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FTY720-induced suicidal erythrocyte death

Matthias Eberhard1, Klaus Ferlinz, Katia Alizzi

  • 1Department of Physiology, University of Tübingen, Tübingen, Germany.

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology
|November 11, 2010
PubMed

Insights

FTY720, an anti-inflammatory drug, induces suicidal erythrocyte death (eryptosis) by increasing intracellular calcium. This effect, observed at toxic concentrations, involves cell membrane changes and shrinkage, independent of ceramide.

Area of Science:

  • Pharmacology
  • Cell Biology
  • Toxicology

Background:

  • FTY720 is a known apoptosis inducer in nucleated cells.
  • Eryptosis, or suicidal erythrocyte death, shares hallmarks with apoptosis.
  • Eryptosis is characterized by cell membrane scrambling and shrinkage, often due to increased intracellular calcium and ceramide.

Purpose of the Study:

  • To investigate if FTY720 induces eryptosis.
  • To determine the mechanisms by which FTY720 might affect erythrocytes.

Main Methods:

  • Annexin V-binding assay for cell membrane scrambling.
  • FACS analysis for cell shrinkage (forward scatter).
  • Fluo3 fluorescence for cytosolic Ca(2+) concentration.
  • Ceramide formation assessed via antibody binding.
  • Hemolysis measurement for hemoglobin release.

Main Results:

  • FTY720 (10 μM) significantly increased annexin V-binding and cytosolic Ca(2+) concentration within 48 hours.
  • FTY720 significantly decreased forward scatter, indicating cell shrinkage.
  • Ceramide formation was not significantly altered by FTY720 exposure.
  • The observed effects were significantly reduced when extracellular Ca(2+) was absent.

Conclusions:

  • FTY720 can induce suicidal erythrocyte death (eryptosis) at toxic concentrations.
  • The mechanism involves increased cytosolic Ca(2+) due to enhanced Ca(2+) entry.
  • FTY720-induced eryptosis is largely independent of ceramide accumulation.

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