Skip to main content
Top
Published in: Journal of Translational Medicine 1/2017

Open Access 01-12-2017 | Research

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

Authors: Cornelius Engelmann, Katrin Splith, Sandra Krohn, Adam Herber, Albrecht Boehlig, Stephan Boehm, Johann Pratschke, Thomas Berg, Moritz Schmelzle

Published in: Journal of Translational Medicine | Issue 1/2017

Login to get access

Abstract

Background

Microparticles (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.

Methods

Ascites 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.

Results

MPs 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.

Conclusions

Ascites 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.
Appendix
Available only for authorised users
Literature
1.
go back to reference Boulanger CM. Microparticles, vascular function and hypertension. Curr Opin Nephrol Hypertens. 2010;19:177–80.CrossRefPubMed Boulanger CM. Microparticles, vascular function and hypertension. Curr Opin Nephrol Hypertens. 2010;19:177–80.CrossRefPubMed
2.
go back to reference Dignat-George F, Boulanger CM. The many faces of endothelial microparticles. Arterioscler Thromb Vasc Biol. 2011;31:27–33.CrossRefPubMed Dignat-George F, Boulanger CM. The many faces of endothelial microparticles. Arterioscler Thromb Vasc Biol. 2011;31:27–33.CrossRefPubMed
3.
go back to reference Morel O, Jesel L, Freyssinet JM, Toti F. Cellular mechanisms underlying the formation of circulating microparticles. Arterioscler Thromb Vasc Biol. 2011;31:15–26.CrossRefPubMed Morel O, Jesel L, Freyssinet JM, Toti F. Cellular mechanisms underlying the formation of circulating microparticles. Arterioscler Thromb Vasc Biol. 2011;31:15–26.CrossRefPubMed
4.
go back to reference Feng B, Chen Y, Luo Y, Chen M, Li X, Ni Y. Circulating level of microparticles and their correlation with arterial elasticity and endothelium-dependent dilation in patients with type 2 diabetes mellitus. Atherosclerosis. 2010;208:264–9.CrossRefPubMed Feng B, Chen Y, Luo Y, Chen M, Li X, Ni Y. Circulating level of microparticles and their correlation with arterial elasticity and endothelium-dependent dilation in patients with type 2 diabetes mellitus. Atherosclerosis. 2010;208:264–9.CrossRefPubMed
5.
go back to reference Tramontano AF, Lyubarova R, Tsiakos J, Palaia T, Deleon JR, Ragolia L. Circulating endothelial microparticles in diabetes mellitus. Mediat Inflamm. 2010. doi:10.1155/2010/250476. Tramontano AF, Lyubarova R, Tsiakos J, Palaia T, Deleon JR, Ragolia L. Circulating endothelial microparticles in diabetes mellitus. Mediat Inflamm. 2010. doi:10.​1155/​2010/​250476.
6.
go back to reference Diehl P, Aleker M, Helbing T, et al. Increased platelet, leukocyte and endothelial microparticles predict enhanced coagulation and vascular inflammation in pulmonary hypertension. J Thromb Thrombolysis. 2011;31:173–9.CrossRefPubMed Diehl P, Aleker M, Helbing T, et al. Increased platelet, leukocyte and endothelial microparticles predict enhanced coagulation and vascular inflammation in pulmonary hypertension. J Thromb Thrombolysis. 2011;31:173–9.CrossRefPubMed
7.
go back to reference Pelletier F, Garnache-Ottou F, Angelot F, et al. Increased levels of circulating endothelial-derived microparticles and small-size platelet-derived microparticles in psoriasis. J Investig Dermatol. 2011;131:1573–6.CrossRefPubMed Pelletier F, Garnache-Ottou F, Angelot F, et al. Increased levels of circulating endothelial-derived microparticles and small-size platelet-derived microparticles in psoriasis. J Investig Dermatol. 2011;131:1573–6.CrossRefPubMed
8.
go back to reference Nieslon CT, Østergaard O, Johnsen C, Jacobsen S, Heegaard NH. Distinct features of circulating microparticles and their relationship to clinical manifestations in systemic lupus erythematosus. Arthritis Rheumatol. 2012;63:3067–77. Nieslon CT, Østergaard O, Johnsen C, Jacobsen S, Heegaard NH. Distinct features of circulating microparticles and their relationship to clinical manifestations in systemic lupus erythematosus. Arthritis Rheumatol. 2012;63:3067–77.
9.
