Transition to 37°C reveals importance of NADPH in mitigating oxidative stress in stored RBCs
Aline Roch, Nicholas J. Magon, Jessica Maire, Cacang Suarna, Anita Ayer, Sophie Waldvogel, Beat A. Imhof, Mark J. Koury, Roland Stocker, Marc Schapira
Aline Roch, Nicholas J. Magon, Jessica Maire, Cacang Suarna, Anita Ayer, Sophie Waldvogel, Beat A. Imhof, Mark J. Koury, Roland Stocker, Marc Schapira
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Research Article Hematology Metabolism

Transition to 37°C reveals importance of NADPH in mitigating oxidative stress in stored RBCs

Abstract

The RBC storage lesion is a multiparametric response that occurs during storage at 4°C, but its impact on transfused patients remains unclear. In studies of the RBC storage lesion, the temperature transition from cold storage to normal body temperature that occurs during transfusion has received limited attention. We hypothesized that multiple deleterious events might occur in this period of increasing temperature. We show dramatic alterations in several properties of therapeutic blood units stored at 4°C after warming them to normal body temperature (37°C), as well as febrile temperature (40°C). In particular, the intracellular content and redox state of NADP(H) were directly affected by post-storage incubation at 37°C, as well as by pro-oxidant storage conditions. Modulation of the NADPH-producing pentose phosphate pathway, but not the prevention of hemoglobin autoxidation by conversion of oxyhemoglobin to carboxyhemoglobin, provided protection against storage-induced alterations in RBCs, demonstrating the central role of NADPH in mitigating increased susceptibility of stored RBCs to oxidative stress. We propose that assessing RBC oxidative status after restoration of body temperature constitutes a sensitive method for detecting storage-related alterations that has the potential to improve the quality of stored RBCs for transfusion.

Authors

Aline Roch, Nicholas J. Magon, Jessica Maire, Cacang Suarna, Anita Ayer, Sophie Waldvogel, Beat A. Imhof, Mark J. Koury, Roland Stocker, Marc Schapira

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Figure 4

Effects of oxygen removal by CO purging on oxidative stress and osmotic fragility after warming to 37°C of RBCs previously stored for 2 weeks.

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Effects of oxygen removal by CO purging on oxidative stress and osmotic ...
(A) Hemoglobin absorption spectra (OD) of untreated (solid line) or CO-treated RBCs analyzed 1 hour (dashed) or 2 weeks (dotted) after treatment. (B) Formation of H2O2 by RBCs without (diamonds) or with CO treatment (triangles). RBCs were stored for 0 or 2 weeks at 4°C, followed by 20 hours at 37°C at 5% hematocrit (n = 3). (C) Percent Prx2 dimerization (n = 6) and (D) osmotic fragility based on solution osmolality leading to 50% hemolysis (n = 6) in untreated (white) or CO-treated (gray) blood samples. RBCs were stored for 0 or 2 weeks at 4°C, followed by 20 hours at 37°C. Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). Bars show mean values. *P ≤ 0.05, repeated-measures 2-way ANOVA; #P ≤ 0.05, Mann-Whitney nonparametric test.