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 1

Effects of incubation at 37°C on physical properties, Prx2 dimerization, and NADP(H) content of RBCs previously stored at 4°C.

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Effects of incubation at 37°C on physical properties, Prx2 dimerization,...
(A) Osmotic fragility based on solution osmolality leading to 50% hemolysis (n = 6); and (B) elongation index (measure of RBC deformability) (n = 6) at 0, 2, 4, and 6 weeks of storage at 4°C before (white) and after warming to 37°C for 4 hours (light gray) or 20 hours (dark gray). (C) Hydrogen peroxide formation as described in Methods during 20 hours at 4°C or 37°C (5% hematocrit) after storage for 0, 2, 4, or 6 weeks at 4°C (n = 6). (D) Percentage Prx2 dimerization (n = 6) at 0, 2, 4, and 6 weeks of storage at 4°C before (white) and after warming to 37°C for 4 hours or 20 hours. (E) Total NADP(H) and (F) NADPH determined by enzymatic cycling at 0, 2, 4, and 6 weeks of storage at 4°C before and after warming to 37°C for 24 hours (n = 6). Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). *P ≤ 0.05, repeated-measures 2-way ANOVA. See Supplemental Figure 5 and full uncut gels.