Circulating Neutrophils of Nonalcoholic Steatohepatitis Patients Show an Activated Phenotype and Suppress T Lymphocytes Activity

Figure 4

PMN ROS intracellular production and viability. (a) PMNs obtained from healthy donors, NAFL, and NASH patients after 24 h of coculture, with (PMNs+PBMCs) or without PBMCs (PMNs), were stained with H2DCF-DA probe for intracellular production of ROS and (b) Annexin-V/Live Dead for vitality. (a) Left panel: histogram of H2DCFDA (ROS) MFI of CD16^HighCD62L^HighCD11b^High cell subset obtained from healthy donors (), NAFL (), and NASH (). Right panel: representative histogram of flow cytometry analysis with an overlay showing H2DCFDA (ROS) MFI in gated CD16^HighCD62L^DimCD11b^High of healthy donors (dashed line), NAFL (light gray), and NASH patients (dark gray), after 24 h of coculture with PBMCs. FMO control is represented by gray histogram and LPS positive control by a gray line. (b) Left panel: representative plots of PMNs cell death of healthy donors, NAFL, and NASH patients, after 24 h, with (PMNS+PBMCs) or without PBMCs (PMNs). Right panel: percentage of live (Annexin-V, Live Dead negative) and early apoptotic (Annexin-V positive, Live Dead negative) of PMNs obtained from healthy donors (), NAFL (), and NASH patients ().In all histograms reported, for each subgroup, the distribution of each subject/experiment is showed and results are expressed as . Each dot represents one subject/experiment. Each subject has been studied in an independent experiment. In all experiments, statistics were performed by Mann-Whitney unpaired test, 2-tailed. value, , , , , ns: nonsignificant.

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