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Isoprostane and isofuran lipid mediators accumulate in stored red blood cells and influence platelet function in vitro.

Author(s): Spinelli SL, Lannan KL, Casey AE, Croasdell A, Curran TM, Henrichs KF, Pollock SJ, Milne GA, Refaai MA, Francis CW, Phipps RP, Blumberg N

Publication: Transfusion, 2014, Vol. 54, Page 1569-79

PubMed ID: 24192515 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 RBC leukoreduction and storage duration on the accumulation of F2-IsoPs and IsoFs and on plt function. Additive solution 1 (AS-1) RBCs were obtained from a blood bank and leukoreduced with a leukoreduction filter. RBC supernatants were obtained by centrifugation. Some RBCs were washed after 13 or 26 days of storage in an isotonic solution and then stored for another 24 h at 4 degrees C. Plts were obtained by centrifugation of sodium citrate blood from healthy blood donors and were resuspended in Tyrode's/acid citrate dextrose (ACD) with 0.1 ug/mL prostacyclin. Plt supernatants were stored frozen at -80 degrees C.

   Summary of Findings:

   During refrigerated storage of non-leukoreduced RBCs, sCD40L levels increased. During storage of leukoreduced RBCs, levels of F2-IsoPs, IsoFs and hemoglobin increased linearly (R2=0.9632, R2=0.8816 and R2=0.9936, respectively). Non-leukoreduced RBCs were not evaluated. Further, the ability of leukoreduced RBC supernatants to induce TXB2 and sCD40L release from Plts increased after 19 and 40 days of refrigeration, respectively (p<0.05, both), and CD62P expression on Plts increased after 40 days of refrigerated storage (p<0.05). While increasing the concentration of 8-Iso-PGF led to increased Plt release of TXB2 and expression of CD62P, it did not affect sCD40L release. Leukoreduction increased levels of TXB2 in RBCs stored <8 or >20 days (p<0.05, both), decreased levels of sCD40L in RBCs stored <8 or >20 days (p<0.05, both), and increased PGE2 in RBCs stored for >20 days only (p<0.05). Additionally, the authors report a reduction in vascular endothelial growth factor (VEGF) after leukoreduction of RBCs. Supernatants of non-leukoreduced RBCs refrigerated for <8 or >20 days induced more Plt aggregation than supernatants of fresh RBCs, but the supernatants of leukoreduced refrigerated RBCs induced comparable aggregation to fresh RBCs. Washing RBCs after storage reduced F2-IsoPs and IsoFs (p<0.05 for those stored 13 or 26 days), and while levels increased during the 24 h post wash storage, they remained significantly lower than pre-wash levels (p<0.05 for those stored 13 or 26 days).

   Biospecimens

   • Cell - Red Blood Cells

   Preservative Types

   • Other Preservative
   • Frozen

   Diagnoses:

   • Normal

   Platform:

   Analyte	Technology Platform
   Small molecule	GC-MS
   Cell count/volume	Flow cytometry
   Lipid	Enzyme assay
   Protein	ELISA
   Protein	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	0 h 24 h 5 days <8 days 12 days 19 days >20 days 26 days 33 days 40 days 47 days
   Biospecimen Aliquots and Components	Blood processing method	Leukoreduced Non-leukoreduced Washed Not washed
   Analyte Extraction and Purification	Washing	Washed Unwashed
   Biospecimen Preservation	Type of fixation/preservation	Refrigeration Frozen

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