go back to reference van Beers EJ, Schaap MC, Berckmans RJ, et al. Circulating erythrocyte-derived microparticles are associated with coagulation activation in sickle cell disease. Haematologica. 2009;94:1513–9.CrossRefPubMedPubMedCentral van Beers EJ, Schaap MC, Berckmans RJ, et al. Circulating erythrocyte-derived microparticles are associated with coagulation activation in sickle cell disease. Haematologica. 2009;94:1513–9.CrossRefPubMedPubMedCentral
10.
go back to reference Omoto S, Nomura S, Shouzu A, et al. Significance of platelet derived microparticles and activated platelets in diabetic nephropathy. Nephron. 1999;81:271–7.CrossRefPubMed Omoto S, Nomura S, Shouzu A, et al. Significance of platelet derived microparticles and activated platelets in diabetic nephropathy. Nephron. 1999;81:271–7.CrossRefPubMed
11.
go back to reference Biasucci LM, Porto I, Vito LD, et al. Differences in microparticle release in patients with acute coronary syndrome and stable angina. Circ J. 2012;76:2174–82.CrossRefPubMed Biasucci LM, Porto I, Vito LD, et al. Differences in microparticle release in patients with acute coronary syndrome and stable angina. Circ J. 2012;76:2174–82.CrossRefPubMed
12.
go back to reference Nomura S, Inami N, Shouzu A, et al. Effects of pitavastatin, eicosapentaenoic acid and combined therapy on platelet-derived microparticles and adiponectin in hyperlipidemic, diabetic patients. Platelets. 2009;20:16–22.CrossRefPubMed Nomura S, Inami N, Shouzu A, et al. Effects of pitavastatin, eicosapentaenoic acid and combined therapy on platelet-derived microparticles and adiponectin in hyperlipidemic, diabetic patients. Platelets. 2009;20:16–22.CrossRefPubMed
13.
go back to reference Brodsky SV, Zhang F, Nasjletti A, Goligorsky MS. Endothelium-derived microparticles impair endothelial function in vitro. Am J Physiol Heart Circ Physiol. 2004;286:H1910–5.CrossRefPubMed Brodsky SV, Zhang F, Nasjletti A, Goligorsky MS. Endothelium-derived microparticles impair endothelial function in vitro. Am J Physiol Heart Circ Physiol. 2004;286:H1910–5.CrossRefPubMed
15.
go back to reference Kornek M, Popov Y, Libermann TA, Afdhal NH, Schuppan D. Human T cell microparticles circulate in blood of hepatitis patients and induce fibrolytic activation of hepatic stellate cells. Hepatology. 2011;53:230.CrossRefPubMed Kornek M, Popov Y, Libermann TA, Afdhal NH, Schuppan D. Human T cell microparticles circulate in blood of hepatitis patients and induce fibrolytic activation of hepatic stellate cells. Hepatology. 2011;53:230.CrossRefPubMed
16.
go back to reference Kornek M, Lynch M, Metha SH, et al. Circulating microparticles as disease-specific biomarker of severity of inflammation in patients with hepatitis C or nonalcoholic steatohepatitis. Gastroenterology. 2012;143:448–58.CrossRefPubMedPubMedCentral Kornek M, Lynch M, Metha SH, et al. Circulating microparticles as disease-specific biomarker of severity of inflammation in patients with hepatitis C or nonalcoholic steatohepatitis. Gastroenterology. 2012;143:448–58.CrossRefPubMedPubMedCentral
17.
go back to reference Schmelzle M, Splith K, Andersen LW, et al. Increased plasma levels of microparticles expressing CD39 and CD133 in acute liver injury. Transplantation. 2013;95:63–9.CrossRefPubMedPubMedCentral Schmelzle M, Splith K, Andersen LW, et al. Increased plasma levels of microparticles expressing CD39 and CD133 in acute liver injury. Transplantation. 2013;95:63–9.CrossRefPubMedPubMedCentral
18.
go back to reference Berckmans RJ, Nieuwland R, Tak PP, et al. Cell-derived microparticles in synovial fluid from inflamed arthritic joints support coagulation exclusively via factor VII-dependent mechanisms. Arthritis Rheumatol. 2002;46:2857–66.CrossRef Berckmans RJ, Nieuwland R, Tak PP, et al. Cell-derived microparticles in synovial fluid from inflamed arthritic joints support coagulation exclusively via factor VII-dependent mechanisms. Arthritis Rheumatol. 2002;46:2857–66.CrossRef
19.
go back to reference Gutwein P, Stoeck A, Riedle S, et al. Cleavage of L1 in exosomes and apoptotic membrane vesicles released from ovarian carcinoma cells. Clin Cancer Res. 2005;11:2492–501.CrossRefPubMed Gutwein P, Stoeck A, Riedle S, et al. Cleavage of L1 in exosomes and apoptotic membrane vesicles released from ovarian carcinoma cells. Clin Cancer Res. 2005;11:2492–501.CrossRefPubMed
20.
go back to reference Pascual M, Steiger G, Sadallah S, et al. Identification of membrane-bound CR1 (CD35) in human urine: evidence for its release by glomerular podocytes. J Exp Med. 1994;179:889–99.CrossRefPubMed Pascual M, Steiger G, Sadallah S, et al. Identification of membrane-bound CR1 (CD35) in human urine: evidence for its release by glomerular podocytes. J Exp Med. 1994;179:889–99.CrossRefPubMed
21.
go back to reference Mrvar-Brecko A, Sustar V, Jansa V, et al. Isolated microvesicles from peripheral blood and body fluids as observed by scanning electron microscope. Blood Cells Mol Dis. 2010;44:307–12.CrossRefPubMed Mrvar-Brecko A, Sustar V, Jansa V, et al. Isolated microvesicles from peripheral blood and body fluids as observed by scanning electron microscope. Blood Cells Mol Dis. 2010;44:307–12.CrossRefPubMed
22.
go back to reference Press JZ, Reyes M, Pitteri SJ, et al. Microparticles from ovarian carcinomas are shed into ascites and promote cell migration. Int J Gynecol Cancer. 2012;22:546–52.CrossRefPubMed Press JZ, Reyes M, Pitteri SJ, et al. Microparticles from ovarian carcinomas are shed into ascites and promote cell migration. Int J Gynecol Cancer. 2012;22:546–52.CrossRefPubMed
23.
go back to reference Wong CL, Holroyed-Leduc J, Thorpe KE, Straus SE. Does this patient have bacterial peritonitis or portal hypertension? How do I perform a paracentesis and analyze the results? JAMA. 2008;299:1166–78.CrossRefPubMed Wong CL, Holroyed-Leduc J, Thorpe KE, Straus SE. Does this patient have bacterial peritonitis or portal hypertension? How do I perform a paracentesis and analyze the results? JAMA. 2008;299:1166–78.CrossRefPubMed
24.
go back to reference European Association for the Study of the Liver. EASL clinical practice guidelines on the management of ascites, spontaneous bacterial peritonitis, and hepatorenal syndrome in cirrhosis. J Hepatol. 2010;53(3):397–417.CrossRef European Association for the Study of the Liver. EASL clinical practice guidelines on the management of ascites, spontaneous bacterial peritonitis, and hepatorenal syndrome in cirrhosis. J Hepatol. 2010;53(3):397–417.CrossRef
25.
go back to reference Cloutier N, Tan S, Boudreau LH, et al. The exposure of autoantigens by microparticles underlies the formation of potent inflammatory components: the microparticle-associated immune complexes. EMBO Mol Med. 2013;5:235–49.CrossRefPubMed Cloutier N, Tan S, Boudreau LH, et al. The exposure of autoantigens by microparticles underlies the formation of potent inflammatory components: the microparticle-associated immune complexes. EMBO Mol Med. 2013;5:235–49.CrossRefPubMed
26.
go back to reference Arroyo V, Moreau R, Jalan R, Gines P. Acute-on-chronic liver failure: a new syndrome that will re-classify cirrhosis. J Hepatol. 2015;62:S131–43.CrossRefPubMed Arroyo V, Moreau R, Jalan R, Gines P. Acute-on-chronic liver failure: a new syndrome that will re-classify cirrhosis. J Hepatol. 2015;62:S131–43.CrossRefPubMed
27.
go back to reference Claria J, Strauber RE, Coenraad MJ, et al. Systemic inflammation in decompensated cirrhosis: characterization and role in acute-on-chronic liver failure. Hepatology. 2016;64(4):1249–64.CrossRefPubMed Claria J, Strauber RE, Coenraad MJ, et al. Systemic inflammation in decompensated cirrhosis: characterization and role in acute-on-chronic liver failure. Hepatology. 2016;64(4):1249–64.CrossRefPubMed
28.
go back to reference Yun CH, Holroyd-Leduc J, Thorpe KE, Straus SE. Does this patient have bacterial peritonitis or portal hypertension? How do I perform a paracentesis and analyze the results? J Clin Neurol. 2010;6:89–98.CrossRefPubMedPubMedCentral Yun CH, Holroyd-Leduc J, Thorpe KE, Straus SE. Does this patient have bacterial peritonitis or portal hypertension? How do I perform a paracentesis and analyze the results? J Clin Neurol. 2010;6:89–98.CrossRefPubMedPubMedCentral
29.
go back to reference Esposito K, Ciotola M, Schisano B, et al. Endothelial microparticles correlate with endothelial dysfunction in obese women. J Clin Endocrinol Metab. 2006;91:3676–9.CrossRefPubMed Esposito K, Ciotola M, Schisano B, et al. Endothelial microparticles correlate with endothelial dysfunction in obese women. J Clin Endocrinol Metab. 2006;91:3676–9.CrossRefPubMed
30.
