Pub Date : 1996-01-01DOI: 10.3109/10623329609024696
L. Thomsen, D. Daugaard, Helle K. Iversen, J. Olesen
Increasing evidence suggests that nitric oxide (NO) plays a role in migraine pain. Thus, the NO donor glyceryl trinitrate provokes migraine attacks in sufferers and a greater dilatation of large arteries than in controls. Physiological NO-mediated dilatation of large arteries might therefore be abnormal in migraine. Recently an ultrasound method for the detection of shear stress stimulated endothelial NO release in large arteries in man has been described. Using this method we compared radial artery dilatation during reactive hyperaernia in 12 female sufferers of migraine without aura and 12 age- and sex- matched healthy subjects. No group differences were detected in these responses (peak dilatation 111 ± 2% (mean ± SEM) of baseline in migraineurs and 112 ± 3% of baseline in the healthy control group, p = 0.79). These results indicate that shear stress stimulated endothelial NO release is normal in peripheral conduit arteries in patients suffering from migraine without aura. It remains to be settled whet...
{"title":"Normal Radial Artery Dilatation During Reactive Hyperaemia in Migraine without Aura","authors":"L. Thomsen, D. Daugaard, Helle K. Iversen, J. Olesen","doi":"10.3109/10623329609024696","DOIUrl":"https://doi.org/10.3109/10623329609024696","url":null,"abstract":"Increasing evidence suggests that nitric oxide (NO) plays a role in migraine pain. Thus, the NO donor glyceryl trinitrate provokes migraine attacks in sufferers and a greater dilatation of large arteries than in controls. Physiological NO-mediated dilatation of large arteries might therefore be abnormal in migraine. Recently an ultrasound method for the detection of shear stress stimulated endothelial NO release in large arteries in man has been described. Using this method we compared radial artery dilatation during reactive hyperaernia in 12 female sufferers of migraine without aura and 12 age- and sex- matched healthy subjects. No group differences were detected in these responses (peak dilatation 111 ± 2% (mean ± SEM) of baseline in migraineurs and 112 ± 3% of baseline in the healthy control group, p = 0.79). These results indicate that shear stress stimulated endothelial NO release is normal in peripheral conduit arteries in patients suffering from migraine without aura. It remains to be settled whet...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"1 1","pages":"199-206"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88704100","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1996-01-01DOI: 10.3109/10623329609024681
D. Lang, A. Shah, M. Lewis
Vascular endothelial cells release bradykinin (BK), which acts in an autocrine manner to raise intracellular calcium concentrations ([Ca2+]i) and release NO, effects which can be modified by angiotensin-converting enzyme (ACE) inhibitors. Little is known of the effects of BK and ACE inhibitors on endocardial endothelium however. In this study, we investigated the effects of the ACE inhibitors captopril and ramiprilat on (a) BK-degrading activity, (b) resting Ca2+ homeostasis and (c) BK-induced changes in [Ca2+]i and cGMP in bovine aortic endothelial (BAE) and right ventricle endocardial (BRVE) cells. Captopril and ramiprilat inhibited the BK-degrading activity of both cell types. Resting levels of [Ca2+]i and cGMP were significantly higher in BRVE than in BAE. Captopril or ramiprilat did not alter resting [Ca*+]i, but caused significant increases in resting cGMP which were reduced by the NO synthase inhibitor L-nitroarginine benzyl ester and the B2-kinin receptor antagonist HOE 140. Captopril and ramipril...
