A Combined impedance and alphaLISA-based approach to identify anti-inflammatory and barrier-protective compounds in human endothelium

Maren Pflüger, Aleksandra Kapuscik, Rudolf Lucas, Anita Koppensteiner, Michael Katzlinger, Jouni Jokela, Andreas Eger, Nico Jacobi, Christoph Wiesner, Elisabeth Hofmann, Kamil Önder, Jiri Kopecky, Wolfgang Schütt, Harald Hundsberger

Research output: Contribution to journalArticlepeer-review

15 Scopus citations


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.

Original languageEnglish (US)
Pages (from-to)67-74
Number of pages8
JournalJournal of Biomolecular Screening
Issue number1
StatePublished - Jan 2013


  • AlphaLISA
  • ICAM-1
  • IL-8
  • chemokine
  • cyanobacteria
  • electrical cell substrate impedances sensing (ECIS)
  • high-throughput screening
  • inflammation
  • tumor necrosis factor (TNF)

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Molecular Medicine
  • Pharmacology
  • Drug Discovery


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