title: Novel proteolytic microvesicles released from human macrophages after exposure to tobacco smoke.
abstract: Cigarette smoking damages the extracellular matrix in a variety of locations, leading to atherosclerotic plaque instability and emphysematous lung destruction, but the underlying mechanisms remain poorly understood. Here, we sought to determine whether exposure of human macrophages, a key participant in extracellular matrix damage, to tobacco smoke extract (TSE) induces the release of microvesicles (MVs; or microparticles) with proteolytic activity; the major proteases involved; and the cellular mechanisms that might mediate their generation. We found that MVs released from TSE-exposed macrophages carry substantial gelatinolytic and collagenolytic activities that surprisingly can be predominantly attributed to a single transmembrane protease of the matrix metalloproteinase (MMP) superfamily (namely, MMP14). Flow cytometric counts revealed that exposure of human macrophages to TSE for 20 hours more than quadrupled their production of MMP14-positive MVs (control, 1112 ± 231; TSE-induced, 5823 ± 2192 MMP14-positive MVs/μL of conditioned medium; means ± SEM; n = 6; P < 0.01). Our results indicate that the production of these MVs by human macrophages relies on a series of regulated steps that include activation of two mitogen-activated protein kinases (MAPKs, i.e., the Jun N-terminal kinase and p38 MAPK), and then MAPK-dependent induction and maturation of cellular MMP14, a remarkable accumulation of MMP14 into nascent plasma membrane blebs, and finally caspase- and MAPK-dependent apoptosis and apoptotic microvesicle generation. Proteolytically active MVs induced by tobacco smoke may be novel mediators of clinical important matrix destruction in smokers.
authors.affiliation: Section of Endocrinology, Diabetes, and Metabolism, Department of Medicine, Temple University School of Medicine, Philadelphia, PA 19140, USA.
authors.forename: Chun-Jun
authors.initials: CJ
authors.lastname: Li
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.forename: Demin
authors.initials: D
authors.lastname: Yu
authors.forename: Kevin Jon
authors.initials: KJ
authors.lastname: Williams
authors.forename: Ming-Lin
authors.initials: ML
authors.lastname: Liu
mesh_terms: D017209:Apoptosis; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008264:Macrophages / Q000201:enzymology / Q000378:metabolism*; D053511:Matrix Metalloproteinase 14 / Q000378:metabolism; D059748:Proteolysis; D012907:Smoking; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C507086:MMP14 protein, human; D053511:Matrix Metalloproteinase 14