{"title":"Aniotensin-Converting Enzyme (ACE) Activity: Aortic ancf Endocardial Endothelium Compared","authors":"D. Lang, A. Shah, M. Lewis","doi":"10.3109/10623329609024681","DOIUrl":"https://doi.org/10.3109/10623329609024681","url":null,"abstract":"Vascular endothelial cells release bradykinin (BK), which acts in an autocrine manner to raise intracellular calcium concentrations ([Ca2+]i) and release NO, effects which can be modified by angiotensin-converting enzyme (ACE) inhibitors. Little is known of the effects of BK and ACE inhibitors on endocardial endothelium however. In this study, we investigated the effects of the ACE inhibitors captopril and ramiprilat on (a) BK-degrading activity, (b) resting Ca2+ homeostasis and (c) BK-induced changes in [Ca2+]i and cGMP in bovine aortic endothelial (BAE) and right ventricle endocardial (BRVE) cells. Captopril and ramiprilat inhibited the BK-degrading activity of both cell types. Resting levels of [Ca2+]i and cGMP were significantly higher in BRVE than in BAE. Captopril or ramiprilat did not alter resting [Ca*+]i, but caused significant increases in resting cGMP which were reduced by the NO synthase inhibitor L-nitroarginine benzyl ester and the B2-kinin receptor antagonist HOE 140. Captopril and ramipril...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"18 1","pages":"51-61"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86598528","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1996-01-01DOI: 10.3109/10623329609024682
N. Hill, B. Pierchala, A. Johns, F. Kiechle, G. Rubanyi, T. Malinski
A porphyrinic sensor was developed and used to monitor the release of nitric oxide (NO) by a fetal bovine aorta endothelial (FBAE) cell culture and a primary bovine aorta endothelial (BAE) cell culture, with populations of several thousand cells. The sensor consisted of either a platinum mesh or reticulated vitreous carbon support covered with several layers of a p-type semiconducting metalloporphyrin and a cation exchanger, Nafion. Tissue and cell cultures can be grown directly on the surface of the sensor and NO release can be measured by amperometric or differential pulse voltammetry methods. The detection limit of the sensor deposited on reticulated vitrous carbon and platinum mesh support is 10 nM and 0.1 μM respectively. The response time is about one millisecond and precision 5-6%. The peak NO concentration released from FBAE cells was 190 ± 20 nM and 70 ± 20 nM when agonized by calcium ionophore (A23187) and bradykinin respectively. The maximum NO concentration released from the primary culture BA...
{"title":"In Situ Measurements of Nitric Oxide Release from Endothelial Cells Grown Directly on a Porphyrinic Sensor","authors":"N. Hill, B. Pierchala, A. Johns, F. Kiechle, G. Rubanyi, T. Malinski","doi":"10.3109/10623329609024682","DOIUrl":"https://doi.org/10.3109/10623329609024682","url":null,"abstract":"A porphyrinic sensor was developed and used to monitor the release of nitric oxide (NO) by a fetal bovine aorta endothelial (FBAE) cell culture and a primary bovine aorta endothelial (BAE) cell culture, with populations of several thousand cells. The sensor consisted of either a platinum mesh or reticulated vitreous carbon support covered with several layers of a p-type semiconducting metalloporphyrin and a cation exchanger, Nafion. Tissue and cell cultures can be grown directly on the surface of the sensor and NO release can be measured by amperometric or differential pulse voltammetry methods. The detection limit of the sensor deposited on reticulated vitrous carbon and platinum mesh support is 10 nM and 0.1 μM respectively. The response time is about one millisecond and precision 5-6%. The peak NO concentration released from FBAE cells was 190 ± 20 nM and 70 ± 20 nM when agonized by calcium ionophore (A23187) and bradykinin respectively. The maximum NO concentration released from the primary culture BA...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"22 1","pages":"63-69"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76018787","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1996-01-01DOI: 10.3109/10623329609024701
A. Koller, G. Kaley
For a long time, locally released metabolic factors from parenchymal cells and the myo-genic response of vascular smooth muscle were considered to be the two main peripheral regulatory mechanisms to control vascular resistance and thereby determining the distribution of blood flow and pressure within vascular networks of various organs of the body. However, not every change in blood flow could be satisfactorily explained by these two mechanisms. The “flow-dependent” responses of blood vessels, namely, an increase in blood flow in arteries followed by their dilation had already been observed at the beginning of this century, and perhaps even earlier, yet the nature and importance of this phenomenon in blood flow regulation was not delineated until recently. In the last two decades, especially following the recognition of the role of endothelium in the production of vasoactive factors, much new experimental evidence was gathered that suggests the general presence and importance of flow (shear stress)-depend...
