title: Monocyte ADAM17 promotes diapedesis during transendothelial migration: identification of steps and substrates targeted by metalloproteinases.
abstract: Despite expanded definition of the leukocyte adhesion cascade and mechanisms underlying individual steps, very little is known about regulatory mechanisms controlling sequential shifts between steps. We tested the hypothesis that metalloproteinases provide a mechanism to rapidly transition monocytes between different steps. Our study identifies diapedesis as a step targeted by metalloproteinase activity. Time-lapse video microscopy shows that the presence of a metalloproteinase inhibitor results in a doubling of the time required for human monocytes to complete diapedesis on unactivated or inflamed human endothelium, under both static and physiological-flow conditions. Thus, diapedesis is promoted by metalloproteinase activity. In contrast, neither adhesion of monocytes nor their locomotion over the endothelium is altered by metalloproteinase inhibition. We further demonstrate that metalloproteinase inhibition significantly elevates monocyte cell surface levels of integrins CD11b/CD18 (Mac-1), specifically during transendothelial migration. Interestingly, such alterations are not detected for other endothelial- and monocyte-adhesion molecules that are presumed metalloproteinase substrates. Two major transmembrane metalloproteinases, a disintegrin and metalloproteinase (ADAM)17 and ADAM10, are identified as enzymes that control constitutive cleavage of Mac-1. We further establish that knockdown of monocyte ADAM17, but not endothelial ADAM10 or ADAM17 or monocyte ADAM10, reproduces the diapedesis delay observed with metalloproteinase inhibition. Therefore, we conclude that monocyte ADAM17 facilitates the completion of transendothelial migration by accelerating the rate of diapedesis. We propose that the progression of diapedesis may be regulated by spatial and temporal cleavage of Mac-1, which is triggered upon interaction with endothelium.
authors.affiliation: Department of Pathology, University of Washington School of Medicine, Seattle, WA 98104, USA.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Tsubota
authors.forename: Jeremy M
authors.initials: JM
authors.lastname: Frey
authors.forename: Phillip W L
authors.initials: PW
authors.lastname: Tai
authors.forename: Robert E
authors.initials: RE
authors.lastname: Welikson
authors.forename: Elaine W
authors.initials: EW
authors.lastname: Raines
mesh_terms: D051722:ADAM Proteins / Q000172:deficiency / Q000378:metabolism / Q000502:physiology*; D000072198:ADAM17 Protein; D002478:Cells, Cultured; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D016177:Macrophage-1 Antigen / Q000378:metabolism; D045726:Metalloproteases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009000:Monocytes / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D013379:Substrate Specificity / Q000276:immunology; D059008:Time-Lapse Imaging / Q000379:methods; D057705:Transendothelial and Transepithelial Migration / Q000276:immunology*
chemical_list: D016177:Macrophage-1 Antigen; D045726:Metalloproteases; D051722:ADAM Proteins; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human