title: Macrophage activation by heparanase is mediated by TLR-2 and TLR-4 and associates with plaque progression.
abstract: OBJECTIVE
  Factors and mechanisms that activate macrophages in atherosclerotic plaques are incompletely understood. We examined the capacity of heparanase to activate macrophages.
  
  
  RESULTS
  Highly purified heparanase was added to mouse peritoneal macrophages and macrophage-like J774 cells, and the levels of tumor necrosis factor-α, matrix metalloproteinase-9, interlukin-1, and monocyte chemotactic protein-1 were evaluated by ELISA. Gene expression was determined by RT-PCR. Cells collected from Toll-like receptor-2 and Toll-like receptor-4 knockout mice were evaluated similarly. Heparanase levels in the plasma of patients with acute myocardial infarction, stable angina, and healthy subjects were determined by ELISA. Immunohistochemistry was applied to detect the expression of heparanase in control specimens and specimens of patients with stable angina or acute myocardial infarction. Addition or overexpression of heparanase variants resulted in marked increase in tumor necrosis factor-α, matrix metalloproteinase-9, interlukin-1, and monocyte chemotactic protein-1 levels. Mouse peritoneal macrophages harvested from Toll-like receptor-2 or Toll-like receptor-4 knockout mice were not activated by heparanase. Plasma heparanase level was higher in patients with acute myocardial infarction, compared with patients with stable angina and healthy subjects. Pathologic coronary specimens obtained from vulnerable plaques showed increased heparanase staining compared with specimens of stable plaque and controls.
  
  
  CONCLUSIONS
  Heparanase activates macrophages, resulting in marked induction of cytokine expression associated with plaque progression toward vulnerability.
authors.affiliation: Department of Cardiology and Pathology, Rambam Health Care Campus, Haifa, Israel.
authors.forename: Miry
authors.initials: M
authors.lastname: Blich
authors.forename: Amnon
authors.initials: A
authors.lastname: Golan
authors.forename: Gil
authors.initials: G
authors.lastname: Arvatz
authors.forename: Anat
authors.initials: A
authors.lastname: Sebbag
authors.forename: Itay
authors.initials: I
authors.lastname: Shafat
authors.forename: Edmond
authors.initials: E
authors.lastname: Sabo
authors.forename: Victoria
authors.initials: V
authors.lastname: Cohen-Kaplan
authors.forename: Sirouch
authors.initials: S
authors.lastname: Petcherski
authors.forename: Shani
authors.initials: S
authors.lastname: Avniel-Polak
authors.forename: Amnon
authors.initials: A
authors.lastname: Eitan
authors.forename: Haim
authors.initials: H
authors.lastname: Hammerman
authors.forename: Doron
authors.initials: D
authors.lastname: Aronson
authors.forename: Elena
authors.initials: E
authors.lastname: Axelman
authors.forename: Neta
authors.initials: N
authors.lastname: Ilan
authors.forename: Gabriel
authors.initials: G
authors.lastname: Nussbaum
authors.forename: Israel
authors.initials: I
authors.lastname: Vlodavsky
mesh_terms: D060050:Angina, Stable / Q000097:blood / Q000201:enzymology; D000818:Animals; D050197:Atherosclerosis / Q000201:enzymology* / Q000235:genetics / Q000276:immunology / Q000473:pathology; D002460:Cell Line; D018932:Chemokine CCL2 / Q000378:metabolism; D003324:Coronary Artery Disease / Q000097:blood / Q000201:enzymology; D018450:Disease Progression; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D005966:Glucuronidase / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000378:metabolism; D008262:Macrophage Activation; D017737:Macrophages, Peritoneal / Q000201:enzymology* / Q000276:immunology / Q000473:pathology; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009203:Myocardial Infarction / Q000097:blood / Q000201:enzymology; D058226:Plaque, Atherosclerotic; D016133:Polymerase Chain Reaction; D012422:Rupture, Spontaneous; D015398:Signal Transduction; D013997:Time Factors; D051195:Toll-Like Receptor 2 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000099175:Heparanase
chemical_list: D018932:Chemokine CCL2; D005966:Glucuronidase; D007375:Interleukin-1; D020780:Matrix Metalloproteinase 9; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D000099175:Heparanase; C491768:Ccl2 protein, mouse; C493486:Tlr2 protein, mouse; C493487:Tlr4 protein, mouse; C501830:Mmp9 protein, mouse