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

Abstract

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.

Neutrophil extracellular traps in patients with liver cirrhosis and hepatocellular carcinoma

“…Our results agree with previous publications such as Blasi et al 29 who have showed that markers of NET are elevated in patients with cirrhosis as compared to healthy controls. However, in contrast to our findings, Agraz-Cibrián et al showed that neutrophils from ascites fluid from cirrhotic patients with spontaneous bacterial peritonitis had reduced ability to form NETs 30 . The same author also showed that neutrophils from peripheral blood of cirrhosis patients also had reduced ability of NET formation upon stimulation in vitro 31 .…”

Single-cell RNA-seq analysis reveals BHLHE40-driven pro-tumour neutrophils with hyperactivated glycolysis in pancreatic tumour microenvironment

“…TAN-1 could be recognised as the ‘pro-tumour subpopulation’, which highly expressed pro-angiogenic factor VEGFA,36 pro-metastatic factor PLAU37 38 and LGALS3, a molecule with multiple functions beneficial for tumour progression, including enhancing proliferation and stemness of malignant cells, promoting angiogenesis, suppressing immune surveillance, promoting M2 macrophage differentiation and contributing to drug resistance39 40 (figure 2D (i)). TAN-2 consisted of an inflammatory subpopulation, which strongly expressed inflammation-associated genes NLRP3 and PDE4B,41 42 and neutrophil activation marker CD6943 44 (figure 2D (ii)). TAN-2 also strongly expressed pro-tumour molecules IL1RN and adrenomedullin (ADM) (figure 2D (ii)), which has been found to support tumour progression in other cancer types 45 46.…”

The value of ascitic neutrophil gelatinase‐associated lipocalin in decompensated liver cirrhosis with spontaneous bacterial peritonitis

“…Once inflammation triggers endotoxins, cytokines, oxidative stress, and apoptosis, returning to baseline is much harder. [20][21][22] Most noteworthy, except for the SBP group, the mortality rate of patients with liver cirrhosis complicated with infection of any type decreased to 31.5%.…”