title: Gene profiling in atherosclerosis reveals a key role for small inducible cytokines: validation using a novel monocyte chemoattractant protein monoclonal antibody.
abstract: BACKGROUND
  Pathological aspects of atherosclerosis are well described, but gene profiles during atherosclerotic plaque progression are largely unidentified.
  
  
  RESULTS
  Microarray analysis was performed on mRNA of aortic arches of ApoE-/- mice fed normal chow (NC group) or Western-type diet (WD group) for 3, 4.5, and 6 months. Of 10 176 reporters, 387 were differentially (>2x) expressed in at least 1 group compared with a common reference (ApoE-/-, 3- month NC group). The number of differentially expressed genes increased during plaque progression. Time-related expression clustering and functional grouping of differentially expressed genes suggested important functions for genes involved in inflammation (especially the small inducible cytokines monocyte chemoattractant protein [MCP]-1, MCP-5, macrophage inflammatory protein [MIP]-1alpha, MIP-1beta, MIP-2, and fractalkine) and matrix degradation (cathepsin-S, matrix metalloproteinase-2/12). Validation experiments focused on the gene cluster of small inducible cytokines. Real-time polymerase chain reaction revealed a plaque progression-dependent increase in mRNA levels of MCP-1, MCP-5, MIP-1alpha, and MIP-1beta. ELISA for MCP-1 and MCP-5 showed similar results. Immunohistochemistry for MCP-1, MCP-5, and MIP-1alpha located their expression to plaque macrophages. An inhibiting antibody for MCP-1 and MCP-5 (11K2) was designed and administered to ApoE-/- mice for 12 weeks starting at the age of 5 or 17 weeks. 11K2 treatment reduced plaque area and macrophage and CD45+ cell content and increased collagen content, thereby inducing a stable plaque phenotype.
  
  
  CONCLUSIONS
  Gene profiling of atherosclerotic plaque progression in ApoE-/- mice revealed upregulation of the gene cluster of small inducible cytokines. Further expression and in vivo validation studies showed that this gene cluster mediates plaque progression and stability.
authors.affiliation: Department of Pathology, Cardiovascular Research Institute Maastricht, University of Maastricht, Maastricht, The Netherlands. e.lutgens@path.unimaas.nl
authors.forename: Esther
authors.initials: E
authors.lastname: Lutgens
authors.forename: Birgit
authors.initials: B
authors.lastname: Faber
authors.forename: Kitty
authors.initials: K
authors.lastname: Schapira
authors.forename: Chris T A
authors.initials: CT
authors.lastname: Evelo
authors.forename: Rachel
authors.initials: R
authors.lastname: van Haaften
authors.forename: Sylvia
authors.initials: S
authors.lastname: Heeneman
authors.forename: Kitty B J M
authors.initials: KB
authors.lastname: Cleutjens
authors.forename: Ann Pascale
authors.initials: AP
authors.lastname: Bijnens
authors.forename: Linda
authors.initials: L
authors.lastname: Beckers
authors.forename: J Gordon
authors.initials: JG
authors.lastname: Porter
authors.forename: Charles R
authors.initials: CR
authors.lastname: Mackay
authors.forename: Paul
authors.initials: P
authors.lastname: Rennert
authors.forename: Veronique
authors.initials: V
authors.lastname: Bailly
authors.forename: Matthew
authors.initials: M
authors.lastname: Jarpe
authors.forename: Brian
authors.initials: B
authors.lastname: Dolinski
authors.forename: Victor
authors.initials: V
authors.lastname: Koteliansky
authors.forename: Tony
authors.initials: T
authors.lastname: de Fougerolles
authors.forename: Mat J A P
authors.initials: MJ
authors.lastname: Daemen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000494:pharmacology; D001013:Aorta, Thoracic; D001057:Apolipoproteins E / Q000172:deficiency; D050197:Atherosclerosis / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D018932:Chemokine CCL2 / Q000276:immunology; D054412:Chemokine CCL8; D018925:Chemokines / Q000235:genetics / Q000502:physiology*; D016000:Cluster Analysis; D018450:Disease Progression; D005109:Extracellular Matrix / Q000378:metabolism; D020869:Gene Expression Profiling; D007249:Inflammation / Q000235:genetics; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D018945:Monocyte Chemoattractant Proteins / Q000276:immunology / Q000502:physiology; D010447:Peptide Hydrolases / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D001057:Apolipoproteins E; C491768:Ccl2 protein, mouse; C515454:Ccl8 protein, mouse; D018932:Chemokine CCL2; D054412:Chemokine CCL8; D018925:Chemokines; D018945:Monocyte Chemoattractant Proteins; D012333:RNA, Messenger; D010447:Peptide Hydrolases