The immunological response of innate cells is altered regardless the severity of Non-alcoholic fatty liver disease (NAFLD) progression in children and adults.

INZAUGARAT ME, CERVINI G, VODANOVICH F, BILLORDO LA, CASAVALLE P, VEDIRE C, GALOPPO C, GONZALEZ BALLERGA E, DARUICH J, SORDA JA, CHERÑAVSKY AC.

Amsterdam

Congreso; 48th Annual Meeting of the European Association for the Study of the Liver (EASL).; 2013

EASLD

Background and aims: Obesity increases the risk for metabolic diseases. NAFLD ranges from simple steatosis (SS) to non-alcoholic steatohepatitis (NASH). Reactive oxygen species (ROS) and cytokines production by Kuppfer Cells (KC), monocytes (Mo) and T cells are associated w ith liver damage. We aimed to evaluate alterations of the immunological response during NAFLD progression. Patients and methods: Peripheral blood mononuclear cells (PBMC) w ere obtained from adults (AD) w ith NAFLD (NAFLDAD, n=60), healthy controls (Co AD, n=30), obese children (OB CH)w ith/w ithout NASH (n=70) and Co CH (n=30), and liver biopsies from NAFLDAD. To evaluate ROS production, PMBC or chemically / mechanically obtained liver cell suspensions w ere incubated w ith dichlorofluorescein-diacetate (5 μM), stimulated w ith leptin [Lep (10nM)], palmitic and linoleic acids [PA (500 μM)], LA(200μM)], stained w ith anti-CD14 (Mo) or anti-CD14 /-CD11b mAbs(KC) and analyzed by flow cytometry. A stimulation index (SI) results from the mean fluorescence intensity (MFI) in stimulated / non-stimulated cells. Intracellular TNFa w as evaluated in Lep- stimulated PBMC and a fold of increase index (FI) calculated for CD14+ TNFa+ cells. Production of IFN-γ after PMA stimulation (25 ng/ml) w as evaluated in CD4+ or CD8+cells. Mann-Whitney (p), Wilcoxon matched paired (pp) and chi- square association tests w ere conveniently used. Results: LA induced higher production of ROS in Mo from NAFLDAD (p=0.032) and OB CH (p=0.015)vs. controls. LA, PA and Lep stimulated the oxidative burst in KC from NAFLDAD (pp=0.001, 0.010 and 0.045 respectively). Leptin induced a higher production of TNFa in NAFLDAD (pc=0.009) and OB CH (p=0.009) vs. controls. IFNγ- producing CD4 and CD8 subpopulations w ere larger in NAFLDAD (p=0.007 and 0.08) and OB CH (p=0.07 and 0.005) vs. controls. Significant associations w ere found betw een NASHAD and NASH CH w ith CD4+ IFN γ + (p=0.029 and 0.013) and CD8+ IFN γ + (p=0.036 and 0.039) but not betw een ROS and TNFa production in Mo/CK w ith NASHAD or SSAD, or OB CH w ith/w ithout NASH by chi-square association tests. Conclusions: Given the immunological alterations found in innate cells, SS should not be considered a benign condition. Furthermore, immunological alterations may be involved in NAFLD progression.