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Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils

Visceral leishmaniasis (VL) is often associated with hematologic manifestations that may interfere with neutrophil response. Lipophosphoglycan (LPG) is a major molecule on the surface of Leishmania promastigotes, which has been associated with several aspects of the parasite–vector–host interplay. H...

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Autores principales: Quintela-Carvalho, Graziele, Goicochea, Astrid Madeleine Calero, Mançur-Santos, Vanessa, Viana, Sayonara de Melo, Luz, Yasmin da Silva, Dias, Beatriz Rocha Simões, Lázaro-Souza, Milena, Suarez, Martha, de Oliveira, Camila Indiani, Saraiva, Elvira M., Brodskyn, Cláudia I., Veras, Patrícia T., de Menezes, Juliana P.B., Andrade, Bruno B., Lima, Jonilson Berlink, Descoteaux, Albert, Borges, Valéria M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9019130/
https://www.ncbi.nlm.nih.gov/pubmed/35463648
http://dx.doi.org/10.3389/fcimb.2022.788196
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author Quintela-Carvalho, Graziele
Goicochea, Astrid Madeleine Calero
Mançur-Santos, Vanessa
Viana, Sayonara de Melo
Luz, Yasmin da Silva
Dias, Beatriz Rocha Simões
Lázaro-Souza, Milena
Suarez, Martha
de Oliveira, Camila Indiani
Saraiva, Elvira M.
Brodskyn, Cláudia I.
Veras, Patrícia T.
de Menezes, Juliana P.B.
Andrade, Bruno B.
Lima, Jonilson Berlink
Descoteaux, Albert
Borges, Valéria M.
author_facet Quintela-Carvalho, Graziele
Goicochea, Astrid Madeleine Calero
Mançur-Santos, Vanessa
Viana, Sayonara de Melo
Luz, Yasmin da Silva
Dias, Beatriz Rocha Simões
Lázaro-Souza, Milena
Suarez, Martha
de Oliveira, Camila Indiani
Saraiva, Elvira M.
Brodskyn, Cláudia I.
Veras, Patrícia T.
de Menezes, Juliana P.B.
Andrade, Bruno B.
Lima, Jonilson Berlink
Descoteaux, Albert
Borges, Valéria M.
author_sort Quintela-Carvalho, Graziele
collection PubMed
description Visceral leishmaniasis (VL) is often associated with hematologic manifestations that may interfere with neutrophil response. Lipophosphoglycan (LPG) is a major molecule on the surface of Leishmania promastigotes, which has been associated with several aspects of the parasite–vector–host interplay. Here, we investigated how LPG from Leishmania (L.) infantum, the principal etiological agent of VL in the New World, influences the initial establishment of infection during interaction with human neutrophils in an experimental setting in vitro. Human neutrophils obtained from peripheral blood samples were infected with either the wild-type L. infantum (WT) strain or LPG-deficient mutant (∆lpg1). In this setting, ∆lpg1 parasites displayed reduced viability compared to WT L. infantum; such finding was reverted in the complemented ∆lpg1+LPG1 parasites at 3- and 6-h post-infection. Confocal microscopy experiments indicated that this decreased survival was related to enhanced lysosomal fusion. In fact, LPG-deficient L. infantum parasites more frequently died inside neutrophil acidic compartments, a phenomenon that was reverted when host cells were treated with Wortmannin. We also observed an increase in the secretion of the neutrophil collagenase matrix metalloproteinase-8 (MMP-8) by cells infected with ∆lpg1 L. infantum compared to those that were infected with WT parasites. Furthermore, collagen I matrix degradation was found to be significantly increased in ∆lpg1 parasite-infected cells but not in WT-infected controls. Flow cytometry analysis revealed a substantial boost in production of reactive oxygen species (ROS) during infection with either WT or ∆lpg1 L. infantum. In addition, killing of ∆lpg1 parasites was shown to be more dependent on the ROS production than that of WT L. infantum. Notably, inhibition of the oxidative stress with Apocynin potentially fueled ∆lpg1 L. infantum fitness as it increased the intracellular parasite viability. Thus, our observations demonstrate that LPG may be a critical molecule fostering parasite survival in human neutrophils through a mechanism that involves cellular activation and generation of free radicals.
