Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Towards a detailed understanding of the red blood cell storage lesion: and its consequences for in vivo survival following transfusion
Umeå University, Faculty of Medicine, Department of Community Medicine and Rehabilitation, Idrottsmedicin. Umeå University, Faculty of Medicine, Department of Surgical and Perioperative Sciences. Umeå University, Faculty of Medicine, Department of Integrative Medical Biology (IMB).
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Red blood cells (RBCs) are vital for oxygen delivery to tissues and constitute the vast majority of all cells in blood. After leaving the red bone marrow as mature cells, RBCs have a lifespan of approximately 120 days before they are removed from the circulation by macrophages, mainly in the spleen and liver. RBC transfusion is a common therapy in modern healthcare. Major surgery, numerous cancer treatments and other, often lifesaving, interventions would be unthinkable without available blood supply. For this reason, hospitals store donated RBCs in blood banks.

The metabolic and structural changes that occur during prolonged storage of RBCs (the storage lesion) have been studied in detail in vitro and include oxidative stress, a reduction in glycolysis, increased membrane rigidity and shedding of microparticles from the RBC membrane. Stored RBCs share several features of senescent RBCs, but also with RBCs undergoing an apoptotic-like process called eryptosis. A consequence of the storage lesion is the fact that as much as 25% of stored RBCs could be rapidly removed from the circulation within 24 hours after transfusion. The mechanisms behind this rapid macrophage-mediated recognition and removal of stored RBCs, and its immunological consequences, remain largely unknown. Therefore, the aims of this thesis were to investigate if cryopreserved human RBCs induced an inflammatory response following autologous transfusion into healthy volunteers, and to further understand the mechanisms behind macrophage recognition of stored RBCs in vitro and in vivo.

Autologous transfusion of two units of cryopreserved RBCs into healthy human recipients was found to be associated with an increased extravascular RBC elimination already at 2 hours after transfusion. However, there were no signs of an increased production of any of the investigated pro-inflammatory cytokines, indicating that an increase in the destruction of RBCs per se did not induce an inflammatory response.

Eryptosis is a form of induced RBC death associated with an increased cytoplasmic Ca2+ uptake. We found that a subset of human RBCs increased their Ca2+ permeability during prolonged storage at +4°C. Using a murine model, to further understand how RBCs with an increased Ca2+ permeability were eliminated by phagocytic cells in the spleen, it was found that such RBCs were taken up by marginal zone macrophages and dendritic cells (DCs) in a manner distinct from that of naturally senescent RBCs. The DC population particularly efficient in this process expressed CD207 and are known for their ability to promote immunological tolerance. Eryptotic cell uptake was not regulated by the phagocytosis-inhibitory protein CD47 on the RBCs.

To investigate how RBCs damaged during liquid storage are recognized and taken up by macrophages, a model to store and transfuse murine RBCs was developed. This storage model generated murine RBCs with several characteristics similar to that of stored human RBCs (i.e. loss of ATP, formation of RBC microparticles and rapid clearance of up to 35% of the RBCs during the first 24 h after transfusion). In vitro phagocytosis of human as well as murine stored RBCs was serum dependent and could be inhibited by blocking class A scavenger receptors using fucoidan or dextran sulphate.

In conclusion, the findings of this thesis contribute to further understanding how changes inflicted to RBCs during storage direct the fate of these cells in their interaction with cells of the immune system after transfusion. The observation of an increased Ca2+ permeability of stored RBCs, and the possible recognition of such cells by tolerance-promoting DCs, in combination with the findings that class A scavenger receptors and serum factors may mediate recognition of stored RBCs, may result in novel new directions of research within the field of transfusion medicine.

