title: Translocation of active heparanase to cell surface regulates degradation of extracellular matrix heparan sulfate upon transmigration of mature monocyte-derived dendritic cells.
abstract: After Ag capture and exposure to danger stimuli, maturing dendritic cells (DCs) migrate to regional lymph nodes, where antigenic peptides are presented to T lymphocytes. To migrate from peripheral tissue such as the epidermis to regional lymph nodes, Ag-bearing epidermal Langerhans cells must move through an extracellular matrix (ECM) of various compositions. The nature of their capacity to transmigrate via ECM is not well understood, although MIP-3beta and CCR7 play critical roles. We were interested in verifying whether heparanase, a heparan sulfate-degrading endo-beta-d-glucuronidase that participates in ECM degradation and remodeling, is expressed and functional in monocyte-derived DCs. Using immunohistochemistry, confocal microscopy, RT-PCR, Western blot analysis, assays for heparanase activity, and Matrigel transmigration, we show that heparanase is expressed in both nuclei and cytoplasm of immature DCs, and that gene expression and synthesis take place mainly in monocytes and early immature DCs. We also found that both nuclear and cytoplasm fractions show heparanase activity, and upon LPS-induced maturation, heparanase translocates to the cell surface and degrades ECM heparan sulfate. Matrigel transmigration assays showed a MIP-3beta-comparable role for heparanase. Because heparan sulfate glycosaminoglycans play a key role in the self-assembly, insolubility, and barrier properties of the ECM, the results of this study suggest that heparanase is a key enzyme in DC transmigration through the ECM.
authors.affiliation: Laboratory for Cellular and Molecular Immunology, Bruce Rappaport Faculty of Medicine, Technion, Israel.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Benhamron
authors.forename: Hovav
authors.initials: H
authors.lastname: Nechushtan
authors.forename: Inna
authors.initials: I
authors.lastname: Verbovetski
authors.forename: Alon
authors.initials: A
authors.lastname: Krispin
authors.forename: Ghada
authors.initials: G
authors.lastname: Abboud-Jarrous
authors.forename: Eyal
authors.initials: E
authors.lastname: Zcharia
authors.forename: Evgeny
authors.initials: E
authors.lastname: Edovitsky
authors.forename: Efrat
authors.initials: E
authors.lastname: Nahari
authors.forename: Tamar
authors.initials: T
authors.lastname: Peretz
authors.forename: Israel
authors.initials: I
authors.lastname: Vlodavsky
authors.forename: Dror
authors.initials: D
authors.lastname: Mevorach
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002454:Cell Differentiation / Q000276:immunology; D002465:Cell Movement / Q000276:immunology*; D002467:Cell Nucleus / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D003713:Dendritic Cells / Q000166:cytology / Q000201:enzymology*; D004728:Endothelium, Corneal / Q000166:cytology / Q000201:enzymology* / Q000276:immunology; D004789:Enzyme Activation; D005109:Extracellular Matrix / Q000201:enzymology / Q000378:metabolism*; D005966:Glucuronidase / Q000096:biosynthesis / Q000378:metabolism* / Q000502:physiology; D006497:Heparan Sulfate / Q000378:metabolism*; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology; D008297:Male; D008565:Membrane Proteins / Q000502:physiology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000201:enzymology*; D021381:Protein Transport / Q000276:immunology; D000099175:Heparanase
chemical_list: D005966:Glucuronidase; D006497:Heparan Sulfate; D008565:Membrane Proteins; D000099175:Heparanase