Efavirenz Induced Suicidal Death of Human Erythrocytes

Rosi Bissinger , Ghada Bouguerra , Abdulla Al Mamun Bhuyan , Sabrina Waibel , Salem Abbes (1) , Florian Lang (2, 3)

1 Laboratory of Molecular and Cellular Hematology
2 Department of Physiology
3 Physiologisches Institut

Background/Aims: The reverse transcriptase inhibitor efavirenz utilized for the treatment of human immunodeficiency virus (HIV)-1 infection, triggers suicidal cell death or apoptosis, an effect in part due to interference with mitochondrial potential. Side effects of efavirenz include anemia. Causes of anemia include accelerated clearance of circulating erythrocytes. Even though lacking mitochondria, erythrocytes may enter suicidal erythrocyte death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine translocation to the erythrocyte surface. Triggers of eryptosis include Ca2+ entry and increase of cytosolic Ca2+ activity ([Ca2+](i)), oxidative stress, ceramide, as well as activation of p38 kinase, casein kinase 1 alpha and/or cyclooxygenase. The present study explored, whether and how efavirenz induces eryptosis. Methods: Phosphatidylserine exposure at the cell surface was estimated from annexin V binding, cell volume from forward scatter, [Ca2+](i) from Fluo3-fluorescence, ROS formation from DCFDA dependent fluorescence, and ceramide abundance utilizing selective antibodies. Results: A 48 hours exposure of human erythrocytes to efavirenz (>= 2 mu g/ml) significantly increased the percentage of annexin-V-binding cells, significantly decreased forward scatter (2 mu g/ml), significantly increased Fluo3-fluorescence (>= 2 mu g/ml), but did not significantly modify DCFDA fluorescence or ceramide abundance. The effect of efavirenz on annexin-V-binding was significantly blunted, but not abolished by removal of extracellular Ca2+. The effect of efavirenz on annexin-V-binding was further significantly blunted by p38 kinase inhibitor SB203580 (2 mu M) and casein kinase 1 alpha inhibitor D4476 (10 mu M), but not by cyclooxygenase inhibitor aspirin (50 mu M). Conclusions: Efavirenz triggers cell shrinkage and phosphatidylserine translocation to the erythrocyte surface, an effect in part due to stimulation of Ca2+ entry as well as activation of p38 kinase and casein kinase 1 alpha. (C) 2015 The Author(s) Published by S. Karger AG, Basel

Mots clés: Phosphatidylserine; Cell volume; Eryptosis; Oxidative stress; Calcium; p38 kinase; Casein kinase

Domaines: Sciences du Vivant [q-bio]

Rosi Bissinger, Ghada Bouguerra, Abdulla Al Mamun Bhuyan, Sabrina Waibel, Salem Abbes, et al.. Efavirenz Induced Suicidal Death of Human Erythrocytes. Cellular Physiology and Biochemistry, 2015, 37 (6), pp.2496-2507. ⟨10.1159/000438602⟩. ⟨hal-01358496⟩