1. Flow cytometry and gene expression profiling of immune cells of the carotid plaque and peripheral blood.
- Author
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Sternberg Z, Ghanim H, Gillotti KM, Tario JD Jr, Munschauer F, Curl R, Noor S, Yu J, Ambrus JL Sr, Wallace P, and Dandona P
- Subjects
- Aged, Aged, 80 and over, B-Lymphocytes cytology, B-Lymphocytes immunology, Cell Adhesion immunology, Female, Flow Cytometry, Humans, Immunomagnetic Separation, Immunophenotyping, Male, Middle Aged, Monocytes cytology, Monocytes immunology, T-Lymphocytes cytology, T-Lymphocytes immunology, Thrombosis genetics, Thrombosis immunology, Thrombosis pathology, Carotid Stenosis genetics, Carotid Stenosis immunology, Carotid Stenosis pathology, Leukocytes, Mononuclear cytology, Leukocytes, Mononuclear immunology, Transcriptome immunology, Vasculitis genetics, Vasculitis immunology, Vasculitis pathology
- Abstract
Objectives: 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., Conclusion: 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., (Published by Elsevier Ireland Ltd.)
- Published
- 2013
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