The HIV-Tat protein interacts with Sp3 transcription factor and inhibits its binding to a distal site of the sod2 promoter in human pulmonary artery endothelial cells

Expression of cell adhesion molecule in endothelial cells upon activation by human immunodeficiency virus (HIV) infection is associated with the development of atherosclerotic vasculopathy. We postulated that induction of vascular cell adhesion molecule-1 (VCAM-1) by HIV-1 Tat protein in endothelial cells might represent an early event that could culminate in inflammatory cell recruitment and vascular injury. We determined the role of HIV-1 Tat protein in VCAM-1 expression in human pulmonary artery endothelial cells (HPAEC). HIV-1 Tat protein treatment significantly increased cell-surface expression of VCAM-1 in HPAEC. Consistently, mRNA expression of VCAM-1 was also increased by HIV-1 Tat protein as measured by RT-PCR. HIV-1 Tat protein-induced VCAM-1 expression was abolished by the NF-κB inhibitor pyrrolidine dithiocarbamate (PDTC) and the p38 MAPK inhibitor SB-203580. Furthermore, HIV-1 Tat protein enhanced DNA binding activity of NF-κB, facilitated nuclear translocation of NF-κB subunit p65, and increased production of reactive oxygen species (ROS). Similarly to VCAM-1 expression, HIV-1 Tat protein-induced NF-κB activation and ROS generation were abrogated by PDTC and SB-203580. These data indicate that HIV-1 Tat protein is able to induce VCAM-1 expression in HPAEC, which may represent a pivotal early molecular event in HIV-induced vascular/pulmonary injury. These data also suggest that the molecular mechanism underlying the HIV-1 Tat protein-induced VCAM-1 expression may involve ROS generation, p38 MAPK activation, and NF-κB translocation, which are the characteristics of pulmonary endothelial cell activation.

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72 ACTIVATED PROTEIN C ATTENUATES THROMBIN-INDUCED BARRIER DYSFUNCTION IN HUMAN PULMONARY ARTERY ENDOTHELIAL CELLS.

Journal of Investigative Medicine ◽

10.1136/jim-52-suppl2-72 ◽

2004 ◽

Vol 52 (Suppl 2) ◽

pp. S358.1-S358

Author(s):

J. H. Finigan ◽

S. M. Dudek ◽

S. Q. Ye ◽

J. G. Garcia

Keyword(s):

Endothelial Cells ◽

Pulmonary Artery ◽

Protein C ◽

Activated Protein C ◽

Barrier Dysfunction ◽

Human Pulmonary Artery

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Selective biological response of human pulmonary microvascular endothelial cells and human pulmonary artery smooth muscle cells on cold-plasma-modified polyester vascular prostheses

Biomedical Materials ◽

10.1088/1748-6041/6/6/065003 ◽

2011 ◽

Vol 6 (6) ◽

pp. 065003 ◽

Cited By ~ 8

Author(s):

N Blanchemain ◽

M R Aguilar ◽

F Chai ◽

M Jimenez ◽

E Jean-Baptiste ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Smooth Muscle Cells ◽

Cold Plasma ◽

Biological Response ◽

Microvascular Endothelial Cells ◽

Vascular Prostheses ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery

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Fenfluramine-Induced Gene Dysregulation in Human Pulmonary Artery Smooth Muscle and Endothelial Cells

Pulmonary Circulation ◽

10.4103/2045-8932.87310 ◽

2011 ◽

Vol 1 (3) ◽

pp. 405-418 ◽

Cited By ~ 5

Author(s):

Weijuan Yao ◽

Wenbo Mu ◽

Amy Zeifman ◽

Michelle Lofti ◽

Carmelle V. Remillard ◽

...