Place, publisher, year, edition, pages
Umeå: Umeå universitet , 2015. , 58 p.
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 1727
Keyword [en]
Red blood cell, erythrocyte, transfusion, macrophage, phagocytosis, storage lesion, eryptosis, Class A scavenger receptor
National Category
Cell and Molecular Biology
Research subject
transfusionsmedicin
Identifiers
URN: urn:nbn:se:umu:diva-103270ISBN: 978-91-7601-288-8 (print)OAI: oai:DiVA.org:umu-103270DiVA: diva2:812619
Public defence
2015-06-11, Sal BiA 201, Biologihuset, Johan Bures väg 12, Umeå, 09:00 (Swedish)
Opponent
Supervisors
Available from: 2015-05-21 Created: 2015-05-19 Last updated: 2015-05-21Bibliographically approved
List of papers
1. Transfusion of cryopreserved human red blood cells into healthy humans is associated with rapid extravascular hemolysis without a proinflammatory cytokine response
Open this publication in new window or tab >>Transfusion of cryopreserved human red blood cells into healthy humans is associated with rapid extravascular hemolysis without a proinflammatory cytokine response
2013 (English)In: Transfusion, ISSN 0041-1132, E-ISSN 1537-2995, Vol. 53, no 1, 28-33 p.Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Transfusion of stored red blood cells (RBCs) can be associated with adverse side effects. Recent studies in mice transfused with stored RBCs showed that a strong proinflammatory cytokine storm was induced due to extravascular hemolysis already at 2 hours after transfusion. Therefore, we here investigated if transfusion of 2 units of cryopreserved autologous RBCs induced a proinflammatory response in healthy human volunteers.

STUDY DESIGN AND METHODS: Two units of autologous RBCs, cryopreserved for 16 weeks, were transfused into 10 healthy human volunteers. Serum and blood samples taken at 2 hours before and at 2 and 48 hours after transfusion were analyzed for signs of extravascular hemolysis and the presence of proinflammatory cytokines.

RESULTS: At 2 hours after transfusion, transferin-bound serum iron, as well as transferin saturation and total bilirubin, were already significantly increased. These measures all returned back toward that in pretransfusion samples at 48 hours after transfusion. No increases in the production of the proinflammatory cytokines interleukin (IL)-1β, IL-6, IL-8, monocyte chemotactic protein-1, macrophage inflammatory protein-1β, or tumor necrosis factor-α were detected at any time point after transfusion.

CONCLUSION: Although a significant level of extravascular hemolysis already occurred at 2 hours after transfusion of cryopreserved RBCs, there were no signs of proinflammatory cytokine production up to 48 hours after transfusion.

Place, publisher, year, edition, pages
John Wiley & Sons, 2013
National Category
Sport and Fitness Sciences
Identifiers
urn:nbn:se:umu:diva-55643 (URN)10.1111/j.1537-2995.2012.03710.x (DOI)000313348900007 ()22612879 (PubMedID)
Available from: 2012-05-25 Created: 2012-05-25 Last updated: 2017-12-07Bibliographically approved
2. Splenic uptake of RBCs with an elevated cytoplasmic Ca2+-concentration primarily involves marginal zone macrophages and CD207+ dendritic cells
Open this publication in new window or tab >>Splenic uptake of RBCs with an elevated cytoplasmic Ca2+-concentration primarily involves marginal zone macrophages and CD207+ dendritic cells
Show others...
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Normally senescent red blood cells (RBCs) have a rather fixed life-span after which they are eliminated from the circulation mainly by macrophages in the spleen and liver. However, the normal life-span may be significantly reduced when RBCs are stored under blood bank conditions, which ultimately results in a reduced 24 hour survival after transfusion. Although not completely understood, the damage occurring to some of the stored RBCs is probably multifactorial as stored RBCs shares features of both senescence and suicidal RBC death (eryptosis). One key cellular event associated with eryptosis is an increased intracellular Ca2+-concentration. We found that human RBCs stored for up to 42 days under blood bank conditions contained a small subset of cells with an increased intracellular Ca2+-concentration corresponding to that during eryptosis. Since little is known about the mechanisms mediating uptake and clearance of eryptotic RBCs in vivo, we further investigated this using a murine transfusion model of Ca2+ ionophore-treated RBCs (Ca2+-RBCs). Ca2+-RBCs were mainly trapped by MARCO+ splenic marginal zone macrophages and CD11c+ dendritic cells (DCs) already at 1 hour after transfusion. Among DCs, CD11c+ CD207+ DCs in the marginal zone were particularly efficient in mediating uptake of Ca2+-RBCs. Similar to that found in vitro, CD47 on the Ca2+-RBCs did not affect their clearance in vivo. Thus, RBCs with an increased intracellular Ca2+-concentration accumulates during RBC storage, and in a murine model such RBCs are recognized by splenic macrophages and DCs in ways similar to what has been reported for nucleated apoptotic cells.

