Neutrophils release extracellular DNA traps during storage of red blood cell units.
Author(s): Fuchs TA, Alvarez JJ, Martinod K, Bhandari AA, Kaufman RM, Wagner DD
Publication: Transfusion, 2013, Vol. 53(12), Page 3210-6
PubMed ID: 23560771 PubMed Review Paper? No
Purpose of Paper
The purpose of this paper was to determine the effects of refrigerated storage and leukoreduction on markers of neutrophil extracellular traps (NETs).
Conclusion of Paper
The DNA content of supernatants from non-leukoreduced RBC that were refrigerated for 14-16 days or 42 days had increased by more than 100-fold and a larger DNA staining area was observed compared to plasma that was frozen without prior refrigeration. Supernatants from leukoreduced RBCs refrigerated for 42 days had significantly lower DNA, nucleosome and myeloperoxidase (MPO) content than supernatants from non-leukoreduced RBCs refrigerated for 42 days. Further, supernatants from non-leukoreduced RBCs refrigerated for 42 days contained high molecular weight DNA (>2 kb) and fragmented histone H3 which were both absent in the supernatants of leukoreduced RBCs. MPO was found to be complexed with histones. The authors report the extracellular DNA found in the supernatants of refrigerated non-leukoreduced RBCs was too large to have been derived from apoptotic cells and was not removed by filtration through a 170 uM filter as is used in transfusion.
Studies
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Study Purpose
The purpose of this study was to determine the effects of refrigerated storage and leukoreduction on markers of NETs. RBCs were collected in citrate-phosphate-dextrose (CPD) bags and stored in AS-5 or collected from the plastic tubing connected to the bags. Leukoreduction was performed on some bagged specimens within 24 h of blood collection. Bags were refrigerated for 42 days while tubing specimens were refrigerated for 14-16 days, at 4 degrees C, prior to analysis. After the refrigeration, RBC supernatants were subsequently stored at -80 degrees C prior to analysis. EDTA plasma was collected from additional individuals and stored frozen prior to analysis.
Summary of Findings:
The DNA content of supernatants from non-leukoreduced RBC that were refrigerated for 14-16 days or 42 days had increased by more than 100- fold, and a larger DNA staining area was visible in these specimens compared to plasma that was frozen without prior refrigeration. Supernatants from leukoreduced RBCs refrigerated for 42 days had significantly lower DNA, nucleosome and MPO content than supernatants from non-leukoreduced RBCs refrigerated for 42 days (p<0.001, all). Further, supernatants from non-leukoreduced RBCs refrigerated for 42 days contained high molecular weight DNA (>2 kb) and fragmented histone H3 which were absent in the supernatants of leukoreduced RBCs. MPO was found to be complexed with histones. The authors report the extracellular DNA found in the supernatants of refrigerated non-leukoreduced RBCs was too large to have been derived from apoptotic cells and was not removed by filtration through a 170 uM filter as is used in transfusion.
Biospecimens
Preservative Types
- Frozen
- Other Preservative
Diagnoses:
- Normal
Platform:
Analyte Technology Platform DNA ELISA DNA Electrophoresis DNA Light microscopy Protein ELISA Protein Light microscopy Protein Western blot Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage duration 0 days
14-16 days
42 days
Biospecimen Aliquots and Components Blood and blood products Leukoreduced red blood cells
Non-leukoreduced red blood cells
Plasma
Biospecimen Preservation Type of fixation/preservation Frozen
Refrigeration