title: Dynamic immune cell accumulation during flow-induced atherogenesis in mouse carotid artery: an expanded flow cytometry method.
abstract: OBJECTIVE
  Inflammation plays a central role in atherosclerosis. However, the detailed changes in the composition and quantity of leukocytes in the arterial wall during atherogenesis are not fully understood in part because of the lack of suitable methods and animal models.
  
  
  RESULTS
  We developed a 10-fluorochrome, 13-parameter flow cytometry method to quantitate 7 major leukocyte subsets in a single digested arterial wall sample. Apolipoprotein E-deficient mice underwent left carotid artery (LCA) partial ligation and were fed a high-fat diet for 4 to 28 days. Monocyte/macrophages, dendritic cells, granulocytes, natural killer cells, and CD4 T cells significantly infiltrated the LCA as early as 4 days. Monocyte/macrophages and dendritic cells decreased between 7 and 14 days, whereas T-cell numbers remained steady. Leukocyte numbers peaked at 7 days, preceding atheroma formation at 14 days. B cells entered LCA by 14 days. Control right carotid and sham-ligated LCAs showed no significant infiltrates. Polymerase chain reaction and ELISA arrays showed that expression of proinflammatory cytokines and chemokines peaked at 7 and 14 days postligation, respectively.
  
  
  CONCLUSIONS
  This is the first quantitative description of leukocyte number and composition over the life span of murine atherosclerosis. These results show that disturbed flow induces rapid and dynamic leukocyte accumulation in the arterial wall during the initiation and progression of atherosclerosis.
authors.affiliation: School of Medicine, Emory University, Woodruff Memorial Bldg, Rm 2005, Atlanta, GA 30322, USA.
authors.forename: Noah
authors.initials: N
authors.lastname: Alberts-Grill
authors.forename: Amir
authors.initials: A
authors.lastname: Rezvan
authors.forename: Dong Ju
authors.initials: DJ
authors.lastname: Son
authors.forename: Haiwei
authors.initials: H
authors.lastname: Qiu
authors.forename: Chan Woo
authors.initials: CW
authors.lastname: Kim
authors.forename: Melissa L
authors.initials: ML
authors.lastname: Kemp
authors.forename: Cornelia M
authors.initials: CM
authors.lastname: Weyand
authors.forename: Hanjoong
authors.initials: H
authors.lastname: Jo
mesh_terms: D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency / Q000235:genetics; D002339:Carotid Arteries / Q000276:immunology* / Q000503:physiopathology / Q000601:surgery; D002340:Carotid Artery Diseases / Q000209:etiology / Q000235:genetics / Q000276:immunology* / Q000503:physiopathology; D018925:Chemokines / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D059305:Diet, High-Fat; D004195:Disease Models, Animal; D018450:Disease Progression; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D007113:Immunity, Innate; D016130:Immunophenotyping / Q000379:methods*; D018836:Inflammation Mediators / Q000378:metabolism; D007958:Leukocyte Count; D007962:Leukocytes / Q000145:classification / Q000276:immunology*; D008026:Ligation; D051379:Mice; D018345:Mice, Knockout; D016133:Polymerase Chain Reaction; D012039:Regional Blood Flow; D013997:Time Factors
chemical_list: D001057:Apolipoproteins E; D018925:Chemokines; D016207:Cytokines; D018836:Inflammation Mediators