Effect of Alloantibody and Complement on Endothelial Cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE50112 | GEO | 2013/08/28
SECONDARY ACCESSION(S): PRJNA217335
REPOSITORIES: GEO
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Project description:Examine the possible pro-inflammatory gene effects of alloantibody and complement on endothelial cells Microarray 3 Experiment performed three times on three separate microarrays using three separate HUVEC and PRA donors. In each experiment, there are four groups of HUVEC with three replicates per group treated for 4 hours in gelatin veronal buffer containing either complement-inactivated PRA serum, vehicle (gelatin veronal buffer), PRA serum, or non-PRA serum
2013-08-28 | E-GEOD-50112 | biostudies-arrayexpress
Project description:Age-related macular degeneration (AMD) is a common, blinding disease of the elderly in which macular photoreceptor cells, retinal pigment epithelium, and choriocapillaris endothelial cells ultimately degenerate. Recent studies have found that degeneration of the choriocapillaris occurs early in this disease and that this endothelial cell dropout is concomitant with increased deposition of the complement membrane attack complex (MAC) at the choroidal endothelium. However, the impact of MAC injury to choroidal endothelial cells is poorly understood. To model this event in vitro, and to study the downstream consequences of MAC injury, endothelial cells were exposed to complement from human serum, compared to heat inactivated serum which lacks complement components. Cells exposed to complement components in human serum showed increased labeling with antibodies directed against the MAC, time and dose dependent cell death as assessed by lactate dehydrogenase assay, and increased permeability. RNA-Seq analysis following complement injury revealed increased expression of genes associated with angiogenesis including matrix metalloproteases (MMPs) 3 and 9, and VEGF-A. The MAC-induced increase in MMP9 RNA expression was validated using C5 depleted serum compared to C5 reconstitited serum. Increased levels of MMP9 were also determined using Western blot and zymography. These data suggest that, in addition to cell lysis, complement attack on choroidal endothelial cells promotes an angiogenic phenotype in surviving cells. RNA-Seq of RF/6A (cultured choroidal endothelial cells from Rhesus macaque) treated with either 50% heat-inactivated human serum ([CONTROL], n=3) or 50% normal human serum (active complement membrane attack complex [MAC], n=3)
2015-11-04 | E-GEOD-70806 | biostudies-arrayexpress
Project description:Age-related macular degeneration (AMD) is a common, blinding disease of the elderly in which macular photoreceptor cells, retinal pigment epithelium, and choriocapillaris endothelial cells ultimately degenerate. Recent studies have found that degeneration of the choriocapillaris occurs early in this disease and that this endothelial cell dropout is concomitant with increased deposition of the complement membrane attack complex (MAC) at the choroidal endothelium. However, the impact of MAC injury to choroidal endothelial cells is poorly understood. To model this event in vitro, and to study the downstream consequences of MAC injury, endothelial cells were exposed to complement from human serum, compared to heat inactivated serum which lacks complement components. Cells exposed to complement components in human serum showed increased labeling with antibodies directed against the MAC, time and dose dependent cell death as assessed by lactate dehydrogenase assay, and increased permeability. RNA-Seq analysis following complement injury revealed increased expression of genes associated with angiogenesis including matrix metalloproteases (MMPs) 3 and 9, and VEGF-A. The MAC-induced increase in MMP9 RNA expression was validated using C5 depleted serum compared to C5 reconstitited serum. Increased levels of MMP9 were also determined using Western blot and zymography. These data suggest that, in addition to cell lysis, complement attack on choroidal endothelial cells promotes an angiogenic phenotype in surviving cells.
2015-11-04 | GSE70806 | GEO
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Project description:Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare, genetic premature aging disorder associated with severe atherosclerosis, often resulting in fatal heart attacks and strokes. Progerin, the mutant protein in HGPS, also is expressed in healthy individuals and may play a role in the development of atherosclerosis during physiologic aging. Here, we provide evidence for a primary involvement of vascular endothelium in the pathogenesis of accelerated atherosclerosis in HGPS. Expression of progerin in cultured human endothelial cells induces a dysfunctional phenotype, manifested by activation of multiple pro-inflammatory, pro-atherogenic genes. In particular, our data implicate endothelial-derived interleukin-1 (IL-1) as a key mediator of a pro-inflammatory vascular phenotype. Endothelial activation also is detectable in a mouse model of HGPS, and appears to be conveyed to neighboring vascular cells via autocrine and paracrine signaling. These new mechanistic insights into the vascular pathobiology of HGPS may have therapeutic implications for this disease. Genome-wide transcriptional profiling was carried out to assess functional phenotypic changes in endothelial cells (EC) as a result of progerin expression. Cultured EC were infected with an adenovirus expressing progerin (Ad-Progerin), and as a control, an adenovirus that did not express any construct (Ad-Null). Experiments were preformed with three different EC cultures.
