TY - JOUR
T1 - A Combined impedance and alphaLISA-based approach to identify anti-inflammatory and barrier-protective compounds in human endothelium
AU - Pflüger, Maren
AU - Kapuscik, Aleksandra
AU - Lucas, Rudolf
AU - Koppensteiner, Anita
AU - Katzlinger, Michael
AU - Jokela, Jouni
AU - Eger, Andreas
AU - Jacobi, Nico
AU - Wiesner, Christoph
AU - Hofmann, Elisabeth
AU - Önder, Kamil
AU - Kopecky, Jiri
AU - Schütt, Wolfgang
AU - Hundsberger, Harald
N1 - Funding Information:
The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was funded by the European Territorial Co-operation grant (M00140) and the Österreichische Forschungsförderungsgesellschaft (FFG, 821021, 822710).
PY - 2013/1
Y1 - 2013/1
N2 - Chronic inflammation is at least partially mediated by the chemokine-mediated attraction and by the adhesion molecule-directed binding of leukocytes to the activated endothelium. Therefore, it is therapeutically important to identify anti-inflammatory compounds able to control the interaction between leukocytes and the endothelial compartments of the micro- and macrocirculation. When testing novel drug candidates, it is, however, of the utmost importance to detect side effects, such as potential cytotoxic and barrier-disruptive activities. Indeed, minor changes in the endothelial monolayer integrity may increase the permeability of small blood vessels and capillaries, which, in extreme cases, can lead to edema development. Here, we describe the development of a high-throughput screening (HTS) platform, based on AlphaLISA technology, able to identify anti-inflammatory nontoxic natural or synthetic compounds capable of reducing tumor necrosis factor (TNF)-induced chemokine (interleukin [IL]-8) and adhesion molecule (ICAM-1) expression in human lung microvascular endothelial cells. Quantification of cell membrane-expressed ICAM-1 and of cell culture supernatant-associated levels of IL-8 was analyzed in HTS. In parallel, we monitored monolayer integrity and endothelial cell viability using the electrical cell substrate impedance sensing method. This platform allowed us to identify natural secondary metabolites from cyanobacteria, capable of reducing ICAM-1 and IL-8 levels in TNF-activated human microvascular endothelial cells in the absence of endothelial monolayer barrier disruption.
AB - Chronic inflammation is at least partially mediated by the chemokine-mediated attraction and by the adhesion molecule-directed binding of leukocytes to the activated endothelium. Therefore, it is therapeutically important to identify anti-inflammatory compounds able to control the interaction between leukocytes and the endothelial compartments of the micro- and macrocirculation. When testing novel drug candidates, it is, however, of the utmost importance to detect side effects, such as potential cytotoxic and barrier-disruptive activities. Indeed, minor changes in the endothelial monolayer integrity may increase the permeability of small blood vessels and capillaries, which, in extreme cases, can lead to edema development. Here, we describe the development of a high-throughput screening (HTS) platform, based on AlphaLISA technology, able to identify anti-inflammatory nontoxic natural or synthetic compounds capable of reducing tumor necrosis factor (TNF)-induced chemokine (interleukin [IL]-8) and adhesion molecule (ICAM-1) expression in human lung microvascular endothelial cells. Quantification of cell membrane-expressed ICAM-1 and of cell culture supernatant-associated levels of IL-8 was analyzed in HTS. In parallel, we monitored monolayer integrity and endothelial cell viability using the electrical cell substrate impedance sensing method. This platform allowed us to identify natural secondary metabolites from cyanobacteria, capable of reducing ICAM-1 and IL-8 levels in TNF-activated human microvascular endothelial cells in the absence of endothelial monolayer barrier disruption.
KW - AlphaLISA
KW - ICAM-1
KW - IL-8
KW - chemokine
KW - cyanobacteria
KW - electrical cell substrate impedances sensing (ECIS)
KW - high-throughput screening
KW - inflammation
KW - tumor necrosis factor (TNF)
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U2 - 10.1177/1087057112458316
DO - 10.1177/1087057112458316
M3 - Article
C2 - 22941294
AN - SCOPUS:84871540580
SN - 1087-0571
VL - 18
SP - 67
EP - 74
JO - Journal of Biomolecular Screening
JF - Journal of Biomolecular Screening
IS - 1
ER -