Packed blood cells stored in AS-5 become proinflammatory during storage.
BACKGROUND
Studies have shown that packed blood cells (PBCs) stored in AS-1 (Adsol, Baxter) and AS-3 (Nutricel, Medsep Corp.) accumulate proinflammatory substances, which may contribute to increased complications from allogeneic blood transfusion. This study assessed whether supernates from PBCs stored in AS-5 (Optisol, Terumo Corp.) prime neutrophils (PMNs), activate platelets (PLTs), and accumulate proinflammatory cytokines and PMN granule constituents.


METHODS
PBC units were prepared in AS-5 from nonleukoreduced (NLR) and leukoreduced (LR) whole-blood units and stored at 4 degrees C. Supernates from samples of PBCs collected at various storage times were analyzed by multiplex enzyme-linked immunosorbent assay for proinflammatory cytokines and myeloperoxidase (MPO) and were incubated with type-matched blood, which was assessed by flow cytometry for expression of CD11b on PMNs, CD62P on PLTs, and formation of PMN-PLT aggregates.


RESULTS
Supernates from NLR PBCs stored for at least 14 days elevated CD11b expression on PMNs and the number of PMN-PLT aggregates compared to supernates from collection day PBCs. The magnitude of these effects correlated with storage age. Supernates from LR PBCs did not elicit these responses. Expression of CD62P on PLTs was not affected by supernates from either NLR or LR PBCs. Levels of interleukin (IL)-1beta, IL-6, IL-8, IL-18, NAP-2, MCP-1, RANTES, and MPO were elevated in supernates from 28- and 42-day NLR units. Tumor necrosis factor alpha and MIP-1alpha did not increase, and cytokine levels in LR PBC units did not increase.


CONCLUSIONS
Units of NLR PBCs stored in AS-5 become increasingly proinflammatory as a function of storage time. Leukoreduction prevents this change.
Department of Blood Research, Division of Military Casualty Research, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD 20910, USA. steve.mcfaul@us.army.mil
Steve J
SJ
McFaul
Jason B
JB
Corley
Craig W
CW
Mester
Jayasree
J
Nath
D001773:Blood Cells / Q000276:immunology* / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism; D001793:Blood Preservation; D039481:CD11b Antigen / Q000378:metabolism; D018932:Chemokine CCL2 / Q000378:metabolism; D018946:Chemokine CCL5 / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D020382:Interleukin-18 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism
C111835:CCL2 protein, human; D039481:CD11b Antigen; D018932:Chemokine CCL2; D018946:Chemokine CCL5; C509295:ITGAM protein, human; D020382:Interleukin-18; D015850:Interleukin-6; D016209:Interleukin-8; C104356:NAP1L4 protein, human; D009687:Nuclear Proteins; D019007:P-Selectin