Shiga Toxin 2a Induces NETosis via NOX-Dependent Pathway.
Wouter J C Feitz, Samuel Suntharalingham, Meraj Khan, Carolina G Ortiz-Sandoval, Nades Palaniyar, Lambert P van den Heuvel, Nicole C A J van de Kar, Christoph Licht
Author Information
Wouter J C Feitz: Department of Pediatric Nephrology, Amalia Children's Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The Netherlands.
Samuel Suntharalingham: Cell Biology Program, The Hospital for Sick Children Research Institute, Toronto, ON M5G 1X8, Canada.
Meraj Khan: Program in Translational Medicine, The Hospital for Sick Children Research Institute, Toronto, ON M5G 1X8, Canada.
Carolina G Ortiz-Sandoval: Cell Biology Program, The Hospital for Sick Children Research Institute, Toronto, ON M5G 1X8, Canada.
Nades Palaniyar: Program in Translational Medicine, The Hospital for Sick Children Research Institute, Toronto, ON M5G 1X8, Canada. ORCID
Lambert P van den Heuvel: Department of Pediatric Nephrology, Amalia Children's Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The Netherlands.
Nicole C A J van de Kar: Department of Pediatric Nephrology, Amalia Children's Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The Netherlands. ORCID
Christoph Licht: Cell Biology Program, The Hospital for Sick Children Research Institute, Toronto, ON M5G 1X8, Canada. ORCID
Shiga toxin (Stx)-producing (STEC) infection is the most common cause of hemolytic uremic syndrome (HUS), one of the main causes of acute kidney injury in children. Stx plays an important role in endothelium damage and pathogenesis of STEC-HUS. However, the effects of Stx on neutrophils and neutrophil extracellular trap (NET) formation are not well understood. In this study, we investigated how Stx2a affects NET formation and NETotic pathways (NADPH or NOX-dependent and -independent) using neutrophils isolated from healthy donors and patients with STEC-HUS, during the acute and recovery phase of the disease. Stx2a dose-dependently induced NETosis in neutrophils isolated from both healthy controls and STEC-HUSpatients. NETosis kinetics and mechanistic data with pathway-specific inhibitors including diphenyleneiodonium (DPI)-, ERK-, and P38-inhibitors showed that Stx2a-induced NETosis via the NOX-dependent pathway. Neutrophils from STEC-HUSpatients in the acute phase showed less ROS and NETs formation compared to neutrophils of the recovery phase of the disease and in healthy controls. NETs induced by Stx2a may lead to the activation of endothelial cells, which might contribute to the manifestation of thrombotic microangiopathy in STEC-HUS.
