paws, their in vivo antagonist activities were revealed by a greater than 66% reduction in inflammatory edema progression after 12 hours. We propose that MMPs have an important role in modulating inflammatory and immune responses by processing chemokines in wound healing and in disease.

MMPs and ADAMs in neurological infectious diseases and multiple sclerosis

“…Conceptually, these findings showed that MMPs can not only act as effectors but also regulators of inflammatory responses [47]. Subsequently, CCL7 has been also shown to be a specific substrate of MMP-1, -3, -13 and -14 but not MMP-8 and -9 [48]. The closely related chemokines CCL2, CCL8 and CCL13 (MCP-1, -2 and -4) are proteolytically cleaved by MMP-1 and -3 (but not MMP-2 and -14) with the truncated products of CCL8 and CCL13 being potent antagonists of the their respective chemokine receptors [48].…”

“…Subsequently, CCL7 has been also shown to be a specific substrate of MMP-1, -3, -13 and -14 but not MMP-8 and -9 [48]. The closely related chemokines CCL2, CCL8 and CCL13 (MCP-1, -2 and -4) are proteolytically cleaved by MMP-1 and -3 (but not MMP-2 and -14) with the truncated products of CCL8 and CCL13 being potent antagonists of the their respective chemokine receptors [48]. Apart from influencing the activity of C-C motif chemokines, MMPs also contribute to CXC-chemokine function.…”

Monocyte chemoattractant protein-1 and the blood–brain barrier

“…Which enzyme(s) is responsible for this cleavage, however, is not clear. In the N-terminus, human MCP1 has been reported to be cleaved by matrix metalloproteinase-1, −3, −8, and −12 between aminoacid 4 and 5 (Dean et al, 2008;McQuibban et al, 2002). This cleavage generates a fragment (5-76) that functions as an antagonist for CCR2 (Dean et al, 2008;Gong and Clark-Lewis, 1995;McQuibban et al, 2002;Proost et al, 1998).…”

“…In the N-terminus, human MCP1 has been reported to be cleaved by matrix metalloproteinase-1, −3, −8, and −12 between aminoacid 4 and 5 (Dean et al, 2008;McQuibban et al, 2002). This cleavage generates a fragment (5-76) that functions as an antagonist for CCR2 (Dean et al, 2008;Gong and Clark-Lewis, 1995;McQuibban et al, 2002;Proost et al, 1998). Consistently, the MCP1 mutant lacking amino acids 2-8 (7ND) has been shown to inhibit MCP1-CCR2 signaling both in vitro and in vivo (Kitamoto and Egashira, 2003;Ni et al, 2001;Zhang and Rollins, 1995).…”

The paradox of matrix metalloproteinases in infectious disease

“…The full number of interactions described between MMPs and chemokines is beyond the scope of this review. This mechanism of antagonist-producing chemokine degradation may provide an elegant negative feedback loop that dampens inflammatory cell influx and permits the timely resolution of inflammation [30]. MMP-7 knock-out mice demonstrate a deficit of neutrophil migration that led to the identification of a third mechanism whereby MMPs regulate leucocyte recruitment.…”