Monocyte-mediated activation of endothelial cells occurs only after binding to extracellular vesicles from red blood cell products, a process mediated by β-integrin

Transfusion ◽  
2016 ◽  
Vol 56 (12) ◽  
pp. 3012-3020 ◽  
Author(s):  
Marleen Straat ◽  
Maike E. van Hezel ◽  
Anita Böing ◽  
Anita Tuip-De Boer ◽  
Nina Weber ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood Cell ◽  
Extracellular Vesicles ◽  
Red Blood Cell

Additive effect of red blood cell rigidity and adherence to endothelial cells in inducing vascular resistance

2008 ◽  
Vol 295 (4) ◽  
pp. H1788-H1793 ◽  
Author(s):  
D. K. Kaul ◽  
A. Koshkaryev ◽  
G. Artmann ◽  
G. Barshtein ◽  
S. Yedgar
Keyword(s):  
Endothelial Cells ◽  
Blood Cell ◽  
Vascular Resistance ◽  
Red Blood Cell ◽  
Untreated Control ◽  
Additional Increase ◽  
Specific Effects ◽  
Induced Changes ◽  
Rbc Deformability

To explore the contribution of red blood cell (RBC) deformability and interaction with endothelial cells (ECs) to circulatory disorders, these RBC properties were modified by treatment with hydrogen peroxide (H2O2), and their effects on vascular resistance were monitored following their infusion into rat mesocecum vasculature. Treatment with 0.5 mM H2O2 increased RBC/EC adherence without significant alteration of RBC deformability. At 5.0 mM H2O2, RBC deformability was considerably reduced, inducing a threefold increase in the number of undeformable cells, whereas RBC/EC adherence was not further affected by the increased H2O2 concentration. This enabled the selective manipulation of RBC adherence and deformability and the testing of their differential effect on vascular resistance. Perfusion of RBCs with enhanced adherence and unchanged deformability (treatment with 0.5 mM H2O2) increased vascular resistance by about 35% compared with untreated control RBCs. Perfusion of 5.0 mM H2O2-treated RBCs, with reduced deformability (without additional increase of adherence), further increased vascular resistance by about 60% compared with untreated control RBCs. These results demonstrate the specific effects of elevated adherence and reduced deformability of oxidized RBCs on vascular resistance. These effects can be additive, depending on the oxidation conditions. The oxidation-induced changes applied in this study are moderate compared with those observed in RBCs in pathological states. Yet, they caused a considerable increase in vascular resistance, thus demonstrating the potency of RBC/EC adherence and RBC deformability in determining resistance to blood flow in vivo.

scholarly journals Assessment of the Effect of Crizanlizumab on Red Blood Cell Adhesion to Endothelial Cells Using a Standardized Endothelium-on-a-Chip Microfluidic Platform

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 2021-2021
Author(s):  
Erdem Kucukal ◽  
Aaron Wolfe ◽  
Ryan Kocevar ◽  
Lalitha V Nayak ◽  
Andreas Bruederle ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Cell Surface ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Stock Options ◽  
Microfluidic Channels ◽  
Blood Samples ◽  
Privately Held

