Abstract
Context: Neutrophils are the primary effector cells in the pathogenesis of transfusion-related acute lung injury or multiple organ failure after blood transfusion.Objective: We aimed to investigate the effect of fresh (1 day preparation) and aged (42 day preparation) PRBC-derived plasma on neutrophil morphology, migration and phagocytosis.Materials and methods: We evaluated the production of reactive oxygen species (ROS) and the expression of non-muscle myosin heavy chain IIA (MYH9) in neutrophils treated with PRBC-derived plasma. We used western blots and antibody arrays to evaluate changes in signal transduction pathways in plasma-treated neutrophils.Results: Aged PRBC-derived plasma elicited a stronger oxidative burst in neutrophils when compared with fresh PRBC-derived plasma (p < 0.05). Antibody arrays showed increased phosphorylation of NF-ĸB proteins (p105, p50 and Ikk) in aged PRBC-derived plasma-treated neutrophils. The expression of non-muscle myosin IIA (MYH9), a cytoskeleton protein involved in immune cell migration and morphological change, was also significantly upregulated in neutrophils treated with aged PRBC-derived plasma compared to fresh plasma (p < 0.05). Pretreatment of neutrophils with blebbistatin (a specific type II myosin inhibitor), ascorbic acid (an antioxidant), or staurosporine (a protein tyrosine kinase inhibitor), effectively abrogated the morphological changes, neutrophil migration, and phagocytosis induced by aged PRBC-derived plasma.Conclusion: Upregulation of MYH9 in neutrophils treated with aged PRBC-derived plasma and abrogation of neutrophil migration in blebbistatin-treated neutrophils suggested a functional role of MYH9 in the directional migration of immune cells. Our data help elucidate the cellular and molecular mechanisms of transfusion-related injury.
Highlights
Neutrophils are the main effector cells of inflammation in the airway epithelium.[1]
There was an increase in superoxide production when neutrophils were incubated with the different packed red blood cell (PRBC)-derived plasma preparations, suggesting that PRBC-derived plasma induced an oxidative burst in human neutrophils. fMLP was used as a positive control and untreated neutrophils were used as a control
Since oxidative burst triggered by UV or cytokines is known to induce protein tyrosine phosphorylation, we evaluated the effect of plasma on protein tyrosine phosphorylation in neutrophils
Summary
Neutrophils are the main effector cells of inflammation in the airway epithelium.[1] Neutrophil priming has been reported to play a crucial role in the pathogenesis of post-injury hyper-inflammation leading to transfusionrelated acute lung injury (TRALI) and multiple organ failure (MOF) in surgery patients.[2] the pathogenesis of TRALI has been linked to the presence of antileukocyte antibodies, a significant number of cases do not have an immunologic etiology,(3) leading to the postulation of the two-event model for TRALI. The two-event model suggests that TRALI is initiated by an immune priming step (consisting of the initial trauma), followed by exposure of neutrophils to specific biological response modifiers via transfusion, causing an interaction between neutrophils, platelets and lung Abbreviations. STAT the Signal Transducers and Activators of Transcription protein.
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call