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l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis

The survival of intracellular protozoan parasite, Leishmania donovani, the causative agent of Indian visceral leishmaniasis (VL), depends on the activation status of macrophages. l-Arginine, a semi-essential amino acid plays a crucial regulatory role for activation of macrophages. However, the role...

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Autores principales: Mandal, Abhishek, Das, Sushmita, Kumar, Ajay, Roy, Saptarshi, Verma, Sudha, Ghosh, Ayan Kumar, Singh, Ruby, Abhishek, Kumar, Saini, Savita, Sardar, Abul Hasan, Purkait, Bidyut, Kumar, Ashish, Mandal, Chitra, Das, Pradeep
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5526900/
https://www.ncbi.nlm.nih.gov/pubmed/28798743
http://dx.doi.org/10.3389/fimmu.2017.00839
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author Mandal, Abhishek
Das, Sushmita
Kumar, Ajay
Roy, Saptarshi
Verma, Sudha
Ghosh, Ayan Kumar
Singh, Ruby
Abhishek, Kumar
Saini, Savita
Sardar, Abul Hasan
Purkait, Bidyut
Kumar, Ashish
Mandal, Chitra
Das, Pradeep
author_facet Mandal, Abhishek
Das, Sushmita
Kumar, Ajay
Roy, Saptarshi
Verma, Sudha
Ghosh, Ayan Kumar
Singh, Ruby
Abhishek, Kumar
Saini, Savita
Sardar, Abul Hasan
Purkait, Bidyut
Kumar, Ashish