Abstract Background: Chronic upregulation of P-selectin (P-sel) on blood cells and the endothelium leads to abnormal red blood cell (RBC) adhesion to endothelial cells, significantly contributing to vaso-occlusive crises (VOCs), which are a major cause of morbidity and mortality in patients with sickle cell disease (SCD). Crizanlizumab (criz, a.k.a. SEG101) is a humanized anti-P-sel monoclonal antibody and has recently been approved by the Food and Drug Administration to reduce the frequency of VOCs in SCD patients. Here, we report in vitro assessment of the effect of criz on patient-specific RBC adhesion to heme-activated human endothelial cells using a standardized endothelialized microfluidic platform, the Endothelium-on-a-chip. Methods: Whole blood samples were collected from 13 subjects with SCD (13 HbSS and 1 HbSC) in EDTA vacutainers. RBCs were isolated via centrifugation from whole blood and then resuspended in basal cell culture medium (EBM, Lonza, Morristown, USA) at a hematocrit of 20% buffered with 10 mM of HEPES. Human umbilical vein endothelial cells (HUVECs) were obtained from Lonza and cultured within the microfluidic channels at 15 dyne/cm 2 for at least 48 hours prior to experiments. For long-term activation, HUVECs were treated with 40 µM heme for 4 hours +/- 100 µg/ml criz for 1 hour followed by injection of blood samples through the microfluidic channels. For short-term activation, blood samples were supplemented with 40 µM heme +/- 100 µg/ml criz and injected through the microfluidic channels for 15 minutes. Thereafter, non-adherent RBCs were rinsed via either only heme-containing EGM or heme- and criz-containing EGM, and the remaining RBCs were quantified based on published methods [1]. Student's t-test was used to calculate statistical significance. Results: We found that 4-hour heme activation of HUVECs resulted in significantly elevated RBC adhesion compared to baseline although adhesion levels were heterogenous among the patient population (Fig. 1A, 1671±522 vs 17±4, p<0.05). Treatment of 4-hour heme-activated HUVECs with criz did not significantly decrease RBC adhesion (Fig. 1A, 1170±413 vs 1671±522, p>0.05), while we observed lower RBC adhesion to criz treated HUVECs for certain subjects (Fig. 1B). By contrast, criz treatment significantly reduced the number of adherent RBCs to 15-min heme-activated HUVECs (Fig. 1C, 135±40 vs 1513±617, p<0.05). Next, we assessed whether criz would disrupt already established adhesive interactions between RBCs and 15-min heme-activated HUVECs. To do so, we first allowed RBCs to adhere to heme-activated HUVECs (for 15-min) and then rinsed the microchannels (at 10 μl/min) via either a heme- or both heme- and criz-containing solution (for 15 min). We then quantified the number of adherent RBCs at min=0 and min=15. While only 10% of the adherent RBCs remained in the microchannels following a 15-minute wash with criz, this ratio was 60% without criz (Fig. 1E). Discussion: Our results show that the magnitude of inhibition of RBC adhesion to HUVECs with criz correlated with the duration of heme-activation (4 hours vs 15 minutes). This is likely due to variable levels of different adhesion molecules on acute or chronically activated HUVECs. For instance, it has been shown that P-selectin is rapidly translocated to the cell surface following heme activation [1], but its concentration on cell surface significantly decays with time. Previous experiments have shown that sickle RBCs can adhere to cell adhesion molecules such as ICAM-1 [2], which mechanistically may play a role in the case of a chronically activated endothelium. We are currently exploring whether criz would also reduce RBC adhesion to acutely activated endothelial cells that are under chronic stress. These preliminary results suggest that the Endothelium-on-a-chip, as partner in novel therapeutic studies, could help monitoring dynamics of targeted therapies in SCD patients during drug development and in clinical trials. Acknowledgements: This work was funded by Novartis. The authors would like to thank the Ohio Third Frontier Technology Validation and Start-up Fund (TVSF) and National Science Foundation Phase-I Small Business Technology Transfer (STTR) award, which supported this work in part. Crizanlizumab was donated by Novartis. References: 1. Kucukal, E., et al., American Journal of Hematology, 2018. 93(8): p.1050-60 2. Kucukal, E., et al., Blood Advances, 2020. 4(15):3688-98 Figure 1 Figure 1. Disclosures Kucukal: BioChip Labs: Current Employment, Patents & Royalties. Kocevar: BioChip Labs: Current Employment. Nayak: BioChip Labs: Current Employment. Bruederle: Novartis Pharma AG: Current Employment. Zak: XaTek: Current Employment, Current holder of stock options in a privately-held company; BioChip Labs: Current Employment, Current holder of stock options in a privately-held company; TecTraum Inc: Current Employment, Current holder of stock options in a privately-held company. Gurkan: Dx Now Inc.: Patents & Royalties; Hemex Health, Inc.: Current Employment, Patents & Royalties; Biochip Labs: Patents & Royalties; Xatek Inc.: Patents & Royalties.

scholarly journals Red blood cell-derived extracellular vesicles mediate intercellular communication in ischemic heart failure

2019 ◽  
Author(s):  
Avash Das ◽  
Nedyalka Valkov ◽  
Ane M. Salvador ◽  
Ivan Kur ◽  
Olivia Ziegler ◽  
...  
Keyword(s):  
Heart Failure ◽  
Mouse Model ◽  
Blood Cell ◽  
Extracellular Vesicles ◽  
Red Blood Cell ◽  
Significant Proportion ◽  
Ischemic Heart ◽  
Target Cells ◽  
Ischemic Heart Failure ◽  
Cellular Markers

SummaryExtracellular vesicles (EV) mediate intercellular signaling by transferring their cargo to recipient cells. Red blood cell (RBC)-derived EVs constitute a significant proportion of circulating EVs and have been implicated in regulating immune responses. Here, we describe a transgenic mouse model for fluorescent-based mapping of RBC-EV target cells based on the functional transfer of EV-contained Cre-recombinase to target cells. In a murine model of ischemic heart failure, we detect an increase in RBC-EV-targeted cardiomyocytes in the hearts and microglial cells in the brains. Cells targeted by RBC-EVs present an enrichment of genes implicated in cell proliferation and metabolism pathways compared to non-recombined (non-targeted) cells. Cardiomyocytes targeted by RBC-EVs are more likely to demonstrate cellular markers of DNA synthesis and proliferation, suggesting functional significance of EV-mediated signaling. In conclusion, we leverage our mouse model for mapping of RBC-EV targets in murine ischemic heart failure to demonstrate quantitative and qualitative changes in RBC-EV recipients.

