A comprehensive proteomics study on platelet concentrates: Platelet proteome, storage time and Mirasol pathogen reduction technology (original) (raw)
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Transfusion Medicine and Hemotherapy, 2015
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 ...
Transfusion, 2005
BACKGROUND: Several strategies are being developed to reduce the risk of pathogen transmission associated with platelet (PLT) transfusion.STUDY DESIGN AND METHODS: The impact of a new technology for pathogen reduction based on riboflavin plus illumination (Mirasol PRT, Navigant Biotechnologies, Inc.) at 6.2 and 12.3 J per mL on functional and biochemical characteristics of PLTs was evaluated. PLT concentrates (PCs) obtained by apheresis were treated with Mirasol PRT and stored at 22°C. Modifications in major PLT glycoproteins (GPIbα, GPIV, and GPIIb-IIIa), adhesive ligands (von Willebrand factor [VWF], fibrinogen [Fg], and fibronectin), activation antigens (P-selectin and LIMP), and apoptotic markers (annexin V binding and factor [F]Va) were analyzed by flow cytometry. Adhesive and cohesive PLT functions were evaluated with well-established perfusion models. Studies were performed on the preparation day (Day 0) and during PCs storage (Days 3 and 5).RESULTS: Levels of glycoproteins remained stable during storage in PCs treated with 6.2 J per mL pathogen reduction technology (PRT) and similar to those observed in nontreated PCs. When 12.3 J per mL PRT was applied, however, levels of GPIbα moderately decreased on Days 3 and 5. VWF, Fg, and FVa were not modified in their expression levels, either by treatment or by storage period. Fibronectin appeared more elevated in all PRT samples. A progressive increase in P-selectin and LIMP expression and in annexin V binding was observed during storage of PRT-treated PCs. Functional studies indicated that 6.2 J per mL Mirasol PRT–treated PLTs preserved adhesive and cohesive functions to levels compatible with those observed in the respective control PCs.CONCLUSION: PLT function was well preserved in PCs treated with 6.2 J per mL Mirasol PRT and stored for 5 days.
Journal of Proteomics, 2010
Proteomics has brought new perspectives to the fields of hematology and transfusion medicine in the last decade. The steady improvement of proteomic technology is propelling novel discoveries of molecular mechanisms by studying protein expression, posttranslational modifications and protein interactions. This review article focuses on the application of proteomics to the identification of molecular mechanisms leading to the deterioration of blood platelets during storagea critical aspect in the provision of platelet transfusion products. Several proteomic approaches have been employed to analyse changes in the platelet protein profile during storage and the obtained data now need to be translated into platelet biochemistry in order to connect the results to platelet function.
Journal of Proteomics, 2012
Quality management of blood products is essential for blood banking. It is influenced by both processing and donor characteristics and assured by monitoring routine in vitro parameters to defined product specifications. However, these measures correlate poorly with the in vivo behavior of transfused platelets and cannot be used to select optimal donors. Since radiolabeled platelet recovery and survival studies are expensive and time consuming, there is an ongoing search for simpler measures that predict platelet transfusion outcomes. We performed a pilot study using semi-qualitative proteomics to assess changes in the platelet protein profile of donors with either acceptable or unacceptable in vivo radiolabeled autologous platelet recovery and survival measurements. Proteins changing during a 9-day storage period included cytoskeletal elements talin, vinculin and moesin as well as signal transduction proteins 14-3-3, RhoGDI and Rap1. Two of nine donations exhibited a decrease in these proteins and poor in vivo platelet recovery and survival whereas the remaining donors showed acceptable platelet recovery and survival and expected protein profiles. Analyses revealed a significant correlation between protein levels of Rap1 and RhoGDI during storage and platelet recovery and survival. This study provides for the first time preliminary data showing evidence of the utility of protein profiling to predict platelet transfusion quality. This article is part of a Special Issue entitled: Integrated omics.
Blood transfusion = Trasfusione del sangue, 2010
The Mirasol Pathogen Reduction Technology system for platelets and plasma uses riboflavin and UV light to introduce irreparable lesions into nucleic acids thereby inhibiting pathogen and white blood cell replication and reducing the load of infectious pathogens. The aim of the present study was to evaluate low plasma buffy coat platelet concentrates obtained from the OrbiSac System and to examine the effects on the development of platelet storage lesion during storage in platelet additive solution. Twenty buffy coat platelet concentrates were generated by pooling five individual units using the OrbiSac System. Riboflavin was added during the final pooling step, and the units were exposed to UV light. The bag was removed after the target energy of 6.24 J/mL had been delivered and 150 mL of platelet additive solution were added prior to storage. Platelet quality was assessed by pH, swirl, CD62P expression, lactate dehydrogenase, lactate production and glucose consumption rates over 7 ...