go back to reference Amabile N, Guérin AP, Tedgui A, Boulanger CM, London GM. Predictive value of circulating endothelial microparticles for cardiovascular mortality in end-stage renal failure: a pilot study. Nephrol Dial Transplant. 2012;27:1873–80.CrossRefPubMed Amabile N, Guérin AP, Tedgui A, Boulanger CM, London GM. Predictive value of circulating endothelial microparticles for cardiovascular mortality in end-stage renal failure: a pilot study. Nephrol Dial Transplant. 2012;27:1873–80.CrossRefPubMed
31.
go back to reference Park MS, Owen BA, Ballinger BA, et al. Quantification of hypercoagulable state after blunt trauma: microparticle and thrombin generation are increased relative to injury severity, while standard markers are not. Surgery. 2012;151:831–6.CrossRefPubMedPubMedCentral Park MS, Owen BA, Ballinger BA, et al. Quantification of hypercoagulable state after blunt trauma: microparticle and thrombin generation are increased relative to injury severity, while standard markers are not. Surgery. 2012;151:831–6.CrossRefPubMedPubMedCentral
32.
go back to reference Knijff-Dutmer EA, Koerts J, Nieuwland R, Kalsbeek-Batenburg EM, van de Laar MA. Elevated levels of platelet microparticles are associated with disease activity in rheumatoid arthritis. Arthritis Rheumatol. 2002;46:1498–503.CrossRef Knijff-Dutmer EA, Koerts J, Nieuwland R, Kalsbeek-Batenburg EM, van de Laar MA. Elevated levels of platelet microparticles are associated with disease activity in rheumatoid arthritis. Arthritis Rheumatol. 2002;46:1498–503.CrossRef
33.
go back to reference Umekita K, Hidaka T, Ueno S, et al. Leukocytapheresis (LCAP) decreases the level of platelet-derived microparticles (MPs) and increases the level of granulocytes-derived MPs: a possible connection with the effect of LCAP on rheumatoid arthritis. Mod Rheumatol. 2009;19:265–72.CrossRefPubMed Umekita K, Hidaka T, Ueno S, et al. Leukocytapheresis (LCAP) decreases the level of platelet-derived microparticles (MPs) and increases the level of granulocytes-derived MPs: a possible connection with the effect of LCAP on rheumatoid arthritis. Mod Rheumatol. 2009;19:265–72.CrossRefPubMed
34.
go back to reference Tamagawa-Mineoka J, Katoh N, Kishimoto S. Platelet activation in patients with psoriasis: increased plasma levels of platelet-derived microparticles and soluble P-selectin. Am Acad Dermatol. 2010;62:621–6.CrossRef Tamagawa-Mineoka J, Katoh N, Kishimoto S. Platelet activation in patients with psoriasis: increased plasma levels of platelet-derived microparticles and soluble P-selectin. Am Acad Dermatol. 2010;62:621–6.CrossRef
36.
go back to reference Simonetto DA, Yang HY, Yin M, et al. Chronic passive venous congestion drives hepatic fibrogenesis via sinusoidal thrombosis and mechanical forces. Hepatology. 2015;61:648–59.CrossRefPubMedPubMedCentral Simonetto DA, Yang HY, Yin M, et al. Chronic passive venous congestion drives hepatic fibrogenesis via sinusoidal thrombosis and mechanical forces. Hepatology. 2015;61:648–59.CrossRefPubMedPubMedCentral
37.
go back to reference Rautou PE, Bresson J, Sainte-Marie Y, et al. Abnormal plasma microparticles impair vasoconstrictor responses in patients with cirrhosis. Gastroenterology. 2012;143:166–76.CrossRefPubMed Rautou PE, Bresson J, Sainte-Marie Y, et al. Abnormal plasma microparticles impair vasoconstrictor responses in patients with cirrhosis. Gastroenterology. 2012;143:166–76.CrossRefPubMed
38.
go back to reference Ogasawara F, Fusegawa H, Haruki Y, Shiraishi K, Watanabe N, Matsuzaki S. Platelet activation in patients with alcoholic liver disease. Tokai J Exp Clin Med. 2005;30:41–8.PubMed Ogasawara F, Fusegawa H, Haruki Y, Shiraishi K, Watanabe N, Matsuzaki S. Platelet activation in patients with alcoholic liver disease. Tokai J Exp Clin Med. 2005;30:41–8.PubMed
Metadata
Title
Absolute quantification of microparticles by flow cytometry in ascites of patients with decompensated cirrhosis: a cohort study
Authors
Cornelius Engelmann
Katrin Splith
Sandra Krohn
Adam Herber
Albrecht Boehlig
Stephan Boehm
Johann Pratschke
Thomas Berg
Moritz Schmelzle
Publication date
01-12-2017
Publisher
BioMed Central
Published in
Journal of Translational Medicine / Issue 1/2017
Electronic ISSN: 1479-5876
DOI
https://doi.org/10.1186/s12967-017-1288-3

Other articles of this Issue 1/2017

Journal of Translational Medicine 1/2017 Go to the issue