Ligation of CD47 mediates phosphatidylserine expression on erythrocytes and a concomitant loss of viability <i>in vitro</i>

Head, David J.; Lee, Zoe E.; Swallah, Muhammed M.; Avent, Neil D.

doi:10.1111/j.1365-2141.2005.05668.x

Cited by 45 publications

(45 citation statements)

References 11 publications

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“…Ligation of GPC markedly increased PS exposure in the control, as previously recorded, 33 and did so, but to a lesser extent, in patients A and B (still less in patient B than in patient A) ( Figure 1). Ligation of CD47 markedly increased PS exposure in the control, as previously observed, 32 and did so to an even greater extent in patients A and B (Figure 1). Ligation of CD44 and GPA produced no particular effect with regard to the control (except that patient B's red cells, as already pointed out, showed spontaneously increased PS exposure) (Figure 1).…”

Section: Receptor-mediated Exposure Of Phosphatidylserine In 41r (-)supporting

confidence: 85%

“…Ligation of CD47 using monoclonal antibody BRIC 126 and 4N1K peptidemediated PS exposure on red cells is associated with a loss of viability in vitro. 32 Ligation of GPC (with mouse monoclonal antibody BRIC-10) also caused PS exposure and demonstrated similar effects. Notably though, this effect was cancelled with mutant forms of GPC, missing exon 2 (Yus phenotype) or exon 3 (Gerbich phenotype) in the GYPC gene, 33 but was unchanged with elongated GPC variant Ls a (duplication of exon 3).…”

Section: Introductionmentioning

confidence: 87%

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4.1R-deficient human red blood cells have altered phosphatidylserine exposure pathways and are deficient in CD44 and CD47 glycoproteins

Jeremy¹,

Plummer²,

Head³

et al. 2009

Haematologica

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BackgroundProtein 4.1R is an important component of the red cell membrane skeleton. It imparts structural integrity and has transmembrane signaling roles by direct interactions with transmembrane proteins and other membrane skeletal components, notably p55 and calmodulin. Design and MethodsSpontaneous and ligation-induced phosphatidylserine exposure on erythrocytes from two patients with 4.1R deficiency were studied, using CD47 glycoprotein and glycophorin C as ligands. We also looked for protein abnormalities in the 4.1R -based multiprotein complex. ResultsPhosphatidylserine exposure was significantly increased in 4.1R-deficient erythrocytes obtained from the two different individuals when ligands to CD47 glycoprotein were bound. Spontaneous phosphatidylserine exposure was normal. 4.1R, glycophorin C and p55 were missing or sharply reduced. Furthermore there was an alteration or deficiency of CD47 glycoprotein and a lack of CD44 glycoprotein. Based on a recent study in 4.1R-deficient mice, we found that there are clear functional differences between interactions of human red cell 4.1R and its murine counterpart. ConclusionsGlycophorin C is known to bind 4.1R, and we have defined previously that it also binds CD47. From our evidence, we suggest that 4.1R plays a role in the phosphatidylserine exposure signaling pathway that is of fundamental importance in red cell turnover. The linkage of CD44 to 4.1R may be relevant to this process. CD44 and CD47 glycoproteins. Haematologica 2009;94:1354-1361. doi:10.3324/haematol.2009 This is an open-access paper. 4.1R-deficient human red blood cells have altered phosphatidylserine exposure pathways and are deficient in CD44 and CD47 glycoproteins

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Section: Receptor-mediated Exposure Of Phosphatidylserine In 41r (-)supporting

confidence: 85%

Section: Introductionmentioning

confidence: 87%

4.1R-deficient human red blood cells have altered phosphatidylserine exposure pathways and are deficient in CD44 and CD47 glycoproteins

Jeremy¹,

Plummer²,

Head³

et al. 2009

Haematologica

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“…Intriguingly, TSP-1 has also been shown to be able to induce erythrocyte phosphatidylserine exposure through CD47 after prolonged incubation. 38 We therefore propose a model in which the conformational change in CD47 and subsequent TSP-1 binding are at the basis of the recognition of aged erythrocytes as well as the generation of additional "eat me" signals How TSP-1 is able to induce phagocytosis of old erythrocytes through binding to CD47 and subsequent recognition by SIRP␣ is currently unknown. Although the results from the phagocytosis of aged erythrocytes treated with the TSP-1 peptide might suggest that TSP-1 is simply blocking the CD47-SIRP␣ interaction, this is not in line with the results from the CHO SIRP␣ binding.…”

Section: Discussionmentioning

confidence: 99%

“…28,29,36,37 Interestingly, TSP-1 binding to CD47 on erythrocytes induces erythrocyte death, a response that can be mimicked by a C-terminal peptide derived from TSP-1, 4N1K. 38 Moreover, changes in CD47 conformation on sickle cell disease erythrocytes induce binding of TSP-1 to CD47. 35 Control and oxidized erythrocytes were incubated with either biotinylated 4N1K peptide or a biotinylated control peptide, 4NGG.…”

