Treatment of Platelet Concentrates with the Mirasol Pathogen Inactivation System Modulates Platelet Oxidative Stress and NF-κB Activation

Abstract

Background: Pathogen inactivation (PI) technologies for platelets aim to improve transfusion safety by preventing the replication of contaminating pathogens. However, as a consequence of treatment, aspects of the platelet storage lesion are amplified. Mirasol treatment also affects platelet signal transduction and apoptotic protein expression. The aim of this study was to examine the effect of Mirasol treatment on the generation of reactive oxygen species (ROS) and subsequent oxidative stress. Methods: Pooled platelet concentrates were prepared in platelet-additive solution (70% SSP+ / 30% plasma). ABO-matched platelets were pooled and split, and treated with the Mirasol system (TerumoBCT) or left untreated as a control. Platelet samples were tested on day 1, 5, and 7 post-collection. Results: Mirasol-treated platelets had increased formation of ROS by day 5 of storage. Oxidative damage, in the form of protein carbonylation, was higher in Mirasol-treated platelets, whilst no effect on nitrotyrosine formation or lipid peroxidation was detected. The NF-κB signaling pathway was also activated in Mirasol-treated platelets, with increased expression and phosphorylation of NF-κB p65 and IκBa. Conclusion: These data demonstrate that Mirasol-treated platelets produce more ROS and display protein alterations consistent with oxidative damage.

p38 mitogen‐activated protein kinase regulates mitochondrial function and microvesicle release in riboflavin‐ and ultraviolet light–treated apheresis platelet concentrates

“…19,33 Previously, several studies have reported that RF and UV light can trigger intracellular generation of ROS in PLTs modulating signaling pathways and causing oxidative damage of macromolecules. 20,21,34 In this study, we also show that the RF/UV treatment generated intracellular H 2 O 2 in apheresis PCs throughout storage, which might contribute to the depolarization of the DW m further contributing to apoptosis development.…”

“…Accumulating evidence demonstrates that PI-treated PLTs share highly similar behaviors with PLTs stimulated by physiologic activators, including PLT activation, degranulation, and development of apoptosis, leading to the conclusion that these processes have the same signaling mechanism. Lopez and coworkers 30 reported that thrombin induces translocation of Bax, Bak, and active Bid to MT resulting in cyto c release and subsequent loss 20,21,34 In this study, we also show that the RF/UV treatment generated intracellular H 2 O 2 in apheresis PCs throughout storage, which might contribute to the depolarization of the DW m further contributing to apoptosis development. …”

“…19 In addition, RF/UV treatment of blood components triggers ROS generation resulting in oxidative damage of plasma proteins and mitochondrial DNA (mtDNA). 20,21 Although antioxidant systems present in cells and PLTs can function as scavengers to inactivate the excess intracellular ROS, high levels of ROS can result in the destruction of macromolecules and lead to cell death. The p38 mitogen-activated protein kinase (p38 or p38 MAPK) is a stress kinase belonging to the mitogenactivated kinase superfamily; it regulates several signaling cascades involved in cell division, differentiation, and apoptosis.…”

Redox Proteomics and Platelet Activation: Understanding the Redox Proteome to Improve Platelet Quality for Transfusion

“…Supporting this notion, Johnson et al found higher levels of carbonylation in Mirasol ® -treated PCs compared to untreated buffy coat-derived PCs [20]. Carbonyl insertion into proteins occurs mainly by three mechanisms: amino acid backbone scission, amino acid side chain cleavage, and side chain ketone or aldehyde formation by metal catalysis [46].…”

Pathogen-Inaktivierungssysteme für Thrombozytenkonzentrate

“…Der Riboflavinabbau ist zeitabhängig [98]. Umfangreiche toxikologische Untersuchungen sind unternommen worden, um die Sicherheit der mit MIRASOL® behandelten Blutprodukte nachzuweisen (Übersicht in [69,70] [17,115,116]. Dennoch finden sich nach einer PI mit MIRASOL® nur geringe Veränderungen am Gesamtproteom [67].…”