Microparticles in red cell concentrates prime polymorphonuclear neutrophils and cause acute lung injury in a two-event mouse model

doi:10.1016/j.intimp.2017.11.029

International Immunopharmacology

Volume 55, February 2018, Pages 98-104

https://doi.org/10.1016/j.intimp.2017.11.029 Get rights and content

Highlights

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Microparticles accumulated in stored RCCs
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Isolated MPs and the RCC supernatant primed the PMN respiratory burst.
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Microparticles in RCCs induced mouse ALI.

Abstract

Red cell-derived microparticles (RMPs) are potential mediators of transfusion-related acute lung injury (TRALI). The aim of this study was to investigate the effects of microparticles present in red cell concentrates (RCC) on polymorphonuclear neutrophil (PMN) respiratory burst and acute lung injury (ALI) in mice. Microparticles (MPs) in RCC supernatant were quantified using flow cytometry. The priming activity of either isolated MPs or RCC supernatant toward human PMN was measured in vitro. Mice were injected with lipopolysaccharide (LPS), followed by an infusion of either isolated MPs or heat-treated RCC supernatant. The lungs were harvested to assess myeloperoxidase (MPO) activity, histology and pulmonary edema. Protein content in bronchoalveolar lavage fluid (BALF) was measured. The number of RMPs increased significantly during storage. Both isolated MPs and the supernatants from RCCs that had been stored for 28 and 35 days effectively primed the PMN respiratory burst. The infusion of isolated MPs or supernatants that had been stored for > 28 days into LPS-treated mice caused ALI. The filtered supernatant resulted in significantly ameliorated mouse ALI. MPs that accumulate during RCC storage prime the PMN respiratory burst and cause ALI in a two-event mouse model.

Introduction

Transfusion-related acute lung injury (TRALI) is a major cause of transfusion-related mortality. Causative factors are categorized into antibody- and non-antibody-mediated TRALI. The transfusion of stored red cell concentrates (RCCs) may be related to various adverse reactions, including TRALI [1], [2], [3]. During RCC storage, many types of storage-related events may occur, including decreases in adenosine 5′-triphosphate (ATP) levels and the depletion of 2,3-diphosphoglycerate (2,3-DPG), both of which accompany the loss of membrane integrity and the release of microvesicles into the additive solution. These microvesicles, also known as microparticles, are surrounded by a lipid bilayer and contain phospholipids, membrane-associated proteins, enzymes and chemokines; furthermore, these particles have been identified to exhibit proinflammatory and procoagulant activities [4], [5], [6]. Red blood cell-derived microparticles (RMPs) are enriched in hemoglobin, complement C complex, band 3 neo-antigens and immunoglobulin G, all of which are eliminated from aging red blood cells [7], [8]. These components may contribute to multiple effects, such as thrombin generation [9], [10], nitric oxide (NO) depletion [11], immunosuppression [12], [13] and inflammation [5], among others.

Maslanka et al. [14] investigated 464 blood components (RBCs, PLTs, L-PLTs, and FFP) in 271 patients who experienced adverse post-transfusion reactions and revealed that microparticles released from stored red blood cells are a potential factor contributing to non-antibody-mediated TRALI. According to the two-event TRALI model, excessive neutrophil activation plays a central role in the pathogenesis of TRALI [15]. Early studies suggested that platelet microparticles carried P-selectin, which bound and activated neutrophils [16], and that enriched complement C in RMPs could activate neutrophils via the Fc receptors expressed on the surface [17]. Cardo et al. [18] demonstrated that the ability of PRBC supernatants to prime neutrophils was also present in microvesicles. Our previous study also indicated that platelet microparticles could prime the neutrophil respiratory burst and exacerbate damage to human pulmonary microvascular endothelial cells [19]. Although the ability of microparticles to prime neutrophils has already been described, the role and mechanism of MPs in the development of TRALI have not been elucidated.

In this study, we investigated the priming activity of MPs from stored RCCs and their capability to cause acute lung injury (ALI) in mice in a lipopolysaccharide (LPS)-treated two-event model.

Section snippets

Sample preparation

Whole blood was collected using a polyvinyl chloride collection bag system (Shanghai Transfusion Technology Co. Ltd., Shanghai, China) with an anticoagulant citrate dextrose solution B (ACD-B, Shanghai Transfusion Technology Co. Ltd., Shanghai, China). After the blood was stored overnight, the plasma was separated via centrifugation at 3820 × g for 20 min, after which a mannitol-adenine-phosphate additive solution (MAP, Shanghai Transfusion Technology Co. Ltd., Shanghai, China) was added to a

RMPs accumulated during RCC storage

Five units of RCCs were stored under standard blood bank conditions, and samples were withdrawn on days 1, 7, 14, 21, 28, and 35. RMPs were identified as a closed vesicle positive for CFSE fluorescence and the red cell membrane-specific protein CD235a as assessed using flow cytometry (Fig. 1). The number of RMPs was significantly increased after 28 days of storage; this number continued to climb until it reached an 18-fold increase on day 35 compared to the number on day one. The levels of RMPs

Discussion

Studies have demonstrated that red blood cell-derived microparticles (RMPs) may accumulate during the storage of red blood cell products, and the amount accumulated is dependent on the blood component processing method, additive solution and storage time. Although pre-storage leukocyte depletion has been reported to decrease the formation of MPs in stored RCCs and PLTs [18], [24], universal leukoreduction processing has still not been achieved in many developing and underdeveloped countries. In

Research funding

This work was supported by grants from the Shanghai Natural Science Foundation (Grant no. 15ZR1438300), the Shanghai Public Health Construction Projects Foundation (Grant No. 15GWZK0501) and the Foundation of Shanghai Municipal Commission of Health and Family Planning (Grant nos. 201440439, 201640096, 20154Y0192 and 20154Y0079).

Acknowledgments

We would like to thank the Department of Animal Science at the Shanghai Jiaotong University School of Medicine for performing laboratory animal husbandry.

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^☆: The authors certify that they have no affiliation with or financial involvement in any organization or entity with a direct financial interest in the subject matter or materials discussed in this manuscript.

¹: These authors contributed equally to this article.

View full text

Microparticles in red cell concentrates prime polymorphonuclear neutrophils and cause acute lung injury in a two-event mouse model☆

Highlights

Abstract

Introduction

Section snippets

Sample preparation

RMPs accumulated during RCC storage

Discussion

Research funding

Acknowledgments

Transfus. Apher. Sci.

Blood Cells Mol. Dis.

Redox Biol.

Blood

Transfus. Apher. Sci.

Blood

J. Am. Coll. Surg.

Pathology

Age of blood and recipient factors determine the severity of transfusion-related acute lung injury (TRALI)

Crit. Care

Dissimilar respiratory and hemodynamic responses in TRALI induced by stored red cells and whole blood platelets

Blood

Bad blood: the risks of red cell storage

Nat. Med.

Platelets amplify inflammation in arthritis via collagen-dependent microparticle production

Science

Red blood cell microparticles: clinical relevance

Transfus. Med. Hemother.

Erythrocyte vesiculation: a self-protective mechanism?

Br. J. Haematol.

The proteome of red cell membranes and vesicles during storage in blood bank conditions

Transfusion