Impaired neutrophil extracellular traps and inflammatory responses in the peritoneal fluid of patients with liver cirrhosis.

Liver cirrhosis (LC) is an inflammatory process associated with impaired functions in adaptive and innate immune responses at both systemic and local levels, also referred as Cirrhosis-Associated Immune Dysfunction. In this study, we evaluated the functionality of neutrophils from ascitic fluid (AF) of patients with hepatic cirrhosis by testing their ability to generate neutrophil extracellular traps (NETs) in vitro. To further determine the activation state of neutrophils, expression of the activation markers CD66b, CD69, and CD80 on these cells was analysed by flow cytometry. The inflammatory environment in AF was assessed by measured concentration of pro- and anti-inflammatory cytokines. Samples were collected from 40 patients with LC, 20 of them with uncomplicated ascites (ASC) and 20 with spontaneous bacterial peritonitis (SBP). Peripheral blood (PB) neutrophils from healthy individuals were used as control (HC). Our results revealed a significant decrease in the release of NETs in neutrophils from the SBP group compared with HC. Low expression of CD69 and CD80 on neutrophils from AF of SBP patients was also observed. Comparisons of inflammatory cytokine levels in AF from the different study groups (SBP and ASC) revealed significant differences. In conclusion, we demonstrate that the development of complications, such as SBP, increases initially the inflammatory status, but chronically results in impaired neutrophil function as demonstrated by the decreased capability of NETs formation. There is also an increase in both pro-inflammatory and anti-inflammatory cytokines, thus predisposing for new episodes of SPB and increasing morbidity and mortality in cirrhotic patients.

Apolipoprotein AI and apolipoprotein E mRNA expression in peripheral white blood cells from patients with orthotopic liver transplantation.

BACKGROUND: Apolipoprotein AI/apolipoprotein E (apo-AI/apo-E) ratio change and its induction in non-hepatic tissues have been reported during liver development, regeneration, and several pathophysiologic states. The clinical implication of such changes is unclear, but these could reflect recovery and/or severity of liver damage. METHODS AND RESULTS: Using RT-PCR we analysed the mRNA expression of apo-AI and apo-E in peripheral white blood cells (PWBC) of patients with different liver diseases who underwent orthotopic liver transplantation (OLT) and compared its expression with the lipid profile and liver function tests. We found that patients showed higher levels of apo-AI mRNA without detection of apo-E mRNA on PWBC at the preoperative day, compared with healthy volunteers (HV). We found an apo-AI/apo-E mRNA ratio of 2.7 during the anhepatic stage, followed by a decrease to 1.3, 0.95, and 0.55 at days 30, 60, and 90, respectively. At the last time point, the apo-AI/apo-E ratio was similar to HV. At day 3 post-OLT, the lowest levels of high-density lipoprotein (HDL)-cholesterol (17 mg/dl; P<0.05) and the highest levels of aspartate aminotransferase, total bilirubin and alkaline phosphatase (77.5 IU/l, 37.9 g/dl, 177.8 IU/l, respectively; P<0.05) were detected. CONCLUSION: These results indicate that changes of HDL-cholesterol and apo-AI/apo-E mRNA ratio could be a good indicator of liver damage and/or hepatic functional recovery post-OLT.