{"title":"Shear Stress Dependent Regulation of Vascular Resistance in Health and Disease: Role of Endothelium","authors":"A. Koller, G. Kaley","doi":"10.3109/10623329609024701","DOIUrl":"https://doi.org/10.3109/10623329609024701","url":null,"abstract":"For a long time, locally released metabolic factors from parenchymal cells and the myo-genic response of vascular smooth muscle were considered to be the two main peripheral regulatory mechanisms to control vascular resistance and thereby determining the distribution of blood flow and pressure within vascular networks of various organs of the body. However, not every change in blood flow could be satisfactorily explained by these two mechanisms. The “flow-dependent” responses of blood vessels, namely, an increase in blood flow in arteries followed by their dilation had already been observed at the beginning of this century, and perhaps even earlier, yet the nature and importance of this phenomenon in blood flow regulation was not delineated until recently. In the last two decades, especially following the recognition of the role of endothelium in the production of vasoactive factors, much new experimental evidence was gathered that suggests the general presence and importance of flow (shear stress)-depend...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"12 1","pages":"247-272"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81313762","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1996-01-01DOI: 10.3109/10623329609024687
Alan W. Stitt, T. Gardiner, J. Bailie, U. Chakravarthy, D. Archer
Confluent monolayers of cultured retinal microvascular endothelial cells (RVEC's) were exposed simultaneously to two different protein-gold conjugates to determine if partitioning or sorting of protein ligands occurs during receptor mediated endocytosis in CNS microvascular cells. A mixture of bovine serum albumin (BSA), conjugated with 5nm colloidal gold (BSA-gold) and transferrin (TO, conjugated with 15nm gold (Tf-gold) were added to RVEC's and the monolayers processed, in situ, for TEM. At 0 mins the gold conjugates bound diffusely to the apical plasma membrane, but after 2 mins, both ligands were observed to cluster in clathrin-coated pits prior to internalisation in coated vesicles. At 5 mins the gold-ligands were co-localised in coated vesicles and early endosomes. After 10 mins the probes were observed in larger late endosomes where segregation was first noted and by 30mins, the mixture of gold particles appeared partitioned within late endosomes, with BSA-gold localised at the periphery and Tf-gol...
{"title":"An Investigation of Receptor-Mediated Endocytosis and Endosomal Sorting of Albumin and Transferrin in Retinal Vascular Endothelial Cells","authors":"Alan W. Stitt, T. Gardiner, J. Bailie, U. Chakravarthy, D. Archer","doi":"10.3109/10623329609024687","DOIUrl":"https://doi.org/10.3109/10623329609024687","url":null,"abstract":"Confluent monolayers of cultured retinal microvascular endothelial cells (RVEC's) were exposed simultaneously to two different protein-gold conjugates to determine if partitioning or sorting of protein ligands occurs during receptor mediated endocytosis in CNS microvascular cells. A mixture of bovine serum albumin (BSA), conjugated with 5nm colloidal gold (BSA-gold) and transferrin (TO, conjugated with 15nm gold (Tf-gold) were added to RVEC's and the monolayers processed, in situ, for TEM. At 0 mins the gold conjugates bound diffusely to the apical plasma membrane, but after 2 mins, both ligands were observed to cluster in clathrin-coated pits prior to internalisation in coated vesicles. At 5 mins the gold-ligands were co-localised in coated vesicles and early endosomes. After 10 mins the probes were observed in larger late endosomes where segregation was first noted and by 30mins, the mixture of gold particles appeared partitioned within late endosomes, with BSA-gold localised at the periphery and Tf-gol...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"10 1","pages":"113-118"},"PeriodicalIF":0.0,"publicationDate":"1996-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87777987","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-05-01DOI: 10.3109/10623329509053385
S. V. Ijzendoorn, J. Heemskerk, C. Reutelingsperger
Hemostasis and thrombosis occur by cooperative actions of blood cells and cells from the vessel wall. Platelets and endothelial cells are crucial in both processes and are thought to communicate not only during the formation of a thrombus, but also while the blood is circulating. Recent investigations have suggested that the mutual interactions between these cells reach a high degree of complexity under (patho)physiological conditions. Intercellular communication can be effected in a number of ways: (i) by the constitutive release of messenger molecules from unstimulated platelets and endothelial cells, (ii) by the regulated release of messengers from stimulated cells, and (iii) by the effects of vascular tonus and blood flow on both cells. This review focusses on what is known about the signal transduction mechanisms, by which the compounds released from platelets and endothelial cells influence each others functions and properties. Special attention is also paid to the recently described effects of mech...
