title: Characterisation of leukocytes in a human skin blister model of acute inflammation and resolution.
abstract: There is an increasing need to understand the leukocytes and soluble mediators that drive acute inflammation and bring about its resolution in humans. We therefore carried out an extensive characterisation of the cantharidin skin blister model in healthy male volunteers. A novel fluorescence staining protocol was designed and implemented, which facilitated the identification of cell populations by flow cytometry. We observed that at the onset phase, 24 h after blister formation, the predominant cells were CD16hi/CD66b+ PMNs followed by HLA-DR+/CD14+ monocytes/macrophages, CD11c+ and CD141+ dendritic cells as well as Siglec-8+ eosinophils. CD3+ T cells, CD19+ B cells and CD56+ NK cells were also present, but in comparatively fewer numbers. During resolution, 72 h following blister induction, numbers of PMNs declined whilst the numbers of monocyte/macrophages remain unchanged, though they upregulated expression of CD16 and CD163. In contrast, the overall numbers of dendritic cells and Siglec-8+ eosinophils increased. Post hoc analysis of these data revealed that of the inflammatory cytokines measured, TNF-α but not IL-1β or IL-8 correlated with increased PMN numbers at the onset. Volunteers with the greatest PMN infiltration at onset displayed the fastest clearance rates for these cells at resolution. Collectively, these data provide insight into the cells that occupy acute resolving blister in humans, the soluble mediators that may control their influx as well as the phenotype of mononuclear phagocytes that predominate the resolution phase. Further use of this model will improve our understanding of the evolution and resolution of inflammation in humans, how defects in these over-lapping pathways may contribute to the variability in disease longevity/chronicity, and lends itself to the screen of putative anti-inflammatory or pro-resolution therapies.
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: William
authors.initials: W
authors.lastname: Jenner
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: Madhur
authors.initials: M
authors.lastname: Motwani
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: Kristin
authors.initials: K
authors.lastname: Veighey
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: Justine
authors.initials: J
authors.lastname: Newson
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: Tatsiana
authors.initials: T
authors.lastname: Audzevich
authors.affiliation: Manchester Pharmacy School, Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Nicolaou
authors.affiliation: Manchester Pharmacy School, Faculty of Medical and Human Sciences, University of Manchester, Manchester, United Kingdom.
authors.forename: Sharon
authors.initials: S
authors.lastname: Murphy
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: Raymond
authors.initials: R
authors.lastname: Macallister
authors.affiliation: Centre for Clinical Pharmacology and Therapeutics, Division of Medicine, University College London, London, United Kingdom.
authors.forename: Derek W
authors.initials: DW
authors.lastname: Gilroy
mesh_terms: D000293:Adolescent; D000328:Adult; D018941:Antigens, CD19 / Q000276:immunology / Q000378:metabolism; D001402:B-Lymphocytes / Q000276:immunology; D001768:Blister / Q000276:immunology* / Q000378:metabolism; D017252:CD3 Complex / Q000276:immunology / Q000378:metabolism; D019002:CD56 Antigen / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D004804:Eosinophils / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007249:Inflammation / Q000276:immunology* / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology / Q000378:metabolism; D007958:Leukocyte Count; D007962:Leukocytes / Q000276:immunology* / Q000378:metabolism; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000276:immunology / Q000378:metabolism; D020556:Neutrophil Infiltration / Q000276:immunology; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism; D012867:Skin / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D055815:Young Adult
chemical_list: D018941:Antigens, CD19; D017252:CD3 Complex; D019002:CD56 Antigen; D016207:Cytokines