title: Fluorescent activated cell sorting: an effective approach to study dendritic cell subsets in human atherosclerotic plaques.
abstract: 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.
authors.affiliation: Lab of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium. Electronic address: ilse.vanbrussel@uantwerpen.be.
authors.forename: Ilse
authors.initials: I
authors.lastname: Van Brussel
authors.affiliation: Lab of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium.
authors.forename: Rachid
authors.initials: R
authors.lastname: Ammi
authors.affiliation: Lab of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium.
authors.forename: Miche
authors.initials: M
authors.lastname: Rombouts
authors.affiliation: Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (VAXINFECTIO), University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Cools
authors.affiliation: Department of Thoracic and Vascular Surgery, ZNA Middelheim (Ziekenhuis Netwerk Antwerpen), Lindendreef 1, B-2020 Antwerp, Belgium.
authors.forename: Sven R
authors.initials: SR
authors.lastname: Vercauteren
authors.affiliation: Department of Thoracic and Vascular Surgery, ZNA Middelheim (Ziekenhuis Netwerk Antwerpen), Lindendreef 1, B-2020 Antwerp, Belgium.
authors.forename: Dominique
authors.initials: D
authors.lastname: De Roover
authors.affiliation: Department of Thoracic and Vascular Surgery, Antwerp University Hospital (UZA), Wilrijkstraat 10, B-2650 Edegem, Antwerp, Belgium.
authors.forename: Jeroen M H
authors.initials: JMH
authors.lastname: Hendriks
authors.affiliation: Department of Thoracic and Vascular Surgery, Antwerp University Hospital (UZA), Wilrijkstraat 10, B-2650 Edegem, Antwerp, Belgium.
authors.forename: Patrick
authors.initials: P
authors.lastname: Lauwers
authors.affiliation: Department of Thoracic and Vascular Surgery, Antwerp University Hospital (UZA), Wilrijkstraat 10, B-2650 Edegem, Antwerp, Belgium.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Van Schil
authors.affiliation: Lab of Physiopharmacology, University of Antwerp, Universiteitsplein 1, B-2610 Antwerp, Belgium.
authors.forename: Dorien M
authors.initials: DM
authors.lastname: Schrijvers
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D050197:Atherosclerosis / Q000276:immunology*; D001402:B-Lymphocytes / Q000166:cytology; D039481:CD11b Antigen / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D003713:Dendritic Cells / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D058851:GPI-Linked Proteins / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000276:immunology; D053650:Interleukin-3 Receptor alpha Subunit / Q000378:metabolism; D007694:Killer Cells, Natural / Q000166:cytology; D037181:Lectins, C-Type / Q000378:metabolism; D008264:Macrophages / Q000166:cytology; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000166:cytology; D058226:Plaque, Atherosclerotic / Q000276:immunology*; D017452:Receptors, IgG / Q000378:metabolism; D011975:Receptors, Mitogen / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology
chemical_list: D039481:CD11b Antigen; C530206:CLEC9a protein, human; C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; C115022:IL3RA protein, human; C509295:ITGAM protein, human; D053650:Interleukin-3 Receptor alpha Subunit; D037181:Lectins, C-Type; D017452:Receptors, IgG; D011975:Receptors, Mitogen
keywords: Atherosclerosis; Dendritic cells; Fluorescence activated cell sorting; Plaques