Fluorescent activated cell sorting: An effective approach to study dendritic cell subsets in human atherosclerotic plaques

• Published in: Journal of immunological methods 2015-02, Vol.417, p.76-85
• Main Authors: Van Brussel, Ilse, Ammi, Rachid, Rombouts, Miche, Cools, Nathalie, Vercauteren, Sven R., De Roover, Dominique, Hendriks, Jeroen M.H., Lauwers, Patrick, Van Schil, Paul E., Schrijvers, Dorien M.
• Format: Article
• Language: English
• Subjects: Aged; Aged, 80 and over; Atherosclerosis; Atherosclerosis - immunology; B-Lymphocytes - cytology; CD11b Antigen - metabolism; Cell Separation - methods; Dendritic cells; Dendritic Cells - immunology; Female; Flow Cytometry - methods; Fluorescence activated cell sorting; GPI-Linked Proteins - metabolism; Humans; Inflammation - immunology; Interleukin-3 Receptor alpha Subunit - metabolism; Killer Cells, Natural - cytology; Lectins, C-Type - metabolism; Macrophages - cytology; Male; Middle Aged; Monocytes - cytology; Plaque, Atherosclerotic - immunology; Plaques; Receptors, IgG - metabolism; Receptors, Mitogen - metabolism; T-Lymphocytes - cytology
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/j.jim.2014.12.010

Different immune cell types are present within atherosclerotic plaques. Dendritic cells (DC) are of special interest, since they are considered as the ‘center of the immuniverse’. Identifying inflammatory DC subtypes within plaques is important for a better understanding of the lesion pathogenesis and pinpoints their contribution to the atherosclerotic process. We have developed a flow cytometry-based method to characterize and isolate different DC subsets (i.e. CD11b+, Clec9A+ and CD16+ conventional (c)DC and CD123+ plasmacytoid (p)DC) in human atherosclerotic plaques. We revealed a predominance of pro-inflammatory CD11b+ DC in advanced human lesions, whereas atheroprotective Clec9A+ DC were almost absent. CD123+ pDC and CD16+ DC were also detectable in plaques. Remarkably, plaques from distinct anatomical locations exhibited different cellular compositions: femoral plaques contained less CD11b+ and Clec9A+ DC than carotid plaques. Twice as many monocytes/macrophages were observed compared to DC. Moreover, relative amounts of T cells/B cells/NK cells were 6 times as high as DC numbers. For the first time, fluorescent activated cell sorting analysis of DC subsets in human plaques indicated a predominance of CD11b+ cDC, in comparison with other DC subsets. Isolation of the different subsets will facilitate detailed functional analysis and may have significant implications for tailoring appropriate therapy. •Understanding the origin/function of DC subsets that control atherogenesis is needed.•It can open up strategies to interfere with DC subset differentiation and functions for developing therapeutic targets.•FACS allowed us to characterize the presence of specialized DC subsets in human atherosclerotic lesions.•FACS analysis of carotid and femoral arteries revealed a predominance of the CD11bpos subset.•Also, different locations in the vascular tree show differences in the accumulation of specialized DC subsets.