The blood endothelial cell chamber – An innovative system to study immune responses in drug development

Preeclampsia is characterised by an exaggerated inflammatory response and maternal endothelial cell activation. Syncytial knots, dead multinucleated fetal cells shed from the placenta in large numbers during all pregnancies, may be phagocytosed by maternal endothelial cells. Our previous studies showed that phagocytosis of necrotic but not apoptotic syncytial knots led to endothelial cell activation. It is known that phagocytosis of apoptotic cells leads to active tolerance of immune responses and in this study we questioned whether phagocytosis of apoptotic syncytial knots leads to suppression of the endothelial cells ability to be activated. Syncytial knots were harvested from 1st trimester placental explants. Monolayers of endothelial cells were pre-treated with apoptotic syncytial knots for 24 h. After washing, the endothelial cells were treated with the endothelial cell activators LPS, PMA, IL-6, or necrotic syncytial knots for 24 h. In some experiments the inhibitor of phagocytosis, cytochalasin D, was added into the cultures along with apoptotic syncytial knots. Endothelial cell-surface ICAM-1 was measured using cell based ELISAs. Expression of ICAM-1 by endothelial cells that had phagocytosed apoptotic syncytial knots prior to treatment with LPS, PMA, IL-6, or necrotic syncytial knots was significantly (P =/<0.003) reduced, compared to control endothelial cells that had not phagocytosed apoptotic syncytial knots. Inhibiting phagocytosis of apoptotic syncytial knots with cytochalasin D abolished this protective effect. Our data suggest phagocytosis of apoptotic syncytial knots results in the suppression of the ability of endothelial cells to be activated by a number of potent chemical activators, as well as by the physiologically relevant activator, necrotic syncytial knots. This work suggests that the release of apoptotic syncytial knots from the placenta during normal pregnancy may be a mechanism by which the fetus attempts to protect the maternal vasculature against activation.

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Cyclic thrombospondin-1 mimetics: grafting of a thrombospondin sequence into circular disulfide-rich frameworks to inhibit endothelial cell migration

Bioscience Reports ◽

10.1042/bsr20150210 ◽

2015 ◽

Vol 35 (6) ◽

Cited By ~ 24

Author(s):

Lai Yue Chan ◽

David J. Craik ◽

Norelle L. Daly

Keyword(s):

Cell Migration ◽

Endothelial Cell ◽

Drug Development ◽

Endothelial Cell Migration ◽

Cancer Drug ◽

Alternative Approach ◽

Thrombospondin 1 ◽

Native Fold

The findings suggest re-engineered cyclic TSP-1 mimetics are non-toxic, highly stable, and possess potent anti-angiogenesis activity without altering the native fold of the cyclic frameworks. This provides an alternative approach for cancer drug development particularly in the thrombospondin field.

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Cost-effectiveness of Adding an Agent That Improves Immune Responses to Initial Antiretroviral Therapy (ART) in HIV-Infected Patients: Guidance for Drug Development

HIV Clinical Trials ◽

10.1310/hct1301-1 ◽

2012 ◽

Vol 13 (1) ◽

pp. 1-10 ◽

Cited By ~ 4

Author(s):

Bethany L. Morris ◽

Callie A. Scott ◽

Timothy J. Wilkin ◽

Paul E. Sax ◽

Roy M. Gulick ◽

...

Keyword(s):

Cost Effectiveness ◽

Antiretroviral Therapy ◽

Immune Responses ◽

Drug Development

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A summary of meeting proceedings for ‘Measuring immune responses in non-human primates for drug development—Opportunities and challenges for predicting human efficacy and immunotoxicity’

Journal of Immunotoxicology ◽

10.3109/1547691x.2011.631610 ◽

2012 ◽

Vol 9 (1) ◽

pp. 108-120 ◽

Cited By ~ 8

Author(s):

Cris Kamperschroer ◽

Amitinder Kaur ◽

Hervé Lebrec

Keyword(s):

Immune Responses ◽

Drug Development

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Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses

Scientific Reports ◽

10.1038/s41598-021-92543-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ping Li ◽

Julia R. Walsh ◽

Kevin Lopez ◽

Abdulkadir Isidan ◽

Wenjun Zhang ◽

...

Keyword(s):

Endothelial Cell ◽

Immune Responses ◽

Cell Activation ◽

Killer Cell ◽

Genetically Engineered ◽

Organ Shortage ◽

Human Natural Killer Cell ◽

Genetic Modifications ◽

Porcine Endothelial Cell

AbstractXenotransplantation (cross-species transplantation) using genetically-engineered pig organs offers a potential solution to address persistent organ shortage. Current evaluation of porcine genetic modifications is to monitor the nonhuman primate immune response and survival after pig organ xenotransplantation. This measure is an essential step before clinical xenotransplantation trials, but it is time-consuming, costly, and inefficient with many variables. We developed an efficient approach to quickly examine human-to-pig xeno-immune responses in vitro. A porcine endothelial cell was characterized and immortalized for genetic modification. Five genes including GGTA1, CMAH, β4galNT2, SLA-I α chain, and β2-microglobulin that are responsible for the production of major xenoantigens (αGal, Neu5Gc, Sda, and SLA-I) were sequentially disrupted in immortalized porcine endothelial cells using CRISPR/Cas9 technology. The elimination of αGal, Neu5Gc, Sda, and SLA-I dramatically reduced the antigenicity of the porcine cells, though the cells still retained their ability to provoke human natural killer cell activation. In summary, evaluation of human immune responses to genetically modified porcine cells in vitro provides an efficient method to identify ideal combinations of genetic modifications for improving pig-to-human compatibility, which should accelerate the application of xenotransplantation to humans.

