Absolute quantification of microparticles by flow cytometry in ascites of patients with decompensated cirrhosis: a cohort study

Abstract

BackgroundMicroparticles (MPs) are small (<1 μm) cell membrane-derived vesicles that are formed in response to cellular activation or early stages of apoptosis. Increased plasma MP levels have been associated with liver disease severity. Here we investigated the clinical impact of ascites MPs in patients with decompensated liver cirrhosis.MethodsAscites and blood samples of 163 patients with cirrhosis (ascites n = 163, blood n = 31) were collected between February 2011 and December 2012. MPs were obtained from ascites and from blood by two-step ultracentrifugation and quantified by flow cytometry. Quantitative absolute MP levels were correlated with clinical and laboratory baseline parameters as well as patient outcomes. Ascites microparticles were stained with antibodies against CD66b (neutrophils) and CD3 (lymphocytes) in a subgroup of 60 matched patients.ResultsMPs were detected in all ascites and blood samples. Absolute ascites MP levels correlated with blood levels (r = 0.444, p = 0.011). Low ascites MP levels (<488.4 MP/μL) were associated with a poor 30-day survival probability (<488.4 MP/μL 71.1% vs. >488.4 MP/μL 94.7%, log rank p = 0.001) and such patients had a higher relative amount of ascites microparticles derived from neutrophils and lymphocytes. Low levels of ascites MPs, high MELD score and antibiotic treatment were independent risk factors for death within 30 days.ConclusionsAscites MP levels predict short-term survival along with the liver function in patients with decompensated cirrhosis. Further studies which evaluate ascites MPs as disease specific biomarker with a validation cohort and which investigate its underlying mechanisms are needed. Neutrophils and lymphocytes contributed more frequently to the release of microparticles in patients with low ascites levels, possibly indicating an immune activation in this cohort.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-017-1288-3) contains supplementary material, which is available to authorized users.

Extracellular Vesicles in Liver Diseases: Diagnostic, Prognostic, and Therapeutic Application

“…13 On the other hand, patients with low levels of EVs in ascites (< 488.4 MV/µL, n ¼ 101) had a lower 30-day survival rate of 71.7% when compared with 94.7% in patients with high levels of EVs in ascites (> 488.4 EV/µL, n ¼ 62, p ¼ 0.0001). 14 In experimental models of nonalcoholic steatohepatitis (NASH) or ASH, hepatocyte-derived EVs released from damaged/stressed hepatocytes contributed to the progression of liver disease through activation of liver endothelial cells, HSCs, and hepatic macrophages. 26,[40][41][42][43] In terms of human subjects, the pioneering study was published just a few years ago by Kornek et al For the first time, they suggested correlations between the circulating abundance of leukocyte-derived EVs and disease severity, as determined by liver transaminase levels, biopsy grade, and NAFLD activity score.…”

“…In a similar fashion, different stages of disease severity equal different loads of EVs; hence, they additionally are potential prognostic biomarkers. 13,14 It would be reasonable to speculate that once the parental cells are removed, this will lead to a clearance of EVs, 15 highlighting another role of EVs: a tool for monitoring treatments and treatment outcomes. Nevertheless, the ability of selectively manipulating EV content opens new frontiers in the EV world: the use of EVs in disease therapeutics.…”

Extracellular vesicles: Future diagnostic and therapeutic tools for liver disease and regeneration

“…They reported that leucocyte‐derived CD14‐ and CD16‐positive EV counts were correlated with the severity of fibrosis . A recent study characterized the surface molecules of EV in ascites and reported that a low EV count with relatively high CD66b‐ and CD3‐positive EV levels correlated with a poor 30‐day survival . Furthermore, several studies point out that the miRNA profile of plasma‐derived EV could be used as a biomarker of fibrosis in patients.…”

Changes in plasma circulating microvesicles in patients with HCV‐related cirrhosis after treatment with direct‐acting antivirals

“…Particularly, microvesicles (MVs), also referred to as ‘microparticles’, are 100‐1000 nm diameter‐EVs released via outward budding of the cell plasma membrane both constitutively and in response to various stimulations. As MVs act as important messengers in intercellular communication, they are involved in various pathomechanistic processes and increasing MV levels generally express disease activity and progression . Specifically, MVs have been shown to play a role as procoagulant and pro‐inflammatory mediators.…”

“…As MVs act as important messengers in intercellular communication, they are involved in various pathomechanistic processes and increasing MV levels generally express disease activity and progression. 1,[4][5][6] Specifically, MVs have been shown to play a role as procoagulant and pro-inflammatory mediators.…”