2010-12-02 | E-GEOD-19270 | biostudies-arrayexpress
Project description:BACKGROUND: endothelial cells (ECs) form a continuous barrier between blood and tissues and gate the traffic of molecules and cells across the vessel wall playing an active role in hemostasis, inflammation and immunity. Decidual endothelial cells (DECs) are located in a very special district from the immunological and inflammatory point of view since decidua has immunoregulatory and pro-angiogenic functions. The aim of this study was to compare the phenotype of microvascular ECs isolated from decidua with microvascular ECs isolated from other vascular districts. METHOD: we analyzed the differences in molecules involved in angiogenesis and leukocyte recruitment between DECs and endothelial cells from human skin (ADMECs) which are normally involved in inflammatory response by microarray analysis. RESULTS: our results showed that DECs are able to produce high amount of the growth factors HGF, VEGF-A and IGFBP3, important for the control of angiogenesis and express the adhesion molecules ICAM2 and ICAM3 in basal condition, important molecules for the control of leukocyte recruitment and function. We demonstrated also that DECs do not up-regulate the expression of ICAM1 in response to pro-inflammatory cytokines and produce high amount of the chemokines CXCL9/MIG and CXCL10/IP-10 in response to IFN-a. CONCLUSION: we conclude that the phenotype of these cells strongly differs from that of other endothelial cells such as ADMECs which have to be quickly able to response to infections and other pro-inflammatory stimuli. These data suggest that endothelial cells may play an active in the control of local immune response at foeto-maternal interface. Single color gene expression in human endothelial cells. Six independent experiments (3 DEC vs 3 ADMEC) were performed using different donors for each experiment.
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Project description:Shear stress is known to regulate endothelial cell orientation along the direction of flow. We asked wither cellular patterning along, in the absence of shear could have similar biological effects as shear. We used DNA microarrays to examine the effect of cellular patterning on their transcriptome.
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Project description:Neuroinflammatory and neuroimmune mechanisms, as exemplified by infiltrating immune cells and activation of resident endothelial/glial cells, respectively, are known to be involved in the establishment and maintenance of chronic pain. An immune system pathway that may be involved in the activation of both immune and glial cells is complement. The complement pathway is made up of a large number of distinct plasma proteins which react with one another to opsonize pathogens and induce a series of inflammatory responses to help fight infection. Cleaved products and complexes produced by complement activation are responsible for a range of effects including mediation of immune infiltration, activation of phagocytes, opsonization/lysis of pathogens and injured cells, and production of vasoactive amines such as histamine and serotonin. Gene-expression microarray-analysis was performed on the rat spinal nerve ligation (SNL) model of neuropathic pain Keywords: organ type comparison, disease state analysis and drug treatment
2008-03-31 | GSE10238 | GEO
Project description:Venous thromboembolism (VTE) is a common and deadly disease with unclear pathophysiological initiation. Hypoxia and inflammation are believed to contribute to promoting thrombosis, but how and if these potential triggers interact is unclear. We have stimulated primary endothelial cells with sustained and intermittent hypoxia, in the presence or absence of TNF. Effects to the cells were analyzed with RNA sequencing and flow cytometry after 24H and 48H stimulations. RNA sequencing confirmed strong pro-inflammatory effects of TNF, and indicated that hypoxia alone had modest effects, but when combined with TNF there were clear additive effects on the number of differentially expressed genes, as well as modest increases of pro-inflammatory GO terms assessed by GSEA. On protein level, flow cytometry confirmed additive effects of combingin hypoxia and TNF for several adhesion molecules including E-selectin and ICAM-1, as well as on associated molecules including tissue factor. Unsupervised clustering of the flow cytometric data also revealed the expansion of a subset of endothelial cells with strongly pro-thrombotic phenotype. Taken together, our data suggests that hypoxia combines with inflammatory cues in the in vivo situation to promote pro-thrombotic responses by endothelial cells in the venous valve pocket.
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Project description:Complement promotes vascular inflammation in transplant organ rejection and connective tissue diseases. Here we identify ZFYVE21 as a complement-induced Rab5 effector that regulates pro-inflammatory non-canonical NF-B signaling in endothelial cells (EC). In response to membrane attack complexes (MAC), ZFYVE21 is post-translationally stabilized via recruitment to MAC+Rab5+ endosomes in a Rab5- and PI(3)P-dependent manner. ZFYVE21 promotes SMURF2-mediated polyubiquitinylation and proteasome-dependent degradation of endosome-associated PTEN to induce vesicular enrichment of PI(3,4,5)P3 and sequential recruitment of activated Akt and NF-B-inducing kinase (NIK). Pharmacologic alteration of the phosphoinositide content of MAC+Rab5+ endosomes with miltefosine reduces ZFYVE21 induction, EC activation, as well as allograft vasculopathy in a humanized mouse model. ZFYVE21 induction distinctly occurs in response to MAC- but not ligand-induced non-canonical NF-B and is a biomarker for complement-mediated endothelial signaling in human renal and synovial tissues. Our data identifies ZFYVE21 as a novel Rab5 effector, defines a Rab5-ZFYVE21-SMURF2-pAkt axis by which it mediates EC activation, and demonstrates a role for this pathway as a drug target and biomarker for complement-mediated pathologies.
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