New Insights on NETosis Induced by Entamoeba histolytica: Dependence on ROS from Amoebas and Extracellular MPO Activity
Por:
Diaz-Godinez, Cesar, Jorge-Rosas, Joshue Fabian, Nequiz, Mario, Martinez-Calvillo, Santiago, Laclette, Juan P., Rosales, Carlos, Carrero, Julio C.
Publicada:
1 jun 2021
Resumen:
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(R) 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.
Filiaciones:
Diaz-Godinez, Cesar:
Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, 04510, Mexico
Jorge-Rosas, Joshue Fabian:
Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, 04510, Mexico
Nequiz, Mario:
Unidad de Investigación en Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, Ciudad de México, 06720, Mexico
Martinez-Calvillo, Santiago:
Unidad de Biomedicina, Facultad de Estudios Superiores Iztacala, Universidad Nacional Autónoma de México, Av. de los Barrios 1, Col. Los Reyes Iztacala, Tlalnepantla, Estado de México, 54090, Mexico
Laclette, Juan P.:
Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, 04510, Mexico
Rosales, Carlos:
Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, 04510, Mexico
Carrero, Julio C.:
Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México (UNAM), Ciudad de México, 04510, Mexico
Green Published, gold, Gold
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