Contrasting effects of stored allogeneic red blood cells and their supernatants on permeability and inflammatory responses in human pulmonary endothelial cells

Abstract

Transfusion of red blood cells (RBCs) is a common life-saving clinical practice in severely anemic or hemorrhagic patients; however, it may result in serious pathological complications such as transfusion-related acute lung injury. The factors mediating the deleterious effects of RBC transfusion remain unclear. In this study, we tested the effects of washed long-term (RBC-O; >28 days) versus short-term (RBC-F; <14 days) stored RBCs and their supernatants on lung endothelial (EC) permeability under control and inflammatory conditions. RBCs enhanced basal EC barrier function as evidenced by an increase in transendothelial electrical resistance and decrease in permeability for macromolecules. RBCs also attenuated EC hyperpermeability and suppressed secretion of EC adhesion molecule ICAM-1 and proinflammatory cytokine IL-8 in response to LPS or TNF-α. In both settings, RBC-F had slightly higher barrier protective effects as compared with RBC-O. In contrast, supernatants from both RBC-F and RBC-O disrupted the EC barrier. The early phase of EC permeability response caused by RBC supernatants was partially suppressed by antioxidant N-acetyl cysteine and inhibitor of Src kinase family PP2, while addition of heme blocker and inhibition of NOD-like receptor family pyrin domain containing protein 3 (NLRP3), stress MAP kinases, receptor for advanced glycation end-products (RAGE), or Toll-like receptor-4 (TLR4) signaling were without effect. Morphological analysis revealed that RBC supernatants increased LPS- and TNF-α-induced breakdown of intercellular junctions and formation of paracellular gaps. RBC supernatants augmented LPS- and TNF-α-induced EC inflammation reflected by increased production of IL-6, IL-8, and soluble ICAM-1. These findings demonstrate the deleterious effects of RBC supernatants on EC function, which may have a major impact in pathological consequences associated with RBC transfusion.

Transfusion‐Related acute lung injury (TRALI) caused by antibodies to HLA‐DRB1* 07:01 and HLA‐DQB1*02:02: A case report

“…4,14 Antibodymediated TRALI, the most prevalent type, is thought to be caused by the passive transfusion of antibodies from donors, which usually contain HLA I or II or human neutrophil antigen (HNA) antibodies. 13,15 Monocytes represent a primary target in the induction of TRALI by HLA-II antibodies. The HLA-II antibodies bind to cognate antigens expressed on monocytes, leading to the release of proinflammatory mediators, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, IL-8, and leukotriene B4.…”

“…11,12 The second hit is represented by two TRALI types, namely, non-antibody-mediated and antibodymediated types. 2,13 Non-antibody-mediated TRALI is thought to be caused by proinflammatory mediators, and bioactive lipids, among other factors. 4,14 Antibodymediated TRALI, the most prevalent type, is thought to be caused by the passive transfusion of antibodies from donors, which usually contain HLA I or II or human neutrophil antigen (HNA) antibodies.…”

“…The first hit occurs when the underlying clinical condition of the patient (e.g., inflammation or sepsis) activates pulmonary endothelial cells (ECs), leading to increased expression of intercellular adhesion molecule 1 (ICAM‐1) and the release of a large number of cytokines, which cause neutrophil activation, accumulation and adhesion to the activated pulmonary microvascular endothelium 11,12 . The second hit is represented by two TRALI types, namely, non‐antibody‐mediated and antibody‐mediated types 2,13 . Non‐antibody‐mediated TRALI is thought to be caused by proinflammatory mediators, and bioactive lipids, among other factors 4,14 .…”

“…Non‐antibody‐mediated TRALI is thought to be caused by proinflammatory mediators, and bioactive lipids, among other factors 4,14 . Antibody‐mediated TRALI, the most prevalent type, is thought to be caused by the passive transfusion of antibodies from donors, which usually contain HLA I or II or human neutrophil antigen (HNA) antibodies 13,15 . Monocytes represent a primary target in the induction of TRALI by HLA‐II antibodies.…”

Aberrant GPA expression and regulatory function of red blood cells in sickle cell disease

“…To measure resistance to inflammatory damage induction, endothelial cells were first incubated with RBCs for 30 minutes and then challenged with LPS (200 ng/mL). TER was then measured continuously for up to 20 hours 10 (data shown at 0, 5, 10, 15, and 20 hours).…”