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NOVEL CD14HICD16HI SUBSET OF PERITONEAL MACROPHAGE FROM CIRRHOTIC PATIENTS IS ASSOCIATED TO AN INCREASED RESPONSE TO LPS
Autores:
ANTONIO JOSÉ RUIZ ALCARAZ
,
ANA TAPIA ABELLÁN
, MARÍA DOLORES FERNÁNDEZ FERNÁNDEZ, MARÍA TRISTÁN MANZANO,
TRINIDAD HERNÁNDEZ CASELLES
, EDUARDO SÁNCHEZ VELASCO, MANUEL MIRAS LÓPEZ,
MARIA PILAR GARCÍA PEÑARRUBIA
,
MARÍA CONCEPCIÓN MARTÍNEZ-ESPARZA ALVARGONZÁLEZ
,
Grupos de investigación:
[GI/IMIB/C063/2011] Inmunidad innata en la salud y la enfermedad
[GI/IMIB/C020/2011] Investigación en Pediatría
[GI/IMIB/C003/2011] Cirugía digestiva, endocrina y trasplante de órganos abdominales
Comunicación:
Antecedentes:
A better knowledge of the phenotype, the activation status and the capacity of response of resident macrophages present in the ascites fluid (AF) of cirrhotic patients would be useful to identify novel targets for pharmaceutical intervention to prevent hepatic damage. 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.
Métodos:
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.
Resultados:
A novel CD14highCD16high 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-, and lower levels of IL-1 and IL-12 than control blood M-DM.
Conclusiones:
In conclusion, a new subpopulation of CD14highCD16high peritoneal M-DM has been identified in ascites of cirrhotic patients, which is very sensitive to LPS stimulation.
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