Keyword(s):

Endothelial Cells ◽

Smooth Muscle ◽

Pulmonary Artery ◽

Gene Dysregulation ◽

Pulmonary Artery Smooth Muscle ◽

Human Pulmonary Artery

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Abstract 218: Fibrinolytic Reactivity of Venous Endothelial Cells from Different Vascular Beds

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.32.suppl_1.a218 ◽

2012 ◽

Vol 32 (suppl_1) ◽

Author(s):

Neil G Kumar ◽

Elisa Roztocil ◽

John P Cullen ◽

David L Gillespie

Keyword(s):

Endothelial Cells ◽

Pulmonary Artery ◽

Cell Types ◽

Iliac Vein ◽

Vein Thrombosis ◽

Cellular Expression ◽

Artery Embolism ◽

Vascular Beds ◽

Human Pulmonary Artery ◽

Pai 1

Objective: Little is known about the molecular biology of endothelial cells from different venous vascular beds. As a result, our treatment of deep vein thrombosis (DVT) and pulmonary artery embolism (PE) remain identical. PAI-1 and tPA are important regulators of thrombosis and fibrinolysis, while ICAM-1 is known to bind fibrinogen. Here, we aim to investigate differences in fibrinolytic reactivity between human iliac vein endothelial cells (HIVECs) and human pulmonary artery endothelial cells (HPAECs). Methods: Confluent HIVECs and HPAECs, passages 3 - 6, were cultured in the absence or presence of TNFα (10 ng/mL) for 24 hours. Cellular expression of tPA and PAI-1 as analyzed by Western blot analysis and ICAM-1 as analyzed by flow cytometry were compared to controls. Results: Following TNFα stimulation, PAI-1 was upregulated in both HPAECs and HIVECs, however the upregulation observed in HPAECs was approximately 9-fold the increase observed in HIVECs (relative expression: 3.23 ± 0.52 vs 1.26 ± 0.27, n = 3, p < 0.05). While TNFα had no effect on tPA expression in HIVECs, tPA expression in HPAECS was upregulated by 33% (n = 3, p < 0.05). Although TNFα stimulation increased the number of ICAM-1 positive to approximately 100% in both cell types, a 3-fold greater increase in the Mean Fluorescence Intensity (MFI) was observed in HIVECs when compared to HPAECs (relative MFI: 69.28 ± 13.58 vs 21.92 ± 7.22, n = 3, p <0.05). Conclusions: HPAECs and HIVECs react differently in terms of fibrinolytic potential when challenged with a cytokine associated with systemic inflammation, such as in DVT and PE. These findings suggest that endothelial cells from distinct venous vascular beds may differentially regulate the fibrinolytic pathway, thus demonstrating unique properties of the deep veins and the pulmonary artery to respond to thromboembolism.

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Isolation and maintenance of human pulmonary artery endothelial cells in culture isolated from transplant donors

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1994.267.4.l406 ◽

1994 ◽

Vol 267 (4) ◽

pp. L406-L413 ◽

Cited By ~ 1

Author(s):

G. A. Visner ◽

E. D. Staples ◽

S. E. Chesrown ◽

E. R. Block ◽

D. S. Zander ◽

...

Keyword(s):

Endothelial Cells ◽

Pulmonary Artery ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Immunofluorescent Staining ◽

Northern Analysis ◽

Pulmonary Vasculature ◽

Pulmonary Vascular Disease ◽

Antigen Uptake ◽

Human Pulmonary Artery

Even though endothelial cells from different locations have similarities, there are potential morphological and functional differences between cells from different vascular regions, as well as between species. Our laboratory is interested in studying the molecular regulation of vasoactive substances in pulmonary vasculature. Therefore, we have developed reproducible methodology to isolate and maintain cultures of human pulmonary artery endothelial cells. The major innovation has been the employment of sections of pulmonary artery from heart transplant donors, from which endothelial cells are isolated. Cell monolayers were identified as endothelial cells by phase-contrast microscopy. Representative dishes of cells were further characterized by indirect immunofluorescent staining for factor VIII antigen, uptake of acetylated low-density lipoprotein, and electron microscopy. These cells were also evaluated for the expression of endothelin-1 (ET-1), a vasoactive 21-amino acid peptide derived from endothelial cells. The cells expressed ET-1 peptide and mRNA as determined by radioimmunoassay and Northern analysis, respectively. We also demonstrated that these cells are useful in transient transfection experiments for potential evaluation of promoter elements. The availability and relevance of these cells provide an important investigative tool for studies on human pulmonary vascular disease.

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