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Biomarkers of Endothelial Cell Activation Serve as Potential Surrogate Markers for Drug-induced Vascular Injury

Toxicologic Pathology ◽

10.1177/0192623310378866 ◽

2010 ◽

Vol 38 (6) ◽

pp. 856-871 ◽

Cited By ~ 60

Author(s):

Jun Zhang ◽

Albert F. DeFelice ◽

Joseph P. Hanig ◽

Thomas Colatsky

Keyword(s):

Endothelial Cell ◽

Drug Development ◽

Vascular Injury ◽

Cell Activation ◽

Historical Background ◽

Inflammatory Factors ◽

Surrogate Markers ◽

Endothelial Cell Activation ◽

Drug Induced ◽

Potential Biomarker

Drug-induced vascular injury (DIVI) is a nonclinical finding that often confounds the toxicological evaluation of investigational drugs, but there is an absence of qualified biomarkers that can be used to detect and monitor its appearance in animals and patients during drug development and clinical use. It is well known that endothelial cell (EC) activation plays a key role in the expression and evolution of DIVI, and the various immunological and inflammatory factors involved in its expression may serve as potential biomarker candidates. Activated ECs change their morphology and gene expression, generating endothelial adhesion molecules, pro-coagulant molecules, cytokines, chemokines, vasodilators, nitric oxide, and acute-phase reactants. This review provides a brief historical background of EC activation and the search for biomarkers of early EC activation for monitoring DIVI. At present, no biomarkers of EC activation have been qualified to predict DIVI in the nonclinical or clinical context, and a robust pathologic foundation for their use is still lacking. We propose three categories of EC activation biomarkers: recommended surrogate markers, potentially useful markers, and emerging candidate markers. This review alerts pharmaceutical companies, research institutions, and regulatory agencies to the continuing need for reliable biomarkers of EC activation in drug development.

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Endothelial cell, myeloid, and adaptive immune responses in SARS‐CoV‐2 infection

The FASEB Journal ◽

10.1096/fj.202100024r ◽

2021 ◽

Vol 35 (5) ◽

Author(s):

Nicolas Degauque ◽

Alain Haziot ◽

Sophie Brouard ◽

Nuala Mooney

Keyword(s):

Endothelial Cell ◽

Immune Responses ◽

Adaptive Immune Responses ◽

Adaptive Immune

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Publisher Correction: Genetic engineering of porcine endothelial cell lines for evaluation of human-to-pig xenoreactive immune responses

Scientific Reports ◽

10.1038/s41598-021-96406-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Ping Li ◽

Julia R. Walsh ◽

Kevin Lopez ◽

Abdulkadir Isidan ◽

Wenjun Zhang ◽

...

Keyword(s):

Endothelial Cell ◽

Genetic Engineering ◽

Immune Responses ◽

Cell Lines ◽

Porcine Endothelial Cell

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An Ultrastructural Study of Pericytes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010010007x ◽

1968 ◽

Vol 26 ◽

pp. 66-67

Author(s):

T. M. Murad ◽

E. von Haam

Keyword(s):

Plasma Membrane ◽

Endothelial Cell ◽

Basement Membrane ◽

Blood Vessel ◽

Vascular Endothelial Cells ◽

Myoepithelial Cells ◽

Capillary Blood ◽

The Body ◽

Capillary Blood Vessel ◽

Outer Plasma Membrane

Pericytes are vascular satellites present around capillary blood vessels and small venules. They have been observed in almost every tissue of the body and are thought to be related to vascular smooth muscle cells. Morphologically pericytes have great similarity to vascular endothelial cells and also slightly resemble myoepithelial cells.The present study describes the ultrastructural morphology of pericytes in normal breast tissue and in benign tumor of the breast. The study showed that pericytes are ovoid or elongated cells separated from the endothelial cell of the capillary blood vessel by the basement membrane of endothelial cell. The nuclei of pericytes are often very distinctive. Although some are round, oval, or elongated, others show marked irregularity and infolding of the nuclear membrane. The cytoplasm shows mono-or bipolar extension in which the cytoplasmic organelles are located (Fig. 1). These cytoplasmic extensions embrace the capillary blood vessel incompletely. The plasma membrane exhibits multiple areas of focal condensation called hemidesmosomes (Fig. 2, arrow). A variable number of pinocytotic vesicles are frequently seen lining the outer plasma membrane. Normally pericytes are surrounded by a basement membrane which is found more consistently on the outer plasma membrane separating the pericytes from the stromal connective tissue.

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