title: Multi-analyte profiling in human carotid atherosclerosis uncovers pro-inflammatory macrophage programming in plaques.
abstract: Molecular characterisation of vulnerable atherosclerosis is necessary for targeting functional imaging and plaque-stabilising therapeutics. Inflammation has been linked to atherogenesis and the development of high-risk plaques. We set to quantify cytokine, chemokine and matrix metalloproteinase (MMP) protein production in cells derived from carotid plaques to map the inflammatory milieu responsible for instability. Carotid endarterectomies from carefully characterised symptomatic (n=35) and asymptomatic (n=32) patients were enzymatically dissociated producing mixed cell type atheroma cell suspensions which were cultured for 24 hours. Supernatants were interrogated for 45 analytes using the Luminex 100 platform. Twenty-nine of the 45 analytes were reproducibly detectable in the majority of donors. The in vitro production of a specific network of mediators was found to be significantly higher in symptomatic than asymptomatic plaques, including: tumour necrosis factor α, interleukin (IL) 1β, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), CCL5, CCL20, CXCL9, matrix metalloproteinase (MMP)-3 and MMP-9. Ingenuity pathway analysis of differentially expressed analytes between symptomatic and asymptomatic patients identified a number of key biological pathways (p< 10(-25)). In conclusion, the carotid artery plaque culprit of ischaemic neurological symptoms is characterised by an inflammatory milieu favouring inflammatory cell recruitment and pro-inflammatory macrophage polarisation.
authors.forename: Joseph
authors.initials: J
authors.lastname: Shalhoub
authors.forename: Leena E
authors.initials: LE
authors.lastname: Viiri
authors.forename: Amanda J
authors.initials: AJ
authors.lastname: Cross
authors.forename: Scott M
authors.initials: SM
authors.lastname: Gregan
authors.forename: David M
authors.initials: DM
authors.lastname: Allin
authors.forename: Nagore
authors.initials: N
authors.lastname: Astola
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Franklin
authors.forename: Alun H
authors.initials: AH
authors.lastname: Davies
authors.affiliation: Claudia Monaco, Professor of Cardiovascular Inflammation, Kennedy Institute of Rheumatology, Nuffield Department of Orthopedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Roosevelt Drive, Headington, Oxford OX3 7FY, UK, Tel.: +44 1865 612 636, Fax: +44 1865 612 601, E-mail: claudia.monaco@kennedy.ox.ac.uk.
authors.forename: Claudia
authors.initials: C
authors.lastname: Monaco
mesh_terms: D000368:Aged; D002340:Carotid Artery Diseases / Q000276:immunology / Q000378:metabolism* / Q000601:surgery; D016893:Carotid Stenosis / Q000276:immunology / Q000378:metabolism / Q000601:surgery; D018925:Chemokines / Q000378:metabolism; D003115:Colony-Stimulating Factors / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D016894:Endarterectomy, Carotid; D005260:Female; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism; D008875:Middle Aged; D040081:Protein Array Analysis; D019714:Tissue Inhibitor of Metalloproteinases / Q000378:metabolism
chemical_list: D018925:Chemokines; D003115:Colony-Stimulating Factors; D016207:Cytokines; D018836:Inflammation Mediators; D019714:Tissue Inhibitor of Metalloproteinases; D020782:Matrix Metalloproteinases
keywords: Atherosclerosis; carotid stenosis; cytokines; inflammation