title: Delta-like 4 induces notch signaling in macrophages: implications for inflammation.
abstract: BACKGROUND
  Activated macrophages contribute to the pathogenesis of inflammatory diseases such as atherosclerosis. Although Notch signaling participates in various aspects of immunity, its role in macrophage activation remains undetermined.
  
  
  RESULTS
  To explore the role of Notch signaling in inflammation, we examined the expression and activity of Notch pathway components in human primary macrophages in vitro and in atherosclerotic plaques. Macrophages in culture express various Notch pathway components including all 4 receptors (Notch1 to Notch4). Notch3 selectively increased during macrophage differentiation; however, silencing by RNA interference demonstrated that all receptors are functional. The ligand Delta-like 4 (Dll4) increased in macrophages exposed to proinflammatory stimuli such as lipopolysaccharide, interleukin-1beta, or minimally-modified low-density lipoprotein in a Toll-like receptor 4- and nuclear factor-kappaB-dependent fashion. Soluble Dll4 bound to human macrophages. Coincubation of macrophages with cells that expressed Dll4 triggered Notch proteolysis and activation; increased the transcription of proinflammatory genes such as inducible nitric oxide synthase, pentraxin 3 and Id1; resulted in activation of mitogen-activated protein kinase, Akt, and nuclear factor-kappaB pathways; and increased the expression of Dll4 in macrophages. Notch3 knockdown during macrophage differentiation decreased the transcription of genes that promote inflammation, such as inducible nitric oxide synthase, pentraxin 3, Id1, and scavenger receptor-A. These in vitro findings correlate with results of quantitative immunohistochemistry, which demonstrated the presence of Dll4 and other Notch components within macrophages in atherosclerotic plaques.
  
  
  CONCLUSIONS
  Dll4-triggered Notch signaling may mediate inflammatory responses in macrophages and promote inflammation.
authors.affiliation: Center for Excellence in Vascular Biology, Brigham and Women's Hospital, Harvard Medical School, 77 Ave Louis Pasteur, Boston, MA 02115, USA.
authors.forename: Erik
authors.initials: E
authors.lastname: Fung
authors.forename: Sai-Man Timothy
authors.initials: SM
authors.lastname: Tang
authors.forename: James P
authors.initials: JP
authors.lastname: Canner
authors.forename: Kunio
authors.initials: K
authors.lastname: Morishige
authors.forename: Joseph F
authors.initials: JF
authors.lastname: Arboleda-Velasquez
authors.forename: Angelo A
authors.initials: AA
authors.lastname: Cardoso
authors.forename: Nadia
authors.initials: N
authors.lastname: Carlesso
authors.forename: Jon C
authors.initials: JC
authors.lastname: Aster
authors.forename: Masanori
authors.initials: M
authors.lastname: Aikawa
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D050197:Atherosclerosis / Q000473:pathology; D002135:Calcium-Binding Proteins; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation / Q000235:genetics*; D036341:Intercellular Signaling Peptides and Proteins / Q000502:physiology*; D008262:Macrophage Activation; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000378:metabolism* / Q000502:physiology; D015398:Signal Transduction; D014158:Transcription, Genetic
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002135:Calcium-Binding Proteins; C424126:DLL4 protein, human; D036341:Intercellular Signaling Peptides and Proteins; C499374:NOTCH3 protein, human; D000071656:Receptor, Notch3; D051880:Receptors, Notch