title: Flow cytometry and gene expression profiling of immune cells of the carotid plaque and peripheral blood.
abstract: OBJECTIVE
  The relative contribution of the local vs. peripheral inflammation to the atherothrombotic processes is unknown. We compared the inflammatory status of the immune cells of the carotid plaque with similar cells in peripheral circulation of patients with advanced carotid disease (PCDs).
  
  
  METHODS
  Mononuclear cells (MNCs) were extracted from carotid endarterectomy (CEA) samples by enzymatic digestion and subsequent magnetic cell sorting. The cell surface antigenic expressions, and mRNA expression levels were compared between CEA MNCs and peripheral MNCs, using flow cytometry and RT-PCR techniques.
  
  
  RESULTS
  The percentages of resting MNCs were lower, and activated MNCs, particularly monocytes, were higher in the CEAMNCs, as compared to the peripheral MNCs. The percentages of activated T cells and B cells were higher in the peripheral MNCs of PCDs, than in healthy controls (HCs), but the percentages of activated monocytes did not differ between the two groups. The expression levels of both pro-inflammatory/pro-thrombotic (P(38), JNKB-1, Egr-1 PAI-1, MCP-1, TF, MMP-9, HMGB-1, TNF-α, mTOR) and anti-inflammatory (PPAR-γ, TGF-β) mediators were significantly higher in the CEA MNCs as compared to the peripheral MNCs. Furthermore, MMP-9 and PPAR-γ expression levels were higher in the peripheral MNCs of PCDs than HCs.
  
  
  CONCLUSIONS
  The inflammatory status is higher in the immune cells of the carotid plaque, as compared to those cells in the peripheral blood. The altered expression levels of both pro-inflammatory/pro-thrombotic and anti-inflammatory mediators in the milieu of the plaque suggest that the balance between these various mediators may play a key role in carotid disease progression.
authors.affiliation: Stroke Center, Kaleida Medical Center, Buffalo, NY, USA. zs2@buffalo.edu
authors.forename: Zohara
authors.initials: Z
authors.lastname: Sternberg
authors.forename: Husam
authors.initials: H
authors.lastname: Ghanim
authors.forename: Kristen M
authors.initials: KM
authors.lastname: Gillotti
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Tario
authors.forename: Frederick
authors.initials: F
authors.lastname: Munschauer
authors.forename: Richard
authors.initials: R
authors.lastname: Curl
authors.forename: Sonya
authors.initials: S
authors.lastname: Noor
authors.forename: Jihnhee
authors.initials: J
authors.lastname: Yu
authors.forename: Julian L
authors.initials: JL
authors.lastname: Ambrus
authors.forename: Paul
authors.initials: P
authors.lastname: Wallace
authors.forename: Paresh
authors.initials: P
authors.lastname: Dandona
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D016893:Carotid Stenosis / Q000235:genetics / Q000276:immunology / Q000473:pathology; D002448:Cell Adhesion / Q000276:immunology; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D018189:Immunomagnetic Separation; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D013927:Thrombosis / Q000235:genetics / Q000276:immunology / Q000473:pathology; D059467:Transcriptome / Q000276:immunology*; D014657:Vasculitis / Q000235:genetics / Q000276:immunology / Q000473:pathology
keywords: ATF; BMI; BP; CAD; CDP; CEA; CV; CVD; Carotid stenosis; Cell adhesion; Cell surface markers; DM; ERK; Egr; Gene expression; HC; HF; HMGB; Immunophenotyping; JNK; Jun N-terminal kinase; MAP; MCP; MMP; MNC; MRA; Mononuclear cells; NF-KB; P; PAD; PAI; PMT; PPAR; TF; TGF; TLR; TNF; Thrombosis; VCAM; atrial fibrillation; blood pressure; body mass index; cardiovascular; carotid disease patients; carotid endarterectomy; cerebrovascular disease; coronary artery disease; diabetes mellitus; early growth response; extracellular regulated kinase; healthy control; heart failure; high-mobility group protein B; mTOR; magnetic resonance angiography; mammalian target of rapamycin; matrix metalloproteinase; mitogen activated protein; monocyte chemotactic protein; mononuclear cell; nuclear factor kappa B; peripheral; peripheral arterial disease; peroxisome proliferator-activated receptor; photomultipler tube; plasminogen activator inhibitor; tissue factor; toll-like receptor; transforming growth factor; tumor necrosis factor; vascular cell adhesion molecule