Abnormal neutrophil traps and impaired efferocytosis contribute to liver injury and sepsis severity after binge alcohol use

Terence Ndonyi Bukong, Yeonhee Cho, Arvin Iracheta-Vellve, Banishree Saha, Patrick Lowe, Adeyinka Adejumo, Istvan Furi, Aditya Ambade, Benedek Gyongyosi, Donna Catalano, Karen Kodys, G. Szabó

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background & Aims: Neutrophil extracellular traps (NETs) are an important strategy utilized by neutrophils to immobilize and kill invading microorganisms. Herein, we studied NET formation and the process of neutrophil cell death (NETosis), as well as the clearance of NETs by macrophages (MΦ) (efferocytosis) in acute sepsis following binge drinking. Methods: Healthy volunteers consumed 2 ml of vodka/kg body weight, before blood endotoxin and 16 s rDNA were measured. Peripheral neutrophils were isolated and exposed to alcohol followed by phorbol 12-myristate 13-acetate (PMA) stimulation. Mice were treated with three alcohol binges and intraperitoneal lipopolysaccharide (LPS) to assess the dynamics of NET formation and efferocytosis. In vivo, anti-Ly6G antibody (IA8) was used for neutrophil depletion. Results: Inducers of NETs (endotoxin and bacterial DNA) significantly increased in the circulation after binge alcohol drinking in humans. Ex vivo, alcohol alone increased NET formation but attenuated NET formation upon PMA stimulation. Binge alcohol in mice resulted in a biphasic response to LPS. Initially, binge alcohol reduced LPS-induced NET formation and resulted in a diffuse distribution of neutrophils in the liver compared to alcohol-naïve mice. Moreover, indicators of NET formation including citrullinated histone H3, neutrophil elastase, and neutrophil myeloperoxidase were decreased at an early time point after LPS challenge in mice receiving binge alcohol, suggesting decreased NET formation. However, in the efferocytosis phase (15 h after LPS) citrullinated histone-H3 was increased in the liver in alcohol binge mice, suggesting decreased clearance of NETs. In vitro alcohol treatment reduced efferocytosis and phagocytosis of NETotic neutrophils and promoted expression of CD206 on MΦ. Finally, depletion of neutrophils prior to binge alcohol ameliorated LPS-induced systemic inflammation and liver injury in mice. Conclusions: Dysfunctional NETosis and efferocytosis following binge drinking exacerbate liver injury associated with sepsis. Lay summary: Disease severity in alcoholic liver disease (ALD) is associated with a significant presence of neutrophils (a type of immune cell) in the liver. It remains unknown how alcohol affects the capacity of neutrophils to control infection, a major hallmark of ALD. We found that binge alcohol drinking impaired important strategies used by neutrophils to contain and resolve infection, resulting in increased liver injury during ALD.

Original languageEnglish
JournalJournal of Hepatology
DOIs
Publication statusAccepted/In press - Jan 1 2018

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Sepsis
Neutrophils
Alcohols
Liver
Wounds and Injuries
Binge Drinking
Lipopolysaccharides
Alcoholic Liver Diseases
Endotoxins
Alcohol Drinking
Histones
Acetates
Extracellular Traps
Bacterial DNA
Leukocyte Elastase
Infection Control
Ribosomal DNA
Phagocytosis
Peroxidase
Anti-Idiotypic Antibodies

Keywords

  • Alcoholic hepatitis
  • Binge drinking
  • Neutrophil depletion
  • Neutrophil elastase
  • Sepsis

ASJC Scopus subject areas

  • Hepatology

Cite this

Abnormal neutrophil traps and impaired efferocytosis contribute to liver injury and sepsis severity after binge alcohol use. / Bukong, Terence Ndonyi; Cho, Yeonhee; Iracheta-Vellve, Arvin; Saha, Banishree; Lowe, Patrick; Adejumo, Adeyinka; Furi, Istvan; Ambade, Aditya; Gyongyosi, Benedek; Catalano, Donna; Kodys, Karen; Szabó, G.

In: Journal of Hepatology, 01.01.2018.

