New Insights on NETosis Induced by Entamoeba histolytica: Dependence on ROS from Amoebas and Extracellular MPO Activity
Open Access
- 18 June 2021
- journal article
- research article
- Published by MDPI AG in Antioxidants
- Vol. 10 (6), 974
- https://doi.org/10.3390/antiox10060974
Abstract
NETosis is a neutrophil process involving sequential steps from pathogen detection to the release of DNA harboring antimicrobial proteins, including the central generation of NADPH oxidase dependent or independent ROS. Previously, we reported that NETosis triggered by Entamoeba histolytica trophozoites is independent of NADPH oxidase activity in neutrophils, but dependent on the viability of the parasites and no ROS source was identified. Here, we explored the possibility that E. histolytica trophozoites serve as the ROS source for NETosis. NET quantitation was performed using SYTOX® Green assay in the presence of selective inhibitors and scavengers. We observed that respiratory burst in neutrophils was inhibited by trophozoites in a dose dependent manner. Mitochondrial ROS was not also necessary, as the mitochondrial scavenger mitoTEMPO did not affect the process. Surprisingly, ROS-deficient amoebas obtained by pre-treatment with pyrocatechol were less likely to induce NETs. Additionally, we detected the presence of MPO on the cell surface of trophozoites after the interaction with neutrophils and found that luminol and isoluminol, intracellular and extracellular scavengers for MPO derived ROS reduced the amount of NET triggered by amoebas. These data suggest that ROS generated by trophozoites and processed by the extracellular MPO during the contact with neutrophils are required for E. histolytica induced NETosis.Funding Information
- Consejo Nacional de Ciencia y Tecnología (284830 (to J.C.C.) and 254434 (to C.R.))
- Programa de Apoyo a Proyectos de Investigación e Innovación Tecnológica (IN208020 (to J.C.C.))
This publication has 88 references indexed in Scilit:
- Toxoplasma gondii Triggers Release of Human and Mouse Neutrophil Extracellular TrapsInfection and Immunity, 2012
- Hypochlorous acid regulates neutrophil extracellular trap release in humansClinical and Experimental Immunology, 2011
- Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular trapsThe Journal of cell biology, 2010
- PAD4 is essential for antibacterial innate immunity mediated by neutrophil extracellular trapsThe Journal of Experimental Medicine, 2010
- Optimization of Formaldehyde Cross-Linking for Protein Interaction Analysis of Non-Tagged Integrin1Journal of Biomedicine and Biotechnology, 2010
- Leishmania amazonensispromastigotes induce and are killed by neutrophil extracellular trapsProceedings of the National Academy of Sciences of the United States of America, 2009
- Bioluminescence imaging of myeloperoxidase activity in vivoNature Medicine, 2009
- Histone hypercitrullination mediates chromatin decondensation and neutrophil extracellular trap formationThe Journal of cell biology, 2009
- Novel cell death program leads to neutrophil extracellular trapsThe Journal of cell biology, 2007
- Neutrophil Extracellular Traps Kill BacteriaScience, 2004