scholarly journals Differential Expression of CD31 and Von Willebrand Factor on Endothelial Cells in Different Regions of the Human Brain: Potential Implications for Cerebral Malaria Pathogenesis

2020 ◽  
Vol 10 (1) ◽  
pp. 31 ◽  
Author(s):  
Smart Ikechukwu Mbagwu ◽  
Luis Filgueira
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Human Brain ◽  
Expression Pattern ◽  
Von Willebrand Factor ◽  
Brain Regions ◽  
And Function ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
The Brain

Cerebral microvascular endothelial cells (CMVECs) line the vascular system of the brain and are the chief cells in the formation and function of the blood brain barrier (BBB). These cells are heterogeneous along the cerebral vasculature and any dysfunctional state in these cells can result in a local loss of function of the BBB in any region of the brain. There is currently no report on the distribution and variation of the CMVECs in different brain regions in humans. This study investigated microcirculation in the adult human brain by the characterization of the expression pattern of brain endothelial cell markers in different brain regions. Five different brain regions consisting of the visual cortex, the hippocampus, the precentral gyrus, the postcentral gyrus, and the rhinal cortex obtained from three normal adult human brain specimens were studied and analyzed for the expression of the endothelial cell markers: cluster of differentiation 31 (CD31) and von-Willebrand-Factor (vWF) through immunohistochemistry. We observed differences in the expression pattern of CD31 and vWF between the gray matter and the white matter in the brain regions. Furthermore, there were also regional variations in the pattern of expression of the endothelial cell biomarkers. Thus, this suggests differences in the nature of vascularization in various regions of the human brain. These observations also suggest the existence of variation in structure and function of different brain regions, which could reflect in the pathophysiological outcomes in a diseased state.

PERTURBATION OF CULTURED ENDOTHELIAL CELLS ALTERS CELLULAR VON WILLEBRAND FACTOR DISTRIBUTION

1987 ◽  
Author(s):  
J H Reinders ◽  
C L Verweii ◽  
J A V Mourlk ◽  
Ph G de Groot
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Phorbol Esters ◽  
Term Treatment ◽  
Cellular Distribution ◽  
Long Term Treatment ◽  
Von Willebrand ◽  
Willebrand Factor

Endothelial cells, cultured from human umbilical veins, synthesize von Willebrand Factor (vWF), that is stored by the cells in Weibel-Palade bodies, secreted into the medium and incorporated into the extracellular matrix underneath the cells. We have studied the influence of perturbation by phorbol esters and thrombin on the cellular distribution of vWF. Short-term (< 1 hour) treatment of endothelial cells with phorbol ester PMA or thrombin resulted in the release of cellular stored vWF. Long-term treatment with perturbants evoked a distinct change in the endothelial cell distribution of vWF, evident 24 to 48 hours after exposure. While the contents of the vWF storage vesicles were gradually restored within 48 hours, enhanced amounts of vWF were secreted into the medium. However, PMA did not increase the endothelial cell contents of mRNA encoding for vWF. The number as well as the size of vWF storage granules in the cells increased after exposure to perturbants. The perturbed cells responded to stimuli in releasing stored vWF, the amounts secreted were even greater than those in control cells. The extracellular matrix lost its vWF contents as the result of PMA or thrombin treatment, by blocking deposition of vWF in the matrix, not by enhancing degradation of matrix vWF. In perfusion experiments, the adhesion of washed platelets onto the isolated matrix of perturbed cells was considerable less than that in controls. Addition of vWF to the perfusate overcame this impairment. Thus, perturbation of endothelial cells changes the cellular distribution of vWF.Supported in part by ZWO grants 13-30-31 and 13-90-91 and Netherlands Heart Foundation grant 28.004.

Homocysteine inhibits von Willebrand factor processing and secretion by preventing transport from the endoplasmic reticulum

Blood ◽  
1993 ◽  
Vol 81 (3) ◽  
pp. 683-689 ◽  
Author(s):  
SR Lentz ◽  
JE Sadler
Keyword(s):  
Endothelial Cells ◽  
Endoplasmic Reticulum ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Protein Transport ◽  
Cell Protein ◽  
Carboxyl Terminal ◽  
Insulinoma Cells ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract Intracellular protein transport in endothelial cells is selectively inhibited by homocysteine, a thiol amino acid associated with both thrombosis and atherosclerosis. In a previous study, homocysteine decreased cell surface expression of the surface transmembrane glycoprotein thrombomodulin without decreasing secretion of another endothelial cell protein, plasminogen activator inhibitor-1. To define further the effects of homocysteine on protein transport, we examined the processing and secretion of the multimeric glycoprotein von Willebrand factor (vWF) in human umbilical vein endothelial cells. Incubation with 2 mmol/L homocysteine resulted in complete loss of vWF multimers and prevented asparagine-linked oligosaccharide maturation, propeptide cleavage, and secretion; these effects are consistent with impaired exit from the endoplasmic reticulum (ER). Dimerization was only partially inhibited, suggesting that homocysteine causes retention of provWF in the ER without preventing dimer formation. In pulse-chase incubations, intracellular provWF was degraded before exiting the ER in homocysteine-treated cells. Homocysteine also inhibited the processing and secretion of a carboxyl-terminal truncation mutant of human provWF expressed in rat insulinoma cells, indicating that retention in the endoplasmic reticulum can be mediated by regions of provWF apart from the carboxyl-terminal 20-Kd segment. These results suggest that retention of secretory proteins in the ER is regulated by redox mechanisms and imply that the intracellular transport of multiple endothelial cell proteins may be altered in patients with homocystinuria.

scholarly journals The Interaction of the Platelet and von Willebrand Factor

1977 ◽  
Author(s):  
D.N. Fass ◽  
F. Booyse ◽  
J.C. Lewis ◽  
E. J. W. Bowie
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Cultures ◽  
Von Willebrand Factor ◽  
Sandwich Technique ◽  
Aortic Endothelial Cells ◽  
Confluent Culture ◽  
Endothelial Cell Cultures ◽  
Von Willebrand ◽  
Willebrand Factor

A culture of pig aortic endothelial cells was used for experiments to investigate the interaction between the platelet and von Willebrand factor. An antibody was raised in rabbits to purified porcine von Willebrand factor. A semi-confluent culture of pig endothelial cells was stained immunofluorescently by the sandwich technique using anti-Willebrand factor IgG. An extensive extracellular meshwork of microfilaments was revealed. In endothelial cell cultures from von Willebrand pigs, no immunoreactive microfilaments were found. Immunoelectronmicro-scopy with peroxidase linked antibody has been used to identify similar filaments in normal pig endothelial cells. Washed platelets were shown to adhere to semiconfluent or damaged normal endothelial cell cultures. If the cultures had been previously incubated with anti-Willebrand factor IgG, the washed platelets did not adhere. There was no adherence of platelets when they were added to semiconfluent or damaged von Willebrand endothelial cells.

scholarly journals ABO blood group is a determinant of von Willebrand factor protein levels in human pulmonary endothelial cells

2019 ◽  
Vol 73 (6) ◽  
pp. 347-349 ◽  
Author(s):  
Glenn P Murray ◽  
Steven R Post ◽  
Ginell R Post
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Blood Group ◽  
Von Willebrand Factor ◽  
Blood Type ◽  
Abo Blood Group ◽  
Blood Group Antigens ◽  
Protein Levels ◽  
Von Willebrand ◽  
Willebrand Factor