{"title":"Interactions between Endothelial Cells and Blood Platelets","authors":"S. V. Ijzendoorn, J. Heemskerk, C. Reutelingsperger","doi":"10.3109/10623329509053385","DOIUrl":"https://doi.org/10.3109/10623329509053385","url":null,"abstract":"Hemostasis and thrombosis occur by cooperative actions of blood cells and cells from the vessel wall. Platelets and endothelial cells are crucial in both processes and are thought to communicate not only during the formation of a thrombus, but also while the blood is circulating. Recent investigations have suggested that the mutual interactions between these cells reach a high degree of complexity under (patho)physiological conditions. Intercellular communication can be effected in a number of ways: (i) by the constitutive release of messenger molecules from unstimulated platelets and endothelial cells, (ii) by the regulated release of messengers from stimulated cells, and (iii) by the effects of vascular tonus and blood flow on both cells. This review focusses on what is known about the signal transduction mechanisms, by which the compounds released from platelets and endothelial cells influence each others functions and properties. Special attention is also paid to the recently described effects of mech...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"28 1","pages":"81-98"},"PeriodicalIF":0.0,"publicationDate":"1995-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79280321","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024658
H. Sqalli-Houssaini, F. Martin‐Nizard, E. Walters-Laporte, A. Boullier, R. Mackereel, J. Fruchart, P. Duriez
An increased level of low density lipoproreins (LDL) in the blood and an accumulation of oxidized-LDL (ox-LDL) in the subendothelium are well known factors of atherosclerosis. Endothelin (ET), a potent vasoconstrictor isolated from endothelial cells, is mitogenic for smooth muscle cells and may participate in atherogenesis by this process. In the present paper we study the effects of native LDL and ox-LDL on ET secretion by adult bovine aortic endothelial cells (ABAE). Native LDL stimulate ET secretion in a dose dependent manner between 25 and 200 μg/ml, while the effects of ox-LDL are complex. A low concentration (50 μg/ml) of highly ox-LDL induces ET secretion while a high concentration (100 μg/ml) does not induce the secretion and is cytotoxic. 100 μ/ml of slightly ox-LDL stimulates the secretion. Our data indicated that the LDL(B/E) receptor pathway is not involved in n-LDL induced ET secretion and are not conclusive on the role of the scavenger receptor pathway in ox-LDL induced ET secretion. Lysolec...
{"title":"Oxidised LDL Modulates Endothelin Secretion by Adult Bovine Aortic Endothelial Cells","authors":"H. Sqalli-Houssaini, F. Martin‐Nizard, E. Walters-Laporte, A. Boullier, R. Mackereel, J. Fruchart, P. Duriez","doi":"10.3109/10623329509024658","DOIUrl":"https://doi.org/10.3109/10623329509024658","url":null,"abstract":"An increased level of low density lipoproreins (LDL) in the blood and an accumulation of oxidized-LDL (ox-LDL) in the subendothelium are well known factors of atherosclerosis. Endothelin (ET), a potent vasoconstrictor isolated from endothelial cells, is mitogenic for smooth muscle cells and may participate in atherogenesis by this process. In the present paper we study the effects of native LDL and ox-LDL on ET secretion by adult bovine aortic endothelial cells (ABAE). Native LDL stimulate ET secretion in a dose dependent manner between 25 and 200 μg/ml, while the effects of ox-LDL are complex. A low concentration (50 μg/ml) of highly ox-LDL induces ET secretion while a high concentration (100 μg/ml) does not induce the secretion and is cytotoxic. 100 μ/ml of slightly ox-LDL stimulates the secretion. Our data indicated that the LDL(B/E) receptor pathway is not involved in n-LDL induced ET secretion and are not conclusive on the role of the scavenger receptor pathway in ox-LDL induced ET secretion. Lysolec...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"03 1","pages":"47-55"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85947964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024655
M. Ott, J. Olson, B. Ballermann
Endothelial cells in vivo are exposed to flowing blood and are therefore subject to continuous apical shear stress. This study explored the feasibility of endothelial cell culture with chronic shear stress. Bovine glomerular capillary and aortic endothelial cells were cultured under conventional conditions without flow, or in polypropylene hollow fibers perfused for 9 days with venous or arterial shear stress. Cells were then examined by scanning and transmission electron microscopy. For both cell types, ultrastructural differences between conventional culture and culture with chronic arterial flow were quantified morphometrically. In the hollow fibers, capillary and aortic endothelial cells formed adherent, confluent monolayers with chronic flow, but not without flow. With venous and arterial shear stress, aortic, but not glomerular capillary endothelial cells, aligned themselves in the direction of flow. In aortic endothelial cells, the density of Weibel Palade bodies was, on average, 38 fold greater wi...