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spelling pubmed-90191302022-04-21 Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils Quintela-Carvalho, Graziele Goicochea, Astrid Madeleine Calero Mançur-Santos, Vanessa Viana, Sayonara de Melo Luz, Yasmin da Silva Dias, Beatriz Rocha Simões Lázaro-Souza, Milena Suarez, Martha de Oliveira, Camila Indiani Saraiva, Elvira M. Brodskyn, Cláudia I. Veras, Patrícia T. de Menezes, Juliana P.B. Andrade, Bruno B. Lima, Jonilson Berlink Descoteaux, Albert Borges, Valéria M. Front Cell Infect Microbiol Cellular and Infection Microbiology Visceral leishmaniasis (VL) is often associated with hematologic manifestations that may interfere with neutrophil response. Lipophosphoglycan (LPG) is a major molecule on the surface of Leishmania promastigotes, which has been associated with several aspects of the parasite–vector–host interplay. Here, we investigated how LPG from Leishmania (L.) infantum, the principal etiological agent of VL in the New World, influences the initial establishment of infection during interaction with human neutrophils in an experimental setting in vitro. Human neutrophils obtained from peripheral blood samples were infected with either the wild-type L. infantum (WT) strain or LPG-deficient mutant (∆lpg1). In this setting, ∆lpg1 parasites displayed reduced viability compared to WT L. infantum; such finding was reverted in the complemented ∆lpg1+LPG1 parasites at 3- and 6-h post-infection. Confocal microscopy experiments indicated that this decreased survival was related to enhanced lysosomal fusion. In fact, LPG-deficient L. infantum parasites more frequently died inside neutrophil acidic compartments, a phenomenon that was reverted when host cells were treated with Wortmannin. We also observed an increase in the secretion of the neutrophil collagenase matrix metalloproteinase-8 (MMP-8) by cells infected with ∆lpg1 L. infantum compared to those that were infected with WT parasites. Furthermore, collagen I matrix degradation was found to be significantly increased in ∆lpg1 parasite-infected cells but not in WT-infected controls. Flow cytometry analysis revealed a substantial boost in production of reactive oxygen species (ROS) during infection with either WT or ∆lpg1 L. infantum. In addition, killing of ∆lpg1 parasites was shown to be more dependent on the ROS production than that of WT L. infantum. Notably, inhibition of the oxidative stress with Apocynin potentially fueled ∆lpg1 L. infantum fitness as it increased the intracellular parasite viability. Thus, our observations demonstrate that LPG may be a critical molecule fostering parasite survival in human neutrophils through a mechanism that involves cellular activation and generation of free radicals. Frontiers Media S.A. 2022-04-06 /pmc/articles/PMC9019130/ /pubmed/35463648 http://dx.doi.org/10.3389/fcimb.2022.788196 Text en Copyright © 2022 Quintela-Carvalho, Goicochea, Mançur-Santos, Viana, Luz, Dias, Lázaro-Souza, Suarez, de Oliveira, Saraiva, Brodskyn, Veras, de Menezes, Andrade, Lima, Descoteaux and Borges https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Quintela-Carvalho, Graziele
Goicochea, Astrid Madeleine Calero
Mançur-Santos, Vanessa
Viana, Sayonara de Melo
Luz, Yasmin da Silva
Dias, Beatriz Rocha Simões
Lázaro-Souza, Milena
Suarez, Martha
de Oliveira, Camila Indiani
Saraiva, Elvira M.
Brodskyn, Cláudia I.
Veras, Patrícia T.
de Menezes, Juliana P.B.
Andrade, Bruno B.
Lima, Jonilson Berlink
Descoteaux, Albert
Borges, Valéria M.
Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils
title Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils
title_full Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils
title_fullStr Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils
title_full_unstemmed Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils
title_short Leishmania infantum Defective in Lipophosphoglycan Biosynthesis Interferes With Activation of Human Neutrophils
title_sort leishmania infantum defective in lipophosphoglycan biosynthesis interferes with activation of human neutrophils
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9019130/
https://www.ncbi.nlm.nih.gov/pubmed/35463648
http://dx.doi.org/10.3389/fcimb.2022.788196
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