Platelet-white blood cell (WBC) interaction, WBC apoptosis, and procoagulant activity in stored red blood cells.

Author(s): Keating FK, Butenas S, Fung MK, Schneider DJ

Publication: Transfusion, 2011, Vol. 51, Page 1086-95

PubMed ID: 21126256 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of isolation of RBCs and storage on PLT-WBC interactions, WBC apoptosis, and procoagulant activity in non-leukoreduced RBCs. The first 3 mL blood was discarded, and then 50 mL was collected into a syringe prefilled with citrate phosphate dextrose (CPD). The first sample (whole blood) was analyzed within 15 min, and the remainder of the specimen was centrifuged to obtain RBCs which were suspended in adenine, mannitol, dextrose, sodium chloride (AS-1) and analyzed immediately and after storage at 4 degrees C. Only procoagulant activity was assayed after 39 days of storage.

   Summary of Findings:

   PLT and WBC counts decreased by 10-15% and 15-20%, respectively between day 2 and 15 of storage at 4 degrees C. Monocytes became harder to discriminate during storage and neutrophils exhibited increased leftward shift in side scatter. PLA and PLT-monocyte aggregates (PMA) increased from 1.6% and 7.2%, respectively, in whole blood to 3.4% and 47.1%, respectively, in fresh RBCs, but only the increase in PMA was significant (p<0.01). PLAs remained high during storage, and when normalized to neutrophils, PLAs were higher on days 3-15 than on day 1, but when normalized to monocytes, they were at similar levels after 1 day and 15 days of storage. Neutrophil and monocyte apoptosis, measured by annexin V binding, were <1% and <5% in whole blood and fresh RBCs, respectively, but after 15 days of storage, annexin V binding was observed for 94.8% of neutrophils and 72.6% of monocytes. Neutrophil and monocyte cell death, as measured by 7-AAD binding, was <1% in fresh RBCs and <4% after 5 days, but cell death levels increased to 24.3% and 33.96% of cells, respectively after 15 days of storage. Because annexin V binding was observed earlier and in a higher percentage of cells, the authors conclude WBC cell death during storage is predominantly due to apoptosis. While <1% of PLA in fresh RBCs contained WBC costained with 7-AAD, after 15 days of storage, 57% of neutrophils and 28% of monocytes in aggregates costained with 7-AAD, and annexin V was localized to some PLA after 5 days storage. TF mean fluorescence increased from 24 in fresh RBCs to 54 in RBCs stored 15 days (p=0.002). The supernatant of fresh RBCs had no procoagulant activity, but procoagulant activity increased with storage and was detected after 5-12 days of storage (depending on donor), but could be attenuated by centrifugation of the supernatant before assay or addition of a TF inhibitory antibody.

   Biospecimens

   • Bodily Fluid - Blood
   • Cell - Red Blood Cells

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Cell count/volume	Flow cytometry
   Cell count/volume	Hematology/ auto analyzer
   Cell count/volume	Fluorescent microscopy
   Morphology	Hematology/ auto analyzer

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 days 1 day 3 days 5 days 10 days 15 days 39 days
   Biospecimen Aliquots and Components	Blood and blood products	Red blood cells Whole blood

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