Clearance and phenotype of extracellular vesicles after red blood cell transfusion in a human endotoxemia model

2019 ◽  
Vol 58 (4) ◽  
pp. 508-511 ◽  
Author(s):  
Lisa van Manen ◽  
Anna L. Peters ◽  
P. Matthijs van der Sluijs ◽  
Rienk Nieuwland ◽  
Robin van Bruggen ◽  
...  
Keyword(s):  
Blood Cell ◽  
Extracellular Vesicles ◽  
Red Blood Cell ◽  
Red Blood Cell Transfusion ◽  
Human Endotoxemia ◽  
Endotoxemia Model

Determination of red blood cell adhesion to vascular endothelial cells: A critical role for blood plasma

2021 ◽  
pp. 112226
Author(s):  
Gregory Barshtein ◽  
Orly Zelig ◽  
Alexander Gural ◽  
Dan Arbell ◽  
Saul Yedgar
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Blood Cell ◽  
Blood Plasma ◽  
Red Blood Cell ◽  
Vascular Endothelial Cells ◽  
Critical Role ◽  
Vascular Endothelial

scholarly journals Syngeneic red blood cell–induced extracellular vesicles suppress delayed‐type hypersensitivity to self‐antigens in mice

2019 ◽  
Vol 49 (11) ◽  
pp. 1487-1499 ◽  
Author(s):  
Katarzyna Nazimek ◽  
Eugenio Bustos‐Morán ◽  
Noelia Blas‐Rus ◽  
Bernadeta Nowak ◽  
Włodzimierz Ptak ◽  
...  
Keyword(s):  
Blood Cell ◽  
Extracellular Vesicles ◽  
Red Blood Cell ◽  
Delayed Type Hypersensitivity ◽  
Self Antigens

scholarly journals A role for activated endothelial cells in red blood cell clearance: implications for vasopathology

Haematologica ◽  
2011 ◽  
Vol 97 (4) ◽  
pp. 500-508 ◽  
Author(s):  
M. H. A. M. Fens ◽  
R. van Wijk ◽  
G. Andringa ◽  
K. L. van Rooijen ◽  
H. M. Dijstelbloem ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Cell Clearance

scholarly journals The Fluid Membrane determines Mechanics of Red Blood Cell Extracellular Vesicles and is Softened in Hereditary Spherocytosis

2017 ◽  
Author(s):  
Daan Vorselen ◽  
Susan M. van Dommelen ◽  
Raya Sorkin ◽  
Jürgen Schiller ◽  
Richard van Wijk ◽  
...  
Keyword(s):  
Blood Cell ◽  
Extracellular Vesicles ◽  
Red Blood Cell ◽  
Hereditary Spherocytosis ◽  
Cell Communication ◽  
Lipid Vesicle ◽  
Bending Modulus ◽  
Rbc Membrane ◽  
Blood Disorder ◽  
Healthy Donors

AbstractExtracellular vesicles (EVs) are widely studied regarding their role in cell-to-cell communication and disease, as well as for applications as biomarker or drug delivery vehicle. EVs contain both membrane and intraluminal proteins, affecting their structural properties and thereby likely their functioning. Here, we use atomic force microscopy for the mechanical characterization of red blood cell (RBC) EVs from healthy individuals as well as from a patient with hereditary spherocytosis (HS) due to ankyrin deficiency. We show that the EVs are packed with proteins, yet their response to indentation is similar to that of a fluid lipid vesicle lacking proteins. The bending modulus of RBC EVs of healthy donors is ~15kbT, agreeing well with the bending modulus of the RBC membrane. Surprisingly, whereas RBCs become more rigid in HS, the excreted vesicles of a patient with this blood disorder have a significantly (~50%) lower bending modulus than donor EVs. These results shed new light on the mechanism and effects of EV budding and may underlie the reported increase in vesiculation and stiffening of RBCs in hereditary spherocytosis patients.

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