Neutrophil extracellular traps in stored canine red blood cell units

Abstract Background Neutrophil extracellular traps (NETs), webs of DNA and citrullinated histones extruded from activated neutrophils cause transfusion‐related acute lung injury. Supernatants of stored red blood cell (RBC) units might promote NETosis in neutrophils from the units or from transfusion...

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Main Authors: Erin R. McQuinn, Stephanie A. Smith, Austin K. Viall, Chong Wang, Dana N. LeVine
Format: Article
Language:English
Published: Wiley 2020-09-01
Series:Journal of Veterinary Internal Medicine
Subjects:
Online Access:https://doi.org/10.1111/jvim.15876
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author Erin R. McQuinn
Stephanie A. Smith
Austin K. Viall
Chong Wang
Dana N. LeVine
author_facet Erin R. McQuinn
Stephanie A. Smith
Austin K. Viall
Chong Wang
Dana N. LeVine
author_sort Erin R. McQuinn
collection DOAJ
description Abstract Background Neutrophil extracellular traps (NETs), webs of DNA and citrullinated histones extruded from activated neutrophils cause transfusion‐related acute lung injury. Supernatants of stored red blood cell (RBC) units might promote NETosis in neutrophils from the units or from transfusion recipients. Hypotheses (1) NETs form during storage of canine RBC, (2) leukoreduction (LR) before storage of RBC reduces NETosis, and (3) supernatant from stored, nonleukoreduced (NLR) RBC units induces NETosis in healthy canine neutrophils modeling transfusion recipients. Animals Six healthy purpose‐bred research dogs were utilized for blood donation. Methods Prospective controlled study. RBC units were collected from each dog, aseptically divided into 2 equal subunits, 1 of which was leukoreduced, and stored for 42 days. Stored units were sampled biweekly for quantification of NET markers citrullinated histone H3 (Western blot) and cell‐free DNA (cfDNA) (DNA dye binding). Unit supernatants were applied ex vivo to canine neutrophils and extracellular DNA release representing NETosis was assessed. Results Markers of NETs increased during RBC storage (cfDNA P < .0001 and citrullinated H3 P = .0002) and were higher in NLR than LR units (day 42 LR cfDNA 0.34 ± 0.82 ng/mL vs day 42 NLR 1361.07 ± 741.00 ng/mL, P < .0001; day 42 LR citrullinated H3 0.19 ± 0.13 AU vs NLR 0.57 ± 0.34 AU, P = .007). Isolated neutrophils did not form NETs when exposed to stored canine RBC supernatant. Conclusions and Clinical Importance NETosis occurs in stored canine NLR RBC units, and is attenuated by LR before storage. NETs might be mediators of transfusion reactions.
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spelling doaj.art-b5b273388dc24d1fb6f18ad6bdf67f862022-12-22T03:00:57ZengWileyJournal of Veterinary Internal Medicine0891-66401939-16762020-09-013451894190210.1111/jvim.15876Neutrophil extracellular traps in stored canine red blood cell unitsErin R. McQuinn0Stephanie A. Smith1Austin K. Viall2Chong Wang3Dana N. LeVine4Department of Veterinary Clinical Sciences Iowa State University Ames Iowa USADepartment of Biochemistry, College of Medicine University of Illinois at Urbana‐Champaign Urbana Illinois USADepartment of Veterinary Pathology Iowa State University Ames Iowa USADepartment of Veterinary Diagnostic and Production Animal Medicine Iowa State University Ames Iowa USADepartment of Veterinary Clinical Sciences Iowa State University Ames Iowa USAAbstract Background Neutrophil extracellular traps (NETs), webs of DNA and citrullinated histones extruded from activated neutrophils cause transfusion‐related acute lung injury. Supernatants of stored red blood cell (RBC) units might promote NETosis in neutrophils from the units or from transfusion recipients. Hypotheses (1) NETs form during storage of canine RBC, (2) leukoreduction (LR) before storage of RBC reduces NETosis, and (3) supernatant from stored, nonleukoreduced (NLR) RBC units induces NETosis in healthy canine neutrophils modeling transfusion recipients. Animals Six healthy purpose‐bred research dogs were utilized for blood donation. Methods Prospective controlled study. RBC units were collected from each dog, aseptically divided into 2 equal subunits, 1 of which was leukoreduced, and stored for 42 days. Stored units were sampled biweekly for quantification of NET markers citrullinated histone H3 (Western blot) and cell‐free DNA (cfDNA) (DNA dye binding). Unit supernatants were applied ex vivo to canine neutrophils and extracellular DNA release representing NETosis was assessed. Results Markers of NETs increased during RBC storage (cfDNA P < .0001 and citrullinated H3 P = .0002) and were higher in NLR than LR units (day 42 LR cfDNA 0.34 ± 0.82 ng/mL vs day 42 NLR 1361.07 ± 741.00 ng/mL, P < .0001; day 42 LR citrullinated H3 0.19 ± 0.13 AU vs NLR 0.57 ± 0.34 AU, P = .007). Isolated neutrophils did not form NETs when exposed to stored canine RBC supernatant. Conclusions and Clinical Importance NETosis occurs in stored canine NLR RBC units, and is attenuated by LR before storage. NETs might be mediators of transfusion reactions.https://doi.org/10.1111/jvim.15876cfDNAhistoneleukoreductiontransfusion reaction
spellingShingle Erin R. McQuinn
Stephanie A. Smith
Austin K. Viall
Chong Wang
Dana N. LeVine
Neutrophil extracellular traps in stored canine red blood cell units
Journal of Veterinary Internal Medicine
cfDNA
histone
leukoreduction
transfusion reaction
title Neutrophil extracellular traps in stored canine red blood cell units
title_full Neutrophil extracellular traps in stored canine red blood cell units
title_fullStr Neutrophil extracellular traps in stored canine red blood cell units
title_full_unstemmed Neutrophil extracellular traps in stored canine red blood cell units
title_short Neutrophil extracellular traps in stored canine red blood cell units
title_sort neutrophil extracellular traps in stored canine red blood cell units
topic cfDNA
histone
leukoreduction
transfusion reaction
url https://doi.org/10.1111/jvim.15876
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AT chongwang neutrophilextracellulartrapsinstoredcanineredbloodcellunits
AT dananlevine neutrophilextracellulartrapsinstoredcanineredbloodcellunits