Abstract
Red blood cells (RBCs) have been shown to affect immune function and can induce inflammatory responses after transfusion. The transfusion of washed RBCs can significantly reduce adverse effects, however, the soluble factors that may mediate these effects have not been identified. Previous studies have identified, but not quantified, a small number of chemokines associated with RBCs. We isolated RBCs from healthy volunteers and quantified of a panel of 48 cytokines, chemokines, and growth factors in the lysate, cytosol, and conditioned media of these cells using Luminex® technology. This analysis revealed that, after correcting for white blood cell and platelet contamination, 46 cytokines were detected in RBC lysates, and the median concentration in RBCs was 12-fold higher than in the plasma. In addition, extensive washing of RBCs, such as that performed in proteomics analyses or prior to some RBC transfusions, significantly attenuated the release of six cytokines following incubation at 37 °C. This supports the hypothesis that, alongside its gas exchange function, RBCs play a role in cytokine signalling. This discovery may help supplement disease biomarker research and may shed light on adverse inflammatory processes that can follow RBC transfusion.
Highlights
Red blood cells (RBCs) have been shown to affect immune function and can induce inflammatory responses after transfusion
In a recent study from our laboratory, it was determined that the pro-inflammatory cytokine and enzyme, macrophage migration inhibitory factor (MIF), was present in RBCs at levels 1000-fold higher than the typical plasma concentration[7]
To identify if there were more cytokines associated with RBCs and to quantify the levels, the cytokine profile of RBCs, white blood cells, platelets, and plasma was determined using an antibody-mediated multiplex bead array
Summary
Red blood cells (RBCs) have been shown to affect immune function and can induce inflammatory responses after transfusion. Extensive washing of RBCs, such as that performed in proteomics analyses or prior to some RBC transfusions, significantly attenuated the release of six cytokines following incubation at 37 °C This supports the hypothesis that, alongside its gas exchange function, RBCs play a role in cytokine signalling. Differential white blood cell counts and morphological changes in leukocytes can provide valuable diagnostic information that supplement other tests and clinical assessments[3] These cells are known to produce and respond to a number of cytokines to either promote or suppress inflammatory processes[4,5]. RBCs, or soluble factors released by these cells, can stimulate the secretion of pro-inflammatory markers from lung fibroblasts[8] These studies demonstrate that RBCs may be an important component of the immune system and are capable of signalling or receiving signal from other cell types. This analysis included identifying the total concentration in RBC lysates and the concentration released by intact RBCs and investigation into how the cytokine profile can be modulated
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