{"title":"Chronic In Vitro Flow Promotes Ultrastructural Differentiation of Endothelial Cells","authors":"M. Ott, J. Olson, B. Ballermann","doi":"10.3109/10623329509024655","DOIUrl":"https://doi.org/10.3109/10623329509024655","url":null,"abstract":"Endothelial cells in vivo are exposed to flowing blood and are therefore subject to continuous apical shear stress. This study explored the feasibility of endothelial cell culture with chronic shear stress. Bovine glomerular capillary and aortic endothelial cells were cultured under conventional conditions without flow, or in polypropylene hollow fibers perfused for 9 days with venous or arterial shear stress. Cells were then examined by scanning and transmission electron microscopy. For both cell types, ultrastructural differences between conventional culture and culture with chronic arterial flow were quantified morphometrically. In the hollow fibers, capillary and aortic endothelial cells formed adherent, confluent monolayers with chronic flow, but not without flow. With venous and arterial shear stress, aortic, but not glomerular capillary endothelial cells, aligned themselves in the direction of flow. In aortic endothelial cells, the density of Weibel Palade bodies was, on average, 38 fold greater wi...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"10 1","pages":"21-30"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85950668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509024664
M. Watts, P. Hewett, M. Woodcock
The isolation and characterisation of human microvascular endothelial cells from adipose tissue is described and a comparison is made between isolations from abdominal subcutaneous and omental adipose tissue. Microvascular cells were isolated by incubating finely chopped material in (i) collagenase type II solution, followed by density gradient centrifu-gation and further incubation in trypsin, or in (ii) collagenase type II solution, followed by trypsin incubation and selective filtration. The final step in both procedures was selection using Ulex europaeus agglutinin-1 coated Dynabeads. The cells were characterised using immunocytochemical and biochemical techniques. The cells exhibited characteristic staining for antigens recognised by antibodies against von Willebrand factor and platelet-endothelial cell adhesion molecule (PECAM-I). They also expressed angiotensin-converting enzyme and internalised acetylated low-density lipoprotein.
{"title":"Isolation, Culture and Identification of Human Microvascular Endothelial Cells: A Comparison of Abdominal Subcutaneous and Omental Adipose Tissue","authors":"M. Watts, P. Hewett, M. Woodcock","doi":"10.3109/10623329509024664","DOIUrl":"https://doi.org/10.3109/10623329509024664","url":null,"abstract":"The isolation and characterisation of human microvascular endothelial cells from adipose tissue is described and a comparison is made between isolations from abdominal subcutaneous and omental adipose tissue. Microvascular cells were isolated by incubating finely chopped material in (i) collagenase type II solution, followed by density gradient centrifu-gation and further incubation in trypsin, or in (ii) collagenase type II solution, followed by trypsin incubation and selective filtration. The final step in both procedures was selection using Ulex europaeus agglutinin-1 coated Dynabeads. The cells were characterised using immunocytochemical and biochemical techniques. The cells exhibited characteristic staining for antigens recognised by antibodies against von Willebrand factor and platelet-endothelial cell adhesion molecule (PECAM-I). They also expressed angiotensin-converting enzyme and internalised acetylated low-density lipoprotein.","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"99 1","pages":"181-188"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87580695","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 1995-01-01DOI: 10.3109/10623329509053390
J. Langner, D. Riemann, H. Machulla
Using cultivated human umbilical vein endothelial cells (HUVEC) as a model, we investigated a selection of basic biochemical and cell physiological functions with relevance to antigen processing and presentation, namely pinocytosis and phagocytosis, the presence and activity of proteinases and the expression of MHC class II molecules. Antigen presentation (tetanus toxoid) to HLA-matched T lymphocytes with addition of proteinase inhibitors (E-64 for cysteine, pepstatin for aspartate proteinases) to HUVEC during the antigen handling phase resulted in a strongly reduced T lymphocyte stimulation by this antigen. This is taken as the first reported evidence for an antigen processing step taking place in endothelial cells. The involvement of cysteine as well as aspartate proteinases in antigen processing has been found also for other antigen presenting cells like macrophages or B lymphoblastic cells, therefore antigen processing in HUVEC most probably follows the consensus pathway formulated for these cells: up...
{"title":"Antigen Processing and Presentation by Vascular Endothelial Cells: Biochemical and Cell Physiological Preconditions","authors":"J. Langner, D. Riemann, H. Machulla","doi":"10.3109/10623329509053390","DOIUrl":"https://doi.org/10.3109/10623329509053390","url":null,"abstract":"Using cultivated human umbilical vein endothelial cells (HUVEC) as a model, we investigated a selection of basic biochemical and cell physiological functions with relevance to antigen processing and presentation, namely pinocytosis and phagocytosis, the presence and activity of proteinases and the expression of MHC class II molecules. Antigen presentation (tetanus toxoid) to HLA-matched T lymphocytes with addition of proteinase inhibitors (E-64 for cysteine, pepstatin for aspartate proteinases) to HUVEC during the antigen handling phase resulted in a strongly reduced T lymphocyte stimulation by this antigen. This is taken as the first reported evidence for an antigen processing step taking place in endothelial cells. The involvement of cysteine as well as aspartate proteinases in antigen processing has been found also for other antigen presenting cells like macrophages or B lymphoblastic cells, therefore antigen processing in HUVEC most probably follows the consensus pathway formulated for these cells: up...","PeriodicalId":11588,"journal":{"name":"Endothelium-journal of Endothelial Cell Research","volume":"23 1","pages":"141-150"},"PeriodicalIF":0.0,"publicationDate":"1995-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84627823","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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