Transfusion and Apheresis Science, 2013
Reducing the risk of pathogen transmission to transfusion recipients is one of the great concerns in transfusion medicine. Important among the measures suggested to minimise pathogen transmission is pathogen reduction technology (PRT) systems. The present study examined the effects of Mirasol PRT system on MCS+ apheresis platelets in vitro quality measures during a seven-day storage period at 22°C. Statistical analysis indicated no significant difference in platelet concentrations between the control and treated platelet concentrates (PCs) during the storage period. Glucose and lactate levels were measured to determine metabolic activities of control and treated platelets. In both control and treated platelets, the amount of glucose consumed and lactate produced increased significantly with storage time, but glucose consumption and lactate production rates were significantly higher in treated platelets compared with control platelets. The mean pH of treated PCs was decreased at all time points relative to control PCs but remained within acceptable limits. The expression of P-selectin was also higher in Mirasol PRT treated platelets throughout the storage period, but differences were not statistically significant on Days 1 and 4. Finally, visual inspection of swirling indicated that Mirasol PRT treatment of platelets is associated with platelet shape change. Overall, our results show that MCS+ apheresis platelets treated with Mirasol PRT can preserve adequate in vitro properties for at least 5 days of storage.
Proteomics: A Tool to Study Platelet Function
International Journal of Molecular Sciences, 2021
Platelets are components of the blood that are highly reactive, and they quickly respond to multiple physiological and pathophysiological processes. In the last decade, it became clear that platelets are the key components of circulation, linking hemostasis, innate, and acquired immunity. Protein composition, localization, and activity are crucial for platelet function and regulation. The current state of mass spectrometry-based proteomics has tremendous potential to identify and quantify thousands of proteins from a minimal amount of material, unravel multiple post-translational modifications, and monitor platelet activity during drug treatments. This review focuses on the role of proteomics in understanding the molecular basics of the classical and newly emerging functions of platelets. including the recently described role of platelets in immunology and the development of COVID-19.The state-of-the-art proteomic technologies and their application in studying platelet biogenesis, s...
Vojnosanitetski pregled, 2011
Background/Aim. Bacterial contamination of blood components, primarily platelet concentrates (PCs), has been identified as one of the most frequent infectious complications in transfusion practice. PC units have a high risk for bacterial growth/multiplication due to their storage at ambient temperature (20 ? 2?C). Consequences of blood contamination could be effectively prevented or reduced by pathogen inactivation systems. The aim of this study was to determine the Mirasol pathogen reduction technology (PRT) system efficacy in PCs using an artificial bacteria-contamination model. Methods. According to the ABO blood groups, PC units (n = 216) were pooled into 54 pools (PC-Ps). PC-Ps were divided into three equal groups, with 18 units in each, designed for an artificial bacteria-contamination. Briefly, PC-Ps were contaminated by Staphylococcus epidermidis, Staphylococcus aureus or Escherichia coli in concentrations 102 to 107 colony forming units (CFU) per unit. Afterward, PC-Ps were...
Scientific Reports, 2017
Shelf life of platelet concentrates is limited to 5-7 days due to loss of platelet function during storage, commonly referred to as the platelet storage lesion (PSL). To get more insight into the development of the PSL, we used label free quantitative mass spectrometry to identify changes in the platelet proteome during storage. In total 2501 proteins were accurately quantified in 3 biological replicates on at least 1 of the 7 different time-points analyzed. Significant changes in levels of 21 proteins were observed over time. Gene ontology enrichment analysis of these proteins revealed that the majority of this set was involved in platelet degranulation, secretion and regulated exocytosis. Twelve of these proteins have been shown to reside in α-granules. Upon prolonged storage (13-16 days) elevated levels of α-2-macroglobulin, glycogenin and Ig μ chain C region were identified. Taken together this study identifies novel markers for monitoring of the PSL that may potentially also be used for the detection of "young" and "old" platelets in the circulation. Platelet transfusion is commonly used to restore platelet count and prevent bleeding in thrombocytopenic patients and patients with platelet dysfunctionality. The shelf life of platelet concentrates (PCs) is limited to a maximum storage time of 5-7 days, depending on national guidelines for platelet transfusion. Prolonged platelet storage leads to a decrease in platelet functionality known as the platelet storage lesion (PSL) 1, 2. During storage, platelet morphology changes and the discoid shape is lost 2. Furthermore, platelets become activated, as is evidenced by surface exposure of P-selectin, which is a consequence of α-granule release. PI3-kinase dependent Rap1 activation, which leads to activation of integrin αIIbβ3, has been implicated in the development of the PSL 3. In addition, levels of membrane proteins such as GPIbα and GPV decline during storage 1, 4, 5. These proteins are cleaved by ADAM17, which is dependent on p38 mitogen-activated protein kinase signaling 6. Functionality of platelets is reduced upon storage, as has been shown by a decreased response to agonists like ADP, collagen, ristocetin and PAR1 activating peptides 1, 2, 4. During storage, changes in platelet metabolism occur 7. Also an increase in levels of reactive oxygen species leading to oxidative stress has been reported 8. Despite these findings, triggers for initiating the development of the PLS are still not fully elucidated. Recent improvements in storage methods can increase platelet quality, and new platelet additive solutions seem to prolong platelet function 9, 10. However, the in vitro hallmarks of the PSL are still observed after longer storage periods 4. Several proteomic studies on stored platelets have been performed 3, 11-13. Supernatant of stored platelets 14-18 , platelet shedding 19 and platelet releasates 20 have been analyzed by mass spectrometry. Employing 2D gel/differential in gel electrophoresis (2D/DIGE), decreased levels of septin-2, β-actin and gelsolin 11 and increased levels of