CCL5-enhanced human immature dendritic cell migration through the basement membrane in vitro depends on matrix metalloproteinase-9.
The proinflammatory chemokine CC chemokine ligand 5 (CCL5) is a potent chemoattractant of immature dendritic cells (iDCs). It remains to be elucidated whether CCL5 may also enhance iDC migration through the basement membrane by affecting matrix metalloproteinase (MMP)-9 secretion. In this study, iDCs were differentiated in vitro from human monocytes of healthy donors. Zymographic analysis of cellular membranes of nontreated iDCs revealed a basal secretion of the pro- and active MMP-9, whereas only pro-MMP-9 was detected in conditioned media. Increasing concentrations of CCL5 significantly enhanced MMP-9 secretion by iDCs, peaking at 100 ng/ml, which optimally increased iDC migration through a reconstituted basement membrane (Matrigel) in vitro. The CCL5-enhanced secretion of MMP-9 occurred early (2 h) and was maintained at least for 10 h. A significant increase in MMP-9 mRNA synthesis was detected by reverse transcriptase-polymerase chain reaction, only at 6 h of CCL5 treatment, which suggests that the early effect of CCL5 (0-4 h) on MMP-9 secretion was independent of mRNA synthesis, whereas the more delayed effect (6-10 h) could be mediated through an increase in MMP-9 gene expression. In a Matrigel migration assay, the CCL5-enhanced iDC migration was reduced significantly by specific inhibitors of MMP-9, such as tissue inhibitor of metalloproteinase-1 or an anti-MMP-9 antibody, which indicates that iDC migration through the basement membrane depends on MMP-9. These results suggest that under inflammatory conditions, the chemokine CCL5 may enhance iDC migration through the basement membrane by rapidly increasing their MMP-9 secretion.
Chabot
Valérie
V
Etablissement Français du Sang Centre Atlantique, Service Recherche, 2 boulevard Tonnellé, 37020 Tours Cedex, France. valerie.chabot@efs.sante.fr
Reverdiau
Pascale
P
Iochmann
Sophie
S
Rico
Angélique
A
Sénécal
Delphine
D
Goupille
Caroline
C
Sizaret
Pierre-Yves
PY
Sensebé
Luc
L
D000906:Antibodies / Q000494:pharmacology; D001485:Basement Membrane / Q000276:immunology / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000276:immunology; D002465:Cell Movement / Q000187:drug effects / Q000276:immunology*; D018946:Chemokine CCL5; D019742:Chemokines, CC / Q000037:antagonists & inhibitors / Q000276:immunology / Q000494:pharmacology*; D003094:Collagen / Q000276:immunology; D003713:Dendritic Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D006801:Humans; D066298:In Vitro Techniques; D007797:Laminin / Q000276:immunology; D020780:Matrix Metalloproteinase 9 / Q000187:drug effects / Q000378:metabolism*; D009000:Monocytes / Q000187:drug effects / Q000276:immunology; D011509:Proteoglycans / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000187:drug effects; D012016:Reference Values; D013997:Time Factors; D019714:Tissue Inhibitor of Metalloproteinases / Q000494:pharmacology
D000906:Antibodies; C483024:CCL5 protein, human; D018946:Chemokine CCL5; D019742:Chemokines, CC; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; C059485:matrigel; D003094:Collagen; D020780:Matrix Metalloproteinase 9