National Category
Cell and Molecular Biology
Research subject
Immunology
Identifiers
urn:nbn:se:umu:diva-103268 (URN)
Available from: 2015-05-19 Created: 2015-05-19 Last updated: 2015-05-20
3. Phagocytosis of liquid-stored red blood cells in vitro requires serum and macrophage scavenger receptors
Open this publication in new window or tab >>Phagocytosis of liquid-stored red blood cells in vitro requires serum and macrophage scavenger receptors
(English)Manuscript (preprint) (Other academic)
Abstract [en]

BACKGROUND: Red blood cells (RBCs) undergo structural and metabolic changes with prolonged storage, which ultimately may decrease their survival after transfusion. Although the storage-induced damage to RBCs has been rather well described biochemically, little is known about the mechanisms underlying the recognition and rapid clearance of the damaged cells by macrophages.STUDY DESIGN AND METHODS: We here used a murine model for cold (+4°C) RBC storage and transfusion. Phagocytosis of human or murine RBCs, liquid stored for 6-8 weeks or 10-14 days respectively, was investigated in murine peritoneal macrophages.RESULTS: The effects of storage on murine RBCs resembled that described for stored human RBCs with regard to decreased adenosine triphosphate (ATP) levels, accumulation of microparticles during storage, and RBC recovery kinetics after transfusion. Under serum-free conditions, phagocytosis of stored human or murine RBCs was reduced by 70-75%, as compared with that in the presence of heat-inactivated fetal calf serum (FCS). Human serum promoted phagocytosis of stored human RBCs similar to that seen with FCS. By blocking macrophage class A scavenger receptors with fucoidan or dextran sulphate, phagocytosis of human or murine RBCs was reduced by more than 90%. Phagocytosis of stored human RBCs was also sensitive to inhibition by the phosphatidylinositol 3 kinase-inhibitor LY294002, the ERK1/2-inhibitor PD98059, or the p38 MAPK-inhibitor SB203580.CONCLUSIONS: RBCs damaged during liquid storage may be recognized by macrophage class A scavenger receptors and serum-dependent mechanisms. This species-independent recognition mechanism may help to further understand the rapid clearance of stored RBCs shortly after transfusion.

National Category
Cell and Molecular Biology
Research subject
transfusionsmedicin
Identifiers
urn:nbn:se:umu:diva-103266 (URN)
Available from: 2015-05-19 Created: 2015-05-19 Last updated: 2015-05-20

Open Access in DiVA

Doctoral thesis A. Hult 2015(1140 kB)871 downloads
File information
File name FULLTEXT01.pdfFile size 1140 kBChecksum SHA-512
3eb56b642213757499adb795a02c477827f6db57aa1ba4e704b79ecbd9a180abc2133fcccdb28dbd9ebb16a6ef07364d9206ea976abbb87f8af5c2f7cc6bee43
Type fulltextMimetype application/pdf
Spikblad A. Hult 2015(29 kB)48 downloads
File information
File name FULLTEXT02.pdfFile size 29 kBChecksum SHA-512
a099ab83057c1bf58cbc4f4b03d640a4db089661dabb4a77529af9250a4d1453d9e93102618130349a5132dfd4cfe7a21f628edfac6b06aa077d59aaa38211af
Type spikbladMimetype application/pdf

Search in DiVA

By author/editor
Hult, Andreas
By organisation
IdrottsmedicinDepartment of Surgical and Perioperative SciencesDepartment of Integrative Medical Biology (IMB)
Cell and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 919 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

isbn
urn-nbn

Altmetric score

isbn
urn-nbn
Total: 1953 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf