432. How do antiphospholipid antibodies contribute to preeclampsia?

2008 ◽  
Vol 20 (9) ◽  
pp. 112
Author(s):  
Q. Chen ◽  
C. Viall ◽  
P. R. Stone ◽  
L. W. Chamley
Keyword(s):  
Endothelial Cell ◽  
Antiphospholipid Antibodies ◽  
Cell Activation ◽  
Fold Increase ◽  
First Trimester ◽  
Endothelial Activation ◽  
Maternal Blood ◽  
Endothelial Cell Activation

Preeclampsia is characterised by elevated maternal blood pressure which is preceded by endothelial activation. The cause of this endothelial cell dysfunction is unclear but it appears to be triggered by a placental factor. One of the risk factors for developing preeclampsia is the presence of antiphospholipid antibodies (aPL) in the maternal blood but exactly how aPL predispose women to developing preeclampsia is unclear. A second feature known to be associated with preeclampsia is excessive shedding and deportation of dead trophoblasts. We have previously shown that shed trophoblasts are phagocytosed by endothelial cells and that phagocytosis of necrotic trophoblasts leads to endothelial cell activation1. In this study we examined the hypothesis that aPL alter the number or nature of trophoblasts shed from the placenta resulting in endothelial cell activation. Using our published model of trophoblast shedding 2 human first trimester placental explants were treated with monoclonal aPL, IIC5 or ID2, or control antibody CD45 for 72 h. Shed trophoblasts then were harvested and counted using a Cellometer AutoT4 automated cell counter. The activity of caspases 3&7 was analysed in all treated shed trophoblasts using a FLICA™ kit. The treated shed trophoblasts also were exposed to the endothelial cell line HMEC-1 for 24 h. The level of ICAM-1 by HMEC-1 was determined by cell-based ELISA. The number of trophoblasts shed from placental explants was increased 2 fold following aPL treatment whereas, treatment with CD45 resulted in only a 1.3 fold increase in shedding. Trophoblasts shed from aPL-treated explants contained less active caspases 3 & 7 compared with control shed trophoblasts. Moreover, phagocytosis of trophoblasts shed from aPL-treated explants induced significantly increased expression of ICAM-1 compared with controls. aPL treatment affected the number and nature of trophoblasts shed from placentae in such a way that phagocytosing endothelium become activated. These findings suggest that aPL treatment may have shifted the type of cell death that shed trophoblasts are undergoing from apoptosis to a more necrotic or aponecrotic mechanism. This type of shedding of trophoblasts in vivo might contribute to the endothelial cell activation which is a hallmark feature of preeclampsia. (1) Chen Q, Stone PR, McCowan LM et al. Phagocytosis of necrotic but not apoptotic trophoblasts induces endothelial cell activation. Hypertension. 2006;47:116–121. (2) Abumaree MH, Stone PR, Chamley LW. An in vitro model of human placental trophoblast deportation/shedding. Mol Hum Reprod. 2006;12:687–694.

208. Exposing necrotic trophoblasts to endothelial cells in vitro causes increased adhesion of monocytes

2005 ◽  
Vol 17 (9) ◽  
pp. 79
Author(s):  
Q. Chen ◽  
P. Stone ◽  
L. McCowan ◽  
L. Chamley
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Activation ◽  
Uv Light ◽  
Fold Increase ◽  
Maternal Blood ◽  
U937 Cells ◽  
Endothelial Cell Activation ◽  
Maternal Circulation

A number of studies suggest that there is a generalized endothelial cell activation and inflammatory response in preeclampsia, which may be caused by factors released from the placenta including deported trophoblasts. Trophoblasts are the placental cells that are bathed in maternal blood during pregnancy and as they become aged or damaged trophoblasts are shed from the placenta and deported into the maternal circulation. The fate of deported trophoblasts is unknown but we have found that endothelial cells can phagocytose dead trophoblasts. The aim of this study was to examine the effects of phagocytosing dead trophoblasts on endothelial cell–monocyte interactions. Methods: The trophoblast-derived cell lines Jar and Jeg-3 were induced to undergo necrotic death by freeze/thawing or apoptotic death by exposure to UV light. HMEC-1 endothelial cells were labeled with green fluorescent cell tracker stain and then exposed to necrotic or apoptotic trophoblasts for 3 or 24 h. U937 (monocyte) cells were labeled with red fluorescent stain and incubated with the HMEC-1 monolayers for 3 or 24 h. The adhesion of the U937 cells to the HMEC-1 monolayers was quantified by flow cytometry and compared to the adhesion of U937 cells to untreated HMEC-1 monolayers. Results: Exposing the HMEC-1 cells to necrotic, but not apoptotic, trophoblasts induced an approximately two-fold increase in the adhesion of U937 cells to the HMEC-1 monolayers (P = 0.01). The findings were consistent regardless of whether the HEMC-1 cells were exposed to the dead trophoblasts for 3 or 24 h. Conclusions: We have previously shown that endothelial cells phagocytose both apoptotic and necrotic trophoblasts. The results of the current study suggest that shedding necrotic trophoblasts from the placenta could induce endothelial cells to become activated resulting in increased leucocyte adhesion. Thus, dead trophoblasts may be one of the factors released from the placenta that induce preeclampsia.

scholarly journals Dengue Virus Induces the Expression and Release of Endocan from Endothelial Cells by an NS1–TLR4-Dependent Mechanism

2021 ◽  
Vol 9 (6) ◽  
pp. 1305
Author(s):  
Carlos Alonso Domínguez-Alemán ◽  
Luis Alberto Sánchez-Vargas ◽  
Karina Guadalupe Hernández-Flores ◽  
Andrea Isabel Torres-Zugaide ◽  
Arturo Reyes-Sandoval ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Mrna Expression ◽  
Cell Lines ◽  
Cell Activation ◽  
Dengue Infection ◽  
Elevated Serum ◽  
Fold Increase ◽  
Endothelial Cell Activation ◽  
Stimulation Of

A common hallmark of dengue infections is the dysfunction of the vascular endothelium induced by different biological mechanisms. In this paper, we studied the role of recombinant NS1 proteins representing the four dengue serotypes, and their role in promoting the expression and release of endocan, which is a highly specific biomarker of endothelial cell activation. We evaluated mRNA expression and the levels of endocan protein in vitro following the stimulation of HUVEC and HMEC-1 cell lines with recombinant NS1 proteins. NS1 proteins increase endocan mRNA expression 48 h post-activation in both endothelial cell lines. Endocan mRNA expression levels were higher in HUVEC and HMEC-1 cells stimulated with NS1 proteins than in non-stimulated cells (p < 0.05). A two-fold to three-fold increase in endocan protein release was observed after the stimulation of HUVECs or HMEC-1 cells with NS1 proteins compared with that in non-stimulated cells (p < 0.05). The blockade of Toll-like receptor 4 (TLR-4) signaling on HMEC-1 cells with an antagonistic antibody prevented NS1-dependent endocan production. Dengue-infected patients showed elevated serum endocan levels (≥30 ng/mL) during early dengue infection. High endocan serum levels were associated with laboratory abnormalities, such as lymphopenia and thrombocytopenia, and are associated with the presence of NS1 in the serum.

Biocompatibility and Inflammation Profile of B2A-Coated Granules Used in Arthrodesis

2013 ◽  
Vol 32 (2) ◽  
pp. 154-161 ◽  
Author(s):  
Paul O. Zamora ◽  
Yi Liu ◽  
Henry Guo ◽  
Xinhua Lin
Keyword(s):  
Endothelial Cell ◽  
Cell Activation ◽  
Skin Irritation ◽  
Leukocyte Recruitment ◽  
Delayed Type Hypersensitivity ◽  
Endothelial Cell Activation ◽  
Air Pouch ◽  
Coated Granules

The biocompatibility/inflammation profile of B2A-coated ceramic granules was evaluated using a panel of standard biocompatibility protocols (International Organization for Standardization-10993) including skin irritation and delayed-type hypersensitivity (Kligman maximization test), as well as acute, subacute, and chronic toxicity. Additionally, the potential of B2A-coated granules to elicit inflammatory reactions was also assessed using in vivo air pouch models, and B2A was evaluated using in vitro models of leukocyte recruitment and endothelial cell activation. Overall, the findings demonstrate that B2A-coated ceramic granules exhibit good biocompatibility profiles in the murine air pouch model and in standard subcutaneous implant models, and B2A did not demonstrate evidence of leukocyte recruitment or endothelial cell activation. These findings suggest that B2A and B2A-coated granules have little, if any, propensity to initiate inflammation reactions based on leukocyte recruitment. Thus, traditional biocompatibility and specially designed inflammation models indicate a high degree of biocompatibility and a low possibility of toxicity, inflammation, or edema following the implant of B2A-coated granules.

scholarly journals A novel flow cytometry-based quantitative monocyte adhesion assay to estimate endothelial cell activation in vitro

BioTechniques ◽  
2020 ◽  
Vol 68 (6) ◽  
pp. 325-333
Author(s):  
Vinnyfred Vincent ◽  
Himani Thakkar ◽  
Anjali Verma ◽  
Atanu Sen ◽  
Nikhil Chandran ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Activation ◽  
Endothelial Activation ◽  
Adhesion Assay ◽  
Monocyte Adhesion ◽  
Endothelial Cell Activation ◽  
Functional Level

One of the earliest events in the development of atherosclerosis is endothelial activation, which is estimated in vitro at the functional level by quantifying monocyte adhesion. This involves the incubation of fluorescently labeled monocytes on top of cultured endothelial cells and quantifying the number of adhered monocytes. Currently, the quantification of adhered monocytes is done using microscopy or by lysing the cells and estimating the fluorescence. Here we present a novel flow cytometry-based method for the quantification of monocyte adhesion. This method could quantify the average number of monocytes adhered to a single endothelial cell after monocyte adhesion assay, and was also sensitive to the level of activation of endothelial cells. Flow cytometry-based quantification requires less time and effort compared with microscopy-based quantification.

106. IL-6 INCREASES THE SHEDDING OF NECROTIC TROPHOBLASTS FROM PLACENTAL EXPLANTS

2009 ◽  
Vol 21 (9) ◽  
pp. 25
Author(s):  
Q. Chen ◽  
L. Chen ◽  
B. Liu ◽  
H. Zhao ◽  
P. R. Stone ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Activation ◽  
Elevated Serum ◽  
Serum Levels ◽  
Endothelial Activation ◽  
Maternal Blood ◽  
Endothelial Cell Activation ◽  
Cell Monolayers ◽  
Cell Dysfunction

Preeclampsia (PE) is characterised by elevated maternal blood pressure, preceded by endothelial cell dysfunction. Dead trophoblasts, shed from the placenta may be one of the factors that trigger PE. Women with PE frequently have elevated serum levels of inflammatory markers such as, IL-6 and TNF a but their functional significance is unclear. In this study we investigated whether these or other cytokines can alter trophoblast shedding from placental explants. Placental explants were treated with 9 different cytokines for 72 hours. Shed trophoblasts then were harvested using our published method1. The numbers of trophoblasts shed were quantified by automated cell counter. Expression of active of caspases 3&7 by the shed trophoblasts was determined using a FLICA kit. The trophoblasts shed from cytokine-treated or control explants were exposed to endothelial cell monolayers and endothelial activation determined by ELISA for cell surface ICAM-1. Treatment of explants with IL-6 caused a 50% increase (p=0.001), while TNF a and TGF b 1, caused smaller significant increases in the numbers of trophoblasts shed. Trophoblasts shed from explants treated with IL-6, TGF b 1, or TGF b 3 expressed significantly less active caspases 3&7 than controls or trophoblasts shed from explants treated with other cytokines. Exposing trophoblasts shed from IL-6- or TGF b 1-treated explants to endothelial cells caused a significant (P<0.001) increase in endothelial activation. Normally trophoblasts shed from the placenta die by an apoptosis-like process and their phagocytosis by endothelial cells is silent but a shift to shedding of necrotic trophoblasts can lead to endothelial cell activation 2. However, it remains unclear what might trigger a shift from apoptotic to necrotic trophoblast death. This study suggests that IL-6 and possibly other cytokines can alter both the number and the nature of shed trophoblasts such that the trophoblast are more necrotic and their phagocytosis by maternal endothelial cells could contribute to the pathogenesis of preeclampsia.

scholarly journals Functional interactions of T cells with endothelial cells: the role of CD40L-CD40-mediated signals.

1995 ◽  
Vol 182 (6) ◽  
pp. 1857-1864 ◽  
Author(s):  
M J Yellin ◽  
J Brett ◽  
D Baum ◽  
A Matsushima ◽  
M Szabolcs ◽  
...  
Keyword(s):  
T Cells ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Surface ◽  
Cell Activation ◽  
Endothelial Cell Activation ◽  
Cd40 Expression

CD40 is expressed on a variety of cells, including B cells, monocytes, dendritic cells, and fibroblasts. CD40 interacts with CD40L, a 30-33-kD activation-induced CD4+ T cell surface molecule. CD40L-CD40 interactions are known to play key roles in B cell activation and differentiation in vitro and in vivo. We now report that normal human endothelial cells also express CD40 in situ, and CD40L-CD40 interactions induce endothelial cell activation in vitro. Frozen sections from normal spleen, thyroid, skin, muscle, kidney, lung, or umbilical cord were studied for CD40 expression by immunohistochemistry. Endothelial cells from all tissues studied express CD40 in situ. Moreover, human umbilical vein endothelial cells (HUVEC) express CD40 in vitro, and recombinant interferon gamma induces HUVEC CD40 upregulation. CD40 expression on HUVEC is functionally significant because CD40L+ Jurkat T cells or CD40L+ 293 kidney cell transfectants, but not control cells, upregulate HUVEC CD54 (intercellular adhesion molecule-1), CD62E (E-selectin), and CD106 (vascular cell adhesion molecule-1) expression in vitro. Moreover, the kinetics of CD40L-, interleukin 1-, or tumor necrosis factor alpha-induced CD54, CD62E, and CD106 upregulation on HUVEC are similar. Finally, CD40L-CD40 interactions do not induce CD80, CD86, or major histocompatibility complex class II expression on HUVEC in vitro. These results demonstrate that CD40L-CD40 interactions induce endothelial cell activation in vitro. Moreover, they suggest a mechanism by which activated CD4+ T cells may augment inflammatory responses in vivo by upregulating the expression of endothelial cell surface adhesion molecules.

Imaging fibrin formation and platelet and endothelial cell activation in vivo

2011 ◽  
Vol 105 (05) ◽  
pp. 776-782 ◽  
Author(s):  
Bruce Furie ◽  
Lola Bellido-Martin ◽  
Vivien Chen ◽  
Reema Jasuja ◽  
Barbara Furie
Keyword(s):  
Blood Flow ◽  
Endothelial Cell ◽  
Animal Models ◽  
Cell Activation ◽  
In Vitro Assays ◽  
Endothelial Cell Activation ◽  
The Past ◽  
Injury Model

SummaryOver the past six decades research employing in vitro assays has identified enzymes, cofactors, cell receptors and associated ligands important to the haemostatic process and its regulation. These studies have greatly advanced our understanding of the molecular and cellular bases of haemostasis and thrombosis. However, in vitro assays cannot simultaneously reproduce the interactions of all of the components of the haemostatic process that occur in vivo nor do they reflect the importance of haemodynamic factors resulting from blood flow. To overcome these limitations investigators have increasingly turned to animal models of haemostasis and thrombosis. In this article we describe some advances in the visualisation of platelet and endothelial cell activation and blood coagulation in vivo and review what we have learned from our intravital microscopy experiments using primarily the laser-induced injury model for thrombosis.

scholarly journals Integrin αVβ3as a Target for Blocking HIV-1 Tat-Induced Endothelial Cell Activation In Vitro and Angiogenesis In Vivo

2005 ◽  
Vol 25 (11) ◽  
pp. 2315-2320 ◽  
Author(s):  
Chiara Urbinati ◽  
Stefania Mitola ◽  
Elena Tanghetti ◽  
Chandra Kumar ◽  
Johannes Waltenberger ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Activation ◽  
Endothelial Cell Activation ◽  
Hiv 1

scholarly journals The In Vitro and In Vivo Effects of Anti-Galactose Antibodies on Endothelial Cell Activation and Xenograft Rejection

2003 ◽  
Vol 170 (3) ◽  
pp. 1531-1539 ◽  
Author(s):  
Hui Xu ◽  
Dengping Yin ◽  
Bashoo Naziruddin ◽  
Libing Chen ◽  
Aileen Stark ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Activation ◽  
Endothelial Cell Activation ◽  
Xenograft Rejection ◽  
In Vivo Effects

scholarly journals Cholinergic stimulation blocks endothelial cell activation and leukocyte recruitment during inflammation

2005 ◽  
Vol 201 (7) ◽  
pp. 1113-1123 ◽  
Author(s):  
Rubina W. Saeed ◽  
Santosh Varma ◽  
Tina Peng-Nemeroff ◽  
Barbara Sherry ◽  
David Balakhaneh ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Activation ◽  
Leukocyte Recruitment ◽  
Dependent Manner ◽  
Cholinergic Stimulation ◽  
Endothelial Cell Activation ◽  
Cholinergic Agonists ◽  
Α7 Nachr

Endothelial cell activation plays a critical role in regulating leukocyte recruitment during inflammation and infection. Based on recent studies showing that acetylcholine and other cholinergic mediators suppress the production of proinflammatory cytokines via the α7 nicotinic acetylcholine receptor (α7 nAChR) expressed by macrophages and our observations that human microvascular endothelial cells express the α7 nAChR, we examined the effect of cholinergic stimulation on endothelial cell activation in vitro and in vivo. Using the Shwartzman reaction, we observed that nicotine (2 mg/kg) and the novel cholinergic agent CAP55 (12 mg/kg) inhibit endothelial cell adhesion molecule expression. Using endothelial cell cultures, we observed the direct inhibitory effects of acetylcholine and cholinergic agents on tumor necrosis factor (TNF)-induced endothelial cell activation. Mecamylamine, an nAChR antagonist, reversed the inhibition of endothelial cell activation by both cholinergic agonists, confirming the antiinflammatory role of the nAChR cholinergic pathway. In vitro mechanistic studies revealed that nicotine blocked TNF-induced nuclear factor–κB nuclear entry in an inhibitor κB (IκB)α- and IκBε-dependent manner. Finally, with the carrageenan air pouch model, both vagus nerve stimulation and cholinergic agonists significantly blocked leukocyte migration in vivo. These findings identify the endothelium, a key regulator of leukocyte trafficking during inflammation, as a target of anti-inflammatory cholinergic mediators.

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