Section: Cd47 On Experimentally Aged Erythrocytes Is Recognized By Sirp␣mentioning

confidence: 99%

CD47 functions as a molecular switch for erythrocyte phagocytosis

Burger

Hilarius-Stokman

Korte

et al. 2012

Blood

276

289

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CD47 on erythrocytes inhibits phagocytosis through interaction with the inhibitory immunoreceptor SIRP␣ expressed by macrophages. Thus, the CD47-SIRP␣ interaction constitutes a negative signal for erythrocyte phagocytosis. However, we report here that CD47 does not only function as a "do not eat me" signal for uptake but can also act as an "eat me" signal. In particular, a subset of old erythrocytes present in whole blood was shown to bind and to be phagocytosed via CD47-SIRP␣ interactions. Furthermore, we provide evidence that experimental aging of erythrocytes induces a conformational change in CD47 that switches the molecule from an inhibitory signal into an activating one. Preincubation of experimentally aged erythrocytes with human serum before the binding assay was required for this activation. We also demonstrate that aged erythrocytes have the capacity to bind the CD47-binding partner thrombospondin-1 (TSP-1) and that treatment of aged erythrocytes with a TSP-1-derived peptide enabled their phagocytosis by human red pulp macrophages. Finally, CD47 on erythrocytes that had been stored for prolonged time was shown to undergo a conformational change and bind TSP-1. These findings reveal a more complex role for CD47-SIRP␣ interactions in erythrocyte phagocytosis, with CD47 acting as a molecular switch for controlling erythrocyte phagocytosis. (Blood. 2012;119(23):5512-5521) IntroductionErythrocyte clearance has been studied for many years in the context of normal turnover, 1,2 enhanced clearance resulting from defects in erythrocyte metabolism or changes in membrane composition, [3][4][5] and in blood transfusion. 6 A large number of factors that may influence erythrocyte clearance have been proposed, ranging from autoantibody binding to band 3 after conformational changes resulting from aging (the "senescent cell antigen"), 7,8 to the expression of phosphatidylserine on the outer leaflet of the erythrocyte membrane. 9 In general, it is thought that erythrocytes are cleared after accumulating so-called "eat me" signals and are then phagocytosed by macrophages of the reticuloendothelial system, predominantly in the spleen and liver. 1,10,11 In contrast, one of the membrane proteins expressed by erythrocytes, CD47, has been shown to inhibit phagocytosis of erythrocytes by macrophages of the reticuloendothelial system. 12-16 CD47 exerts its inhibitory effect through binding to SIRP␣ on the macrophage, which induces inhibitory signaling by the immunoreceptor tyrosinebased inhibition motifs (ITIMs) residing in the cytoplasmic tail of SIRP␣. 17,18 On ligation of SIRP␣ by CD47, the SIRP␣ ITIMs recruit and activate tyrosine phosphatases SHP-1 and SHP-2, and this regulates, generally in a negative fashion, downstream signaling pathways and effector functions. The inhibitory effect of CD47 on erythrocyte clearance can be illustrated by transfusion of CD47-deficient erythrocytes into a wild-type recipient, which leads to rapid phagocytosis of the CD47-deficient erythrocytes by red pulp macrophages in the spleen. 12,19 ...

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“…RBC ligands, even monovalent, specifically targeted to GPA and band 3, have the potential to cause undesirable alterations of RBC, including changes in deformability, [24][25][26][27][28] exposure of phosphatidylserine (PS), 29 and generation of reactive oxygen species (ROS). 30 Although antibodies to GPA and band 3 generally reduce membrane deformability, in early studies of the membrane deformability of antibody-coated RBCs, others observed that for some antigens, nonspherocytic antibody-sensitized RBCs could maintain normal deformability.…”

Section: Introductionmentioning

confidence: 99%

Biocompatible coupling of therapeutic fusion proteins to human erythrocytes

et al. 2018

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Key Points• Thrombomodulin was fused to scFvs targeting RhCE (Rh17 epitope) and band 3/GPA (Wr b epitope).• Fusion proteins were efficacious in a humanized microfluidic model of inflammatory thrombosis. fibrin deposition induced by tumor necrosis factor-a in an endothelialized microfluidic model using human whole blood. RhCE-bound hTM-scFv more effectively reduced platelet and leukocyte adhesion, whereas anti-Wr b scFv appeared to promote platelet adhesion.These data provide a translational framework for the development of engineered affinity ligands to safely couple therapeutics to human RBCs.

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Ligation of CD47 mediates phosphatidylserine expression on erythrocytes and a concomitant loss of viability in vitro

Cited by 45 publications

References 11 publications

4.1R-deficient human red blood cells have altered phosphatidylserine exposure pathways and are deficient in CD44 and CD47 glycoproteins

4.1R-deficient human red blood cells have altered phosphatidylserine exposure pathways and are deficient in CD44 and CD47 glycoproteins

CD47 functions as a molecular switch for erythrocyte phagocytosis

Biocompatible coupling of therapeutic fusion proteins to human erythrocytes

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