Research output: Contribution to journalArticle

Bukong, TN, Cho, Y, Iracheta-Vellve, A, Saha, B, Lowe, P, Adejumo, A, Furi, I, Ambade, A, Gyongyosi, B, Catalano, D, Kodys, K & Szabó, G 2018, 'Abnormal neutrophil traps and impaired efferocytosis contribute to liver injury and sepsis severity after binge alcohol use', Journal of Hepatology. https://doi.org/10.1016/j.jhep.2018.07.005
Bukong, Terence Ndonyi ; Cho, Yeonhee ; Iracheta-Vellve, Arvin ; Saha, Banishree ; Lowe, Patrick ; Adejumo, Adeyinka ; Furi, Istvan ; Ambade, Aditya ; Gyongyosi, Benedek ; Catalano, Donna ; Kodys, Karen ; Szabó, G. / Abnormal neutrophil traps and impaired efferocytosis contribute to liver injury and sepsis severity after binge alcohol use. In: Journal of Hepatology. 2018.
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abstract = "Background & Aims: Neutrophil extracellular traps (NETs) are an important strategy utilized by neutrophils to immobilize and kill invading microorganisms. Herein, we studied NET formation and the process of neutrophil cell death (NETosis), as well as the clearance of NETs by macrophages (MΦ) (efferocytosis) in acute sepsis following binge drinking. Methods: Healthy volunteers consumed 2 ml of vodka/kg body weight, before blood endotoxin and 16 s rDNA were measured. Peripheral neutrophils were isolated and exposed to alcohol followed by phorbol 12-myristate 13-acetate (PMA) stimulation. Mice were treated with three alcohol binges and intraperitoneal lipopolysaccharide (LPS) to assess the dynamics of NET formation and efferocytosis. In vivo, anti-Ly6G antibody (IA8) was used for neutrophil depletion. Results: Inducers of NETs (endotoxin and bacterial DNA) significantly increased in the circulation after binge alcohol drinking in humans. Ex vivo, alcohol alone increased NET formation but attenuated NET formation upon PMA stimulation. Binge alcohol in mice resulted in a biphasic response to LPS. Initially, binge alcohol reduced LPS-induced NET formation and resulted in a diffuse distribution of neutrophils in the liver compared to alcohol-na{\"i}ve mice. Moreover, indicators of NET formation including citrullinated histone H3, neutrophil elastase, and neutrophil myeloperoxidase were decreased at an early time point after LPS challenge in mice receiving binge alcohol, suggesting decreased NET formation. However, in the efferocytosis phase (15 h after LPS) citrullinated histone-H3 was increased in the liver in alcohol binge mice, suggesting decreased clearance of NETs. In vitro alcohol treatment reduced efferocytosis and phagocytosis of NETotic neutrophils and promoted expression of CD206 on MΦ. Finally, depletion of neutrophils prior to binge alcohol ameliorated LPS-induced systemic inflammation and liver injury in mice. Conclusions: Dysfunctional NETosis and efferocytosis following binge drinking exacerbate liver injury associated with sepsis. Lay summary: Disease severity in alcoholic liver disease (ALD) is associated with a significant presence of neutrophils (a type of immune cell) in the liver. It remains unknown how alcohol affects the capacity of neutrophils to control infection, a major hallmark of ALD. We found that binge alcohol drinking impaired important strategies used by neutrophils to contain and resolve infection, resulting in increased liver injury during ALD.",
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T1 - Abnormal neutrophil traps and impaired efferocytosis contribute to liver injury and sepsis severity after binge alcohol use

AU - Bukong, Terence Ndonyi

AU - Cho, Yeonhee

AU - Iracheta-Vellve, Arvin

AU - Saha, Banishree

AU - Lowe, Patrick

AU - Adejumo, Adeyinka

AU - Furi, Istvan

AU - Ambade, Aditya

AU - Gyongyosi, Benedek

AU - Catalano, Donna

AU - Kodys, Karen

AU - Szabó, G.

PY - 2018/1/1

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N2 - Background & Aims: Neutrophil extracellular traps (NETs) are an important strategy utilized by neutrophils to immobilize and kill invading microorganisms. Herein, we studied NET formation and the process of neutrophil cell death (NETosis), as well as the clearance of NETs by macrophages (MΦ) (efferocytosis) in acute sepsis following binge drinking. Methods: Healthy volunteers consumed 2 ml of vodka/kg body weight, before blood endotoxin and 16 s rDNA were measured. Peripheral neutrophils were isolated and exposed to alcohol followed by phorbol 12-myristate 13-acetate (PMA) stimulation. Mice were treated with three alcohol binges and intraperitoneal lipopolysaccharide (LPS) to assess the dynamics of NET formation and efferocytosis. In vivo, anti-Ly6G antibody (IA8) was used for neutrophil depletion. Results: Inducers of NETs (endotoxin and bacterial DNA) significantly increased in the circulation after binge alcohol drinking in humans. Ex vivo, alcohol alone increased NET formation but attenuated NET formation upon PMA stimulation. Binge alcohol in mice resulted in a biphasic response to LPS. Initially, binge alcohol reduced LPS-induced NET formation and resulted in a diffuse distribution of neutrophils in the liver compared to alcohol-naïve mice. Moreover, indicators of NET formation including citrullinated histone H3, neutrophil elastase, and neutrophil myeloperoxidase were decreased at an early time point after LPS challenge in mice receiving binge alcohol, suggesting decreased NET formation. However, in the efferocytosis phase (15 h after LPS) citrullinated histone-H3 was increased in the liver in alcohol binge mice, suggesting decreased clearance of NETs. In vitro alcohol treatment reduced efferocytosis and phagocytosis of NETotic neutrophils and promoted expression of CD206 on MΦ. Finally, depletion of neutrophils prior to binge alcohol ameliorated LPS-induced systemic inflammation and liver injury in mice. Conclusions: Dysfunctional NETosis and efferocytosis following binge drinking exacerbate liver injury associated with sepsis. Lay summary: Disease severity in alcoholic liver disease (ALD) is associated with a significant presence of neutrophils (a type of immune cell) in the liver. It remains unknown how alcohol affects the capacity of neutrophils to control infection, a major hallmark of ALD. We found that binge alcohol drinking impaired important strategies used by neutrophils to contain and resolve infection, resulting in increased liver injury during ALD.

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KW - Alcoholic hepatitis

KW - Binge drinking

KW - Neutrophil depletion

KW - Neutrophil elastase

KW - Sepsis

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