ABO blood group antigens are expressed on von Willebrand factor (VWF) and glycosylation patterns influence circulating VWF levels. The aim of this study was to examine the effect of ABO blood type on tissue-associated VWF protein levels. We selected 35 formalin-fixed paraffin-embedded pulmonary tissue blocks obtained at autopsy from decedents who died from pulmonary embolism with known ABO blood groups (O, A, B and AB phenotypes), prepared tissue microarrays (TMAs) and stained TMAs with antibodies to VWF and platelet/endothelial cell adhesion marker-1 (PECAM-1) as a marker of endothelial cells. A pixel count scoring algorithm was used to quantify VWF and PECAM-1 staining intensity in pulmonary arterioles in digitised images. Compared with type O, non-O individuals have a significantly higher amount of endothelial cell-associated VWF protein expression. VWF protein levels associated with pulmonary vascular endothelial cells is influenced by ABO antigenic determinants.

Reduction of von Willebrand Factor by Endothelial Cells

2000 ◽  
Vol 84 (09) ◽  
pp. 506-513 ◽  
Author(s):  
Lijuan Xie ◽  
Colin Chesterman ◽  
Philip Hogg
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Protein Disulfide Isomerase ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Reducing Activity ◽  
Von Willebrand ◽  
Protein Disulfide ◽  
Willebrand Factor

SummaryThe haemostatic activity of plasma von Willebrand factor (vWF) is a function of multimer size. Only the large vWF multimers are effective in promoting platelet adhesion to a site of vascular injury. We observed that the conditioned medium of cultured human umbilical vein, human microvascular and bovine aortic endothelial cells contained an activity which reduced the average multimer size of plasma or purified vWF. The average multimer size of vWF produced endogenously by human umbilical vein endothelial cells was similarly reduced following secretion. The reducing activity was ablated by pre-treatment with heat or the thiol blocking agents, iodoacetamide, N-ethylmaleimide or E-64, but not by a range of specific serine-, cysteine-, aspartic-, or metalloproteinase inhibitors. Reduction in vWF multimer size was associated with formation of new thiols in vWF and there was no evidence for additional proteolytic processing of vWF. The reducing activity was associated with a protein with an anionic pI that binds heparin and contains reactive thiol(s). These results suggested that the interchain disulfide bonds that link the vWF homodimers near the N-termini were being reduced by a vWF reductase secreted by endothelial cells. In support of this hypothesis, incubation of vWF with the protein reductants, protein disulfide isomerase and thioredoxin, resulted in formation of new thiols in vWF and reduction in the average multimer size of vWF. These findings may have consequences for control of vWF haemostatic activity. Abbreviations: BAEC, bovine aortic endothelial cell; BSA, bovine serum albumin; cm, conditioned medium; GSH, reduced glutathione; HUVEC, human umbilical vein endothelial cell; HDMVEC, human dermal microvascular endothelial cell; IAM, iodoacetamide; MPB, 3-(N-maleimidylpropionyl)biocytin; NEM, N-ethylmaleimide; PDI, protein disulfide isomerase; PVDF, polyvinylidene difluoride; vWF, von Willebrand factor.

scholarly journals Relative Importance of the Glycoprotein Ib-Binding Domain and the RGD Sequence of von Willebrand Factor for Its Interaction With Endothelial Cells

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2335-2344 ◽  
Author(s):  
Christelle Perrault ◽  
Hanneke Lankhof ◽  
Dominique Pidard ◽  
Danièle Kerbiriou-Nabias ◽  
Jan J. Sixma ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Binding Domain ◽  
Rgd Sequence ◽  
Cultured Endothelial Cells ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Endothelial Cell Adhesion

Endothelial cell adhesion to von Willebrand Factor is mainly mediated through an interaction between the αvβ3 integrin and the RGD sequence of von Willebrand factor (vWF ). To define the potential involvement of glycoprotein Ibα (GPIbα) as an endothelial vWF receptor, we compared cell adhesion to three recombinant vWF, the wild-type (WT-rvWF ) and two mutants, RGGS-rvWF (D1746G), defective for binding to platelet αIIbβ3, and ΔA1-rvWF with a deletion between amino-acids 478 and 716, which does not bind to platelet GPIbα. Adhesion of human umbilical vein endothelial cells to purified vWF recombinants was measured by automatized cell counting using an image analyzer. Whereas cell adhesion to ΔA1-rvWF was unchanged compared with WT-rvWF, reaching a plateau of 40% total cells at a concentration of 2.5 μg/mL rvWF, adhesion to RGGS-rvWF was only 10% of total cells. Cell stimulation by tumor necrosis factor-α (TNFα), reported to upregulate the expression of the putative endothelial GPIbα, did not modify adhesion to these rvWF. Monoclonal antibodies to vWF or GPIbα, blocking vWF interaction with platelet GPIbα, were unable to inhibit endothelial cell adhesion to rvWF. In contrast, antibody 9 to vWF, blocking the αvβ3-dependent endothelial cell adhesion to plasma vWF, inhibited adhesion to WT-rvWF as efficiently as to ΔA1-rvWF (50% inhibition at a concentration of 11 and 15 μg/mL, respectively). In agreement with the fact that endothelial cell adhesion to vWF appeared independent of the GPIbα-binding domain, we were unable to detect endothelial surface expression of GPIbα by flow cytometry or in cell lysates by immunoprecipitation followed by immunoblotting. Moreover, expression of GPIbα mRNA was undetectable in endothelial cells, even after stimulation by TNFα. These studies indicate that GPIbα is not expressed in human cultured endothelial cells and is not involved in adhesion to vWF-containing surfaces. Thus, in static conditions, cultured endothelial cells adhere to vWF through an αvβ3-dependent, GPIbα-independent mechanism.

scholarly journals Homocysteine inhibits von Willebrand factor processing and secretion by preventing transport from the endoplasmic reticulum

Blood ◽  
1993 ◽  
Vol 81 (3) ◽  
pp. 683-689 ◽  
Author(s):  
SR Lentz ◽  
JE Sadler
Keyword(s):  
Endothelial Cells ◽  
Endoplasmic Reticulum ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Protein Transport ◽  
Cell Protein ◽  
Carboxyl Terminal ◽  
Insulinoma Cells ◽  
Von Willebrand ◽  
Willebrand Factor

Intracellular protein transport in endothelial cells is selectively inhibited by homocysteine, a thiol amino acid associated with both thrombosis and atherosclerosis. In a previous study, homocysteine decreased cell surface expression of the surface transmembrane glycoprotein thrombomodulin without decreasing secretion of another endothelial cell protein, plasminogen activator inhibitor-1. To define further the effects of homocysteine on protein transport, we examined the processing and secretion of the multimeric glycoprotein von Willebrand factor (vWF) in human umbilical vein endothelial cells. Incubation with 2 mmol/L homocysteine resulted in complete loss of vWF multimers and prevented asparagine-linked oligosaccharide maturation, propeptide cleavage, and secretion; these effects are consistent with impaired exit from the endoplasmic reticulum (ER). Dimerization was only partially inhibited, suggesting that homocysteine causes retention of provWF in the ER without preventing dimer formation. In pulse-chase incubations, intracellular provWF was degraded before exiting the ER in homocysteine-treated cells. Homocysteine also inhibited the processing and secretion of a carboxyl-terminal truncation mutant of human provWF expressed in rat insulinoma cells, indicating that retention in the endoplasmic reticulum can be mediated by regions of provWF apart from the carboxyl-terminal 20-Kd segment. These results suggest that retention of secretory proteins in the ER is regulated by redox mechanisms and imply that the intracellular transport of multiple endothelial cell proteins may be altered in patients with homocystinuria.

scholarly journals Relative Importance of the Glycoprotein Ib-Binding Domain and the RGD Sequence of von Willebrand Factor for Its Interaction With Endothelial Cells

Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2335-2344 ◽  
Author(s):  
Christelle Perrault ◽  
Hanneke Lankhof ◽  
Dominique Pidard ◽  
Danièle Kerbiriou-Nabias ◽  
Jan J. Sixma ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Binding Domain ◽  
Rgd Sequence ◽  
Cultured Endothelial Cells ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Endothelial Cell Adhesion

AbstractEndothelial cell adhesion to von Willebrand Factor is mainly mediated through an interaction between the αvβ3 integrin and the RGD sequence of von Willebrand factor (vWF ). To define the potential involvement of glycoprotein Ibα (GPIbα) as an endothelial vWF receptor, we compared cell adhesion to three recombinant vWF, the wild-type (WT-rvWF ) and two mutants, RGGS-rvWF (D1746G), defective for binding to platelet αIIbβ3, and ΔA1-rvWF with a deletion between amino-acids 478 and 716, which does not bind to platelet GPIbα. Adhesion of human umbilical vein endothelial cells to purified vWF recombinants was measured by automatized cell counting using an image analyzer. Whereas cell adhesion to ΔA1-rvWF was unchanged compared with WT-rvWF, reaching a plateau of 40% total cells at a concentration of 2.5 μg/mL rvWF, adhesion to RGGS-rvWF was only 10% of total cells. Cell stimulation by tumor necrosis factor-α (TNFα), reported to upregulate the expression of the putative endothelial GPIbα, did not modify adhesion to these rvWF. Monoclonal antibodies to vWF or GPIbα, blocking vWF interaction with platelet GPIbα, were unable to inhibit endothelial cell adhesion to rvWF. In contrast, antibody 9 to vWF, blocking the αvβ3-dependent endothelial cell adhesion to plasma vWF, inhibited adhesion to WT-rvWF as efficiently as to ΔA1-rvWF (50% inhibition at a concentration of 11 and 15 μg/mL, respectively). In agreement with the fact that endothelial cell adhesion to vWF appeared independent of the GPIbα-binding domain, we were unable to detect endothelial surface expression of GPIbα by flow cytometry or in cell lysates by immunoprecipitation followed by immunoblotting. Moreover, expression of GPIbα mRNA was undetectable in endothelial cells, even after stimulation by TNFα. These studies indicate that GPIbα is not expressed in human cultured endothelial cells and is not involved in adhesion to vWF-containing surfaces. Thus, in static conditions, cultured endothelial cells adhere to vWF through an αvβ3-dependent, GPIbα-independent mechanism.

scholarly journals Ionizing irradiation increases transcription of the von Willebrand factor gene in endothelial cells

Blood ◽  
1996 ◽  
Vol 88 (10) ◽  
pp. 3801-3814 ◽  
Author(s):  
N Jahroudi ◽  
AM Ardekani ◽  
JS Greenberger
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Promoter Activity ◽  
Core Promoter ◽  
Irradiation Damage ◽  
Mrna Levels ◽  
Ionizing Irradiation ◽  
Von Willebrand ◽  
Willebrand Factor

Ionizing irradiation damage to the vasculature results in an increase in procoagulant activity of endothelial cells, including elevated von Willebrand factor (vWf) secretion. We investigated the mechanism of irradiation induction of vWf release and demonstrated that vWf mRNA levels were increased when either human or bovine endothelial cells were exposed to 20 Gy irradiation. This response to irradiation was independent to de novo protein synthesis, but required new transcription. Nuclear run-on experiments indicated that increased vWf transcriptional activity was partly responsible for the higher levels of the mRNA accumulation. Transfection analyses with plasmids in which a human growth hormone structural gene was under the control of the endothelial-cell-specific vWf promoter demonstrated that irradiation increased promoter activity. Deletion analyses demonstrated that sequences necessary for irradiation induction of the promoter activity were located within the 112-bp sequences (-90 to +22) that constitute the non-endothelial-cell-specific core promoter region of the vWf gene. Results of gel mobility assays and deletion analyses demonstrated that a site in the vWf promoter other than the putative NF-kB binding site is involved in the mechanism of irradiation induction of the vWf.

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