title: A novel CD14(high) CD16(high) subset of peritoneal macrophages from cirrhotic patients is associated to an increased response to LPS.
abstract: The aim of this study was to characterize monocyte-derived macrophages (M-DM) from blood and ascites of cirrhotic patients comparatively with those obtained from blood of healthy controls. The phenotypic profile based on CD14/CD16 expression was analyzed by flow cytometry. Cells were isolated and stimulated in vitro with LPS and heat killed Candida albicans. Phosphorylation of ERK, c-Jun, p38 MAPK, and PKB/Akt was analyzed by Western blotting. A novel CD14(high)CD16(high) M-DM subpopulation is present in ascites (∼33%). The CD14(++)CD16(+) intermediate subset is increased in the blood of cirrhotic patients (∼from 4% to 11%) and is predominant in ascites (49%), while the classical CD14(++)CD16(-) subpopulation is notably reduced in ascites (18%). Basal hyperactivation of ERK and JNK/c-Jun pathways observed in ascites M-DM correlates with CD14/CD16 high expressing subsets, while PI3K/PKB does it with the CD16 low expressing cells. In vitro LPS treatment highly increases ERK1/2, PKB/Akt and c-Jun phosphorylation, while that of p38 MAPK is decreased in M-DM from ascites compared to control blood M-DM. Stimulation of healthy blood M-DM with LPS and C. albicans induced higher phosphorylation levels of p38 than those from ascites. Regarding cytokines secretion, in vitro activated M-DM from ascites of cirrhotic patients produced significantly higher amounts of IL-6, IL-10 and TNF-α, and lower levels of IL-1β and IL-12 than control blood M-DM. In conclusion, a new subpopulation of CD14(high)CD16(high) peritoneal M-DM has been identified in ascites of cirrhotic patients, which is very sensitive to LPS stimulation.
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain.
authors.forename: Antonio José
authors.initials: AJ
authors.lastname: Ruiz-Alcaraz
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Tapia-Abellán
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain.
authors.forename: María Dolores
authors.initials: MD
authors.lastname: Fernández-Fernández
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Tristán-Manzano
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain.
authors.forename: Trinidad
authors.initials: T
authors.lastname: Hernández-Caselles
authors.affiliation: Unidad de Trasplante Hepático, Servicio de Aparato Digestivo, Hospital Universitario Virgen de la Arrixaca, IMIB, Murcia, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Sánchez-Velasco
authors.affiliation: Unidad de Trasplante Hepático, Servicio de Aparato Digestivo, Hospital Universitario Virgen de la Arrixaca, IMIB, Murcia, Spain.
authors.forename: Manuel
authors.initials: M
authors.lastname: Miras-López
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain. Electronic address: maria@um.es.
authors.forename: María
authors.initials: M
authors.lastname: Martínez-Esparza
authors.affiliation: Departamento de Bioquímica, Biología Molecular (B) e Inmunología, Facultad de Medicina, IMIB and Regional Campus of International Excellence "Campus Mare Nostrum", Universidad de Murcia, 30100 Murcia, Spain. Electronic address: pigarcia@um.es.
authors.forename: Pilar
authors.initials: P
authors.lastname: García-Peñarrubia
mesh_terms: D000328:Adult; D000368:Aged; D001201:Ascites / Q000276:immunology; D002176:Candida albicans / Q000276:immunology; D016207:Cytokines / Q000378:metabolism; D005260:Female; D006801:Humans; D018950:Lipopolysaccharide Receptors / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology; D008103:Liver Cirrhosis / Q000276:immunology*; D017737:Macrophages, Peritoneal / Q000276:immunology*; D008297:Male; D008875:Middle Aged; D017452:Receptors, IgG / Q000276:immunology; D015398:Signal Transduction; D055815:Young Adult
chemical_list: D016207:Cytokines; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D017452:Receptors, IgG
keywords: Ascites; Cell signaling; Cirrhosis; Cytokines; Inflammation; Monocyte subsets