Circulating Neutrophils of Nonalcoholic Steatohepatitis Patients Show an Activated Phenotype and Suppress T Lymphocytes Activity
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 CD16HighCD62LHighCD11bHigh 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 CD16HighCD62LDimCD11bHigh 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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