scholarly journals Hematopoietic and nonhematopoietic cell tissue factor activates the coagulation cascade in endotoxemic mice

Blood ◽  
2010 ◽  
Vol 116 (5) ◽  
pp. 806-814 ◽  
Author(s):  
Rafal Pawlinski ◽  
Jian-Guo Wang ◽  
A. Phillip Owens ◽  
Julie Williams ◽  
Silvio Antoniak ◽  
...  
Keyword(s):  
Mouse Model ◽  
Tissue Factor ◽  
Myeloid Cells ◽  
Cell Types ◽  
Selective Inhibition ◽  
Coagulation Cascade ◽  
Cell Type ◽  
Relative Contribution ◽  
Cell Tissue ◽  
Tf Gene

Tissue factor (TF) is the primary activator of the coagulation cascade. During endotoxemia, TF expression leads to disseminated intravascular coagulation. However, the relative contribution of TF expression by different cell types to the activation of coagulation has not been defined. In this study, we investigated the effect of either a selective inhibition of TF expression or cell type-specific deletion of the TF gene (F3) on activation of coagulation in a mouse model of endotoxemia. We found that inhibition of TF on either hematopoietic or nonhematopoietic cells reduced plasma thrombin-antithrombin (TAT) levels 8 hours after administration of bacterial lipopolysaccharide (LPS). In addition, plasma TAT levels were significantly reduced in endotoxemic mice lacking the TF gene in either myeloid cells (TFflox/flox,LysMCre mice) or in both endothelial cells (ECs) and hematopoietic cells (TFflox/flox,Tie-2Cre mice). However, deletion of the TF gene in ECs alone had no effect on LPS-induced plasma TAT levels. Similar results were observed in mice lacking TF in vascular smooth muscle cells. Finally, we found that mouse platelets do not express TF pre-mRNA or mRNA. Our data demonstrate that in a mouse model of endotoxemia activation of the coagulation cascade is initiated by TF expressed by myeloid cells and an unidentified nonhematopoietic cell type(s).

The Effect of Grp78 Inhibition on Tissue Factor Gene Expression, Procoagulant Activity, and Factor Xa Generation.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 1924-1924
Author(s):  
Gourab Bhattacharjee ◽  
Jasimuddin Ahamed ◽  
Brian Pedersen ◽  
Amr El-Sheikh ◽  
Cheng Liu ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Surface ◽  
Tissue Factor ◽  
Factor Xa ◽  
Cell Types ◽  
Procoagulant Activity ◽  
Coagulation Cascade ◽  
Clotting Factor ◽  
Tf Gene

Abstract In vivo biopanning with phage displayed peptide libraries has generated a group of peptide probes which bind selectively to the surface of atherosclerotic plaque endothelium. The highest affinity peptide, EKO130, binds to the 78 kDa glucose regulated protein (Grp78). Grp78 has been demonstrated to play a role in numerous pathological processes as well as a possible role in the local cell surface regulation of the coagulation cascade. The goal of this study is to determine the role of Grp78 in coagulation including plasma clotting, factor Xa (Xa) generation, and tissue factor (TF) gene expression. siRNA mediated inhibition of Grp78 results in a marked increase in TF gene expression in bEND.3 endothelial cells and RAW macrophage-like cells. Antibody mediated inhibition of cell surface Grp78 results in increased TF procoagulant activity and TF-dependent Xa generation in both the endothelial and macrophage cell types. These studies are consistent with results from another laboratory demonstrating that Grp78 over-expression inhibits TF mediated initiation and support of the coagulation protease cascade. Thus, our work indicates that Grp78 suppresses TF at both the functional and molecular level by inhibiting both its thrombogenic potential and gene expression.

Alternative Splicing Is Responsible for the Presence of Two Tissue Factor mRNA Species in LPS Stimulated Human Monocytes

1992 ◽  
Vol 67 (02) ◽  
pp. 272-276 ◽  
Author(s):  
C Paul ◽  
E van der Logt ◽  
Pieter H Reitsma ◽  
Rogier M Bertina
Keyword(s):  
Alternative Splicing ◽  
Tissue Factor ◽  
Stop Codon ◽  
Cell Types ◽  
Northern Analysis ◽  
Human Monocytes ◽  
Mrna Species ◽  
Protein Levels ◽  
Intron 1 ◽  
Tf Gene

SummaryAlthough normally absent from the surface of all circulating cell types, tissue factor (TF) can be induced to appear on circulating monocytes by stimulants like bacterial lipopolysaccharide (LPS) and phorbolesters. Northern analysis of RNA isolated from LPS stimulated human monocytes demonstrates the presence of 2.2 kb and 3.1 kb TF mRNA species. The 2.2 kb message codes for the TF protein. As demonstrated by Northern blot analysis with a variety of TF gene probes, the 3.1 kb message arises from an alternative splicing process which fails to remove 955 bp from intron 1. Because of a stop codon in intron 1 no TF protein is produced from the 3.1 kb transcript. This larger transcript should therefore not be taken into account when comparing TF gene transcription and TF protein levels.

Monocyte Tissue Factor Expression Is Enhanced in Women who Smoke and Use Oral Contraceptives

1999 ◽  
Vol 82 (12) ◽  
pp. 1614-1620 ◽  
Author(s):  
Helma-Meta Terhalle ◽  
Ute Zakel ◽  
Ulrich Maus ◽  
Behnoush Parviz ◽  
Harald Tillmanns ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Oral Contraceptives ◽  
Transcriptional Activation ◽  
Contraceptive Use ◽  
Coagulation Cascade ◽  
Blood Monocytes ◽  
Inhibitor Molecule ◽  
Increased Risk ◽  
Menopausal Women ◽  
Tf Gene

SummaryThe association between use of oral contraceptives (OCs) and increased risk of thromboembolic disease has been firmly established. This risk increases when use of OCs is combined with cigarette smoking. The cellular mechanism favoring an hypercoagulable state under these behaviours is not known. Circulating monocytes are potent activators of the coagulation cascade through their ability to synthesize procoagulant tissue factor (TF). In the present study we report that monocyte TF expression is increased in women who use OCs and smoke.We studied monocyte TF expression in 4 groups of healthy pre-menopausal women (n = 15 each): (1) non-smoking OC non-users, (2) nonsmoking current OC users, (3) smoking OC non-users and (4) smoking OC users. TF expression was assessed on both mRNA and protein levels in unstimulated and LPS-stimulated cells. Transcriptional activation of the TF gene was assessed by analysis of the transcription factor NF-κB and its inhibitor molecule IκBα. Monocyte TF generation was significantly higher in OC users than in women who did not use OCs. Enhanced monocyte TF generation was also observed in smoking women when compared to non-smokers. Strongest monocyte TF expression occurred in women with combined smoking and use of OCs. The enhanced TF expression in monocytes from women using OCs or smoking was based on an increased TF gene transcription following activation of NF-κB. Experiments on cultured monocytes/macrophages demonstrated enhanced IκBα degradation in the presence of estradiol, suggesting that a direct hormone effect is responsible for the observed increase in monocyte TF expression.This study demonstrates that use of OCs and smoking is associated with an increase in monocyte TF expression in pre-menopausal women. Aberrant TF expression by blood monocytes may favour intravascular clotting activation in women with OC therapy. The further enhancement of TF activity observed in women who smoke and use OCs may explain the synergistic effect of smoking on risk of thromboembolic events associated with contraceptive use.

scholarly journals The principal eosinophil peroxidase product, HOSCN, is a uniquely potent phagocyte oxidant inducer of endothelial cell tissue factor activity: a potential mechanism for thrombosis in eosinophilic inflammatory states

Blood ◽  
2006 ◽  
Vol 107 (2) ◽  
pp. 558-565 ◽  
Author(s):  
Jian-Guo Wang ◽  
Shawn A. Mahmud ◽  
Jacob A. Thompson ◽  
Jian-Guo Geng ◽  
Nigel S. Key ◽  
...  
Keyword(s):  
Tissue Factor ◽  
Umbilical Vein ◽  
Hypereosinophilic Syndrome ◽  
Eosinophil Peroxidase ◽  
Cell Tissue ◽  
Reactive Oxidant ◽  
Tf Gene ◽  
Umbilical Vein Endothelial Cells ◽  
Kinase Pathway

AbstractIn vivo, bromide (Br–), nitrite (NO2–), and thiocyanate (SCN–) compete for oxidation by eosinophil peroxidase (EPO) and H2O2, yielding, respectively, HOBr, NO2·, and HOSCN. We have recently shown that SCN– is the strongly preferred substrate for EPO in vivo and that HOSCN, in contrast with other EPO-generated oxidants and HOCl, is a relatively weak, cell-permeant, sulfhydryl (SH)–reactive oxidant. We here show that HOSCN is a uniquely potent (up to 100-fold) phagocyte oxidant inducer of tissue factor (TF) activity in human umbilical vein endothelial cells (HUVECs). This induction is attributable to transcriptional up-regulation of TF gene expression dependent upon both activation of the p65/c-Rel TF-κB transcription factor and activity of the ERK1/2 kinase pathway upstream of Egr-1 and was markedly further enhanced in the presence of wortmannin, an inhibitor of the PI3 kinase/Akt pathway. HOSCN also markedly activates the proinflammatory p65/p50 NF-κB pathway. Based on these findings we hypothesize that HOSCN generated by adherent and infiltrating eosinophils may provoke the development of a prothrombotic and proinflammatory endothelial/endocardial phenotype that promotes the pronounced thrombotic diathesis characteristic of the hypereosinophilic syndrome.

scholarly journals Identifying Gene Expression Programs of Cell-type Identity and Cellular Activity with Single-Cell RNA-Seq

2018 ◽  
Author(s):  
Dylan Kotliar ◽  
Adrian Veres ◽  
M. Aurel Nagy ◽  
Shervin Tabrizi ◽  
Eran Hodis ◽  
...  
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Matrix Factorization ◽  
Cell Types ◽  
Rna Seq ◽  
Cell Type ◽  
Relative Contribution ◽  
Neuronal Synapses ◽  
Type Identity ◽  
Brain Organoid

AbstractIdentifying gene expression programs underlying both cell-type identity and cellular activities (e.g. life-cycle processes, responses to environmental cues) is crucial for understanding the organization of cells and tissues. Although single-cell RNA-Seq (scRNA-Seq) can quantify transcripts in individual cells, each cell’s expression profile may be a mixture of both types of programs, making them difficult to disentangle. Here we illustrate and enhance the use of matrix factorization as a solution to this problem. We show with simulations that a method that we call consensus non-negative matrix factorization (cNMF) accurately infers identity and activity programs, including the relative contribution of programs in each cell. Applied to published brain organoid and visual cortex scRNA-Seq datasets, cNMF refines the hierarchy of cell types and identifies both expected (e.g. cell cycle and hypoxia) and intriguing novel activity programs. We propose that one of the novel programs may reflect a neurosecretory phenotype and a second may underlie the formation of neuronal synapses. We make cNMF available to the community and illustrate how this approach can provide key insights into gene expression variation within and between cell types.

O tissue factor, where art thou?

Blood ◽  
2010 ◽  
Vol 116 (5) ◽  
pp. 676-677 ◽  
Author(s):  
John H. McVey
Keyword(s):  
Mouse Model ◽  
Tissue Factor ◽  
Myeloid Cells ◽  
Intravascular Coagulation

In this issue of Blood, Pawlinski and colleagues identify myeloid cells and an unidentified nonhematopoietic cell(s) as the source of TF responsible for intravascular coagulation in a mouse model of endotoxemia, excluding a role for EC, VSMC, and platelet cell TF expression.1

scholarly journals Tissue-factor coagulant activity of cultured human endothelial and smooth muscle cells and fibroblasts

Blood ◽  
1977 ◽  
Vol 50 (3) ◽  
pp. 387-396 ◽  
Author(s):  
JR Maynard ◽  
BE Dreyer ◽  
MB Stemerman ◽  
FA Pitlick
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Tissue Factor ◽  
Time Course ◽  
Peak Level ◽  
Cell Types ◽  
Maximum Activity ◽  
Cell Tissue ◽  
Thromboplastic Activity ◽  
High Level

The tissue-factor (thromboplastic) activity of cultured human endothelial cells and fibroblasts is low at time of transfer into fresh medium but increases 3–10 fold. Endothelial cells reach peak activity (400 U/10(5) cells) 5–8 hr after subculture. Activity in fibroblast cultures peaks (3000–12,000 U/10(5) cells) 7–12 hr after subculture. After attaining maximum activity, endothelial and fibroblast tissue- factor content decreases in a time course similar to other cells studied in this laboratory, approaching basal levels by 24–50 hr after subculture. If medium over fibroblasts is changed every 12 hr, activity can be sustained at the peak level for an additional day but cannot be maintained at a high level indefinitely. The kinetics of expression of smooth muscle cell tissue factor are markedly different from other cell types. There is always a pronounced lag (30 hr or more) before the activity increases, and then, in most cases, there is no subsequent decline in activity even though the cells are not refed or restimulated. The activity of each of these cell types is cryptic but becomes available after freeze-thaw disruption of cells.

scholarly journals Cell Type-Specific Imaging of Calcium Signaling in Arabidopsis thaliana Seedling Roots Using GCaMP3

2020 ◽  
Vol 21 (17) ◽  
pp. 6385
Author(s):  
William Krogman ◽  
J. Alan Sparks ◽  
Elison B. Blancaflor
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Types ◽  
Root Cell ◽  
Cauliflower Mosaic Virus ◽  
Cell Type ◽  
Root Cells ◽  
Cell Tissue ◽  
Seedling Roots ◽  
Cell Type Specific

Cytoplasmic calcium ([Ca2+]cyt) is a well-characterized second messenger in eukaryotic cells. An elevation in [Ca2+]cyt levels is one of the earliest responses in plant cells after exposure to a range of environmental stimuli. Advances in understanding the role of [Ca2+]cyt in plant development has been facilitated by the use of genetically-encoded reporters such as GCaMP. Most of these studies have relied on promoters such as Cauliflower Mosaic Virus (35S) and Ubiquitin10 (UBQ10) to drive expression of GCaMP in all cell/tissue types. Plant organs such as roots consist of various cell types that likely exhibit unique [Ca2+]cyt responses to exogenous and endogenous signals. However, few studies have addressed this question. Here, we introduce a set of Arabidopsis thaliana lines expressing GCaMP3 in five root cell types including the columella, endodermis, cortex, epidermis, and trichoblasts. We found similarities and differences in the [Ca2+]cyt signature among these root cell types when exposed to adenosine tri-phosphate (ATP), glutamate, aluminum, and salt, which are known to trigger [Ca2+]cyt increases in root cells. These cell type-targeted GCaMP3 lines provide a new resource that should enable more in depth studies that address how a particular environmental stimulus is linked to specific root developmental pathways via [Ca2+]cyt.

scholarly journals Cell and tissue type independent age-associated DNA methylation changes are not rare but common

2018 ◽  
Author(s):  
Tianyu Zhu ◽  
Shijie C Zheng ◽  
Dirk S. Paul ◽  
S. Horvath ◽  
Andrew E. Teschendorff
Keyword(s):  
Dna Methylation ◽  
Cell Types ◽  
Tissue Type ◽  
Tissue Cell ◽  
Cell Type ◽  
Tissue Specific ◽  
Cell Tissue ◽  
A Cell ◽  
High Degree ◽  
Do So

AbstractAge-associated DNA methylation changes have been widely reported across many different tissue and cell types. Epigenetic ‘clocks’ that can predict chronological age with a surprisingly high degree of accuracy appear to do so independently of tissue and cell-type, suggesting that a component of epigenetic drift is cell-type independent. However, the relative amount of age-associated DNAm changes that are specific to a cell or tissue type versus the amount that occurs independently of cell or tissue type is unclear and a matter of debate, with a recent study concluding that most epigenetic drift is tissue-specific. Here, we perform a novel comprehensive statistical analysis, including matched multi cell-type and multi-tissue DNA methylation profiles from the same individuals and adjusting for cell-type heterogeneity, demonstrating that a substantial amount of epigenetic drift, possibly over 70%, is shared between significant numbers of different tissue/cell types. We further show that ELOVL2 is not unique and that many other CpG sites, some mapping to genes in the Wnt and glutamate receptor signaling pathways, are altered with age across at least 10 different cell/tissue types. We propose that while most age-associated DNAm changes are shared between cell-types that the putative functional effect is likely to be tissue-specific.

scholarly journals Cell Type- and Exposure-Specific Modulation of CD63/CD81-Positive and Tissue Factor-Positive Extracellular Vesicle Release in response to Respiratory Toxicants

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Frank R. M. Stassen ◽  
Pascalle H. van Eijck ◽  
Paul H. M. Savelkoul ◽  
Emiel F. M. Wouters ◽  
Gernot G. U. Rohde ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cell Surface ◽  
Tissue Factor ◽  
Preventive Treatment ◽  
Cell Types ◽  
Extracellular Vesicle ◽  
Bronchial Epithelial Cells ◽  
Specific Response ◽  
Cell Type ◽  
Bronchial Epithelial

Chronic exposure to respiratory stressors increases the risk for pulmonary and cardiovascular diseases. Previously, we have shown that cigarette smoke extract (CSE) triggers the release of CD63+CD81+ and tissue factor (TF)+ procoagulant extracellular vesicles (EVs) by bronchial epithelial cells via depletion of cell surface thiols. Here, we hypothesized that this represents a universal response for different pulmonary cell types and respiratory exposures. Using bead-based flow cytometry, we found that bronchial epithelial cells and pulmonary fibroblasts, but not pulmonary microvascular endothelial cells or macrophages, release CD63+CD81+ and TF+ EVs in response to CSE. Cell surface thiols decreased in all cell types upon CSE exposure, whereas depletion of cell surface thiols using bacitracin only triggered EV release by epithelial cells and fibroblasts. The thiol-antioxidant NAC prevented the EV induction by CSE in epithelial cells and fibroblasts. Exposure of epithelial cells to occupational silica nanoparticles and particulate matter (PM) from outdoor air pollution also enhanced EV release. Cell surface thiols were mildly decreased and NAC partly prevented the EV induction for PM10, but not for silica and PM2.5. Taken together, induction of procoagulant EVs is a cell type-specific response to CSE. Moreover, induction of CD63+CD81+ and TF+ EVs in bronchial epithelial cells appears to be a universal response to various respiratory stressors. TF+ EVs may serve as biomarkers of exposure and/or risk in response to respiratory exposures and may help to guide preventive treatment decisions.

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