Role of Extracellular Traps Promoted by Intestinal Parasites. Relationship with Virulence

Díaz-Godínez, César; Martínez-Flores, Arony; Argüello-García, Raúl; Olivos-García, Alfonso; Néquiz-Avendaño, Mario; Carrero, Julio César

doi:10.1007/978-3-030-44826-4_13

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“…This data suggests that some products of the E. histolytica trophozoites metabolism are responsible for inducing NETosis. In this context, ROS derived from pathogens have been identified as molecules that lead NET release independently of ROS produced by neutrophils [ 33 , 43 ]. Noteworthy, we previously showed that Entamoeba dispar , a non-pathogen human amoeba, does not produce ROS and does not trigger NETosis [ 43 , 47 ].…”

Section: Discussionmentioning

confidence: 99%

“…In this context, ROS derived from pathogens have been identified as molecules that lead NET release independently of ROS produced by neutrophils [ 33 , 43 ]. Noteworthy, we previously showed that Entamoeba dispar , a non-pathogen human amoeba, does not produce ROS and does not trigger NETosis [ 43 , 47 ]. Since most studies suggest that NETosis requires some source of ROS that does not come from neutrophils in this case, we considered the possibility that ROS produced by E. histolytica trophozoites induce NETosis.…”

Section: Discussionmentioning

confidence: 99%

“…NET quantitation was performed as described before [ 43 ]. In brief, neutrophils (5 × 10 5 ) were centrifuged at 4000 rpm for 2 min and resuspended in 500 µL of RPMI-1640 medium (Biological Industries) supplemented with 5% fetal bovine serum (FBS, Gibco) and 500 nM SYTOX ® Green (Invitrogen).…”

Section: Methodsmentioning

confidence: 99%

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New Insights on NETosis Induced by Entamoeba histolytica: Dependence on ROS from Amoebas and Extracellular MPO Activity

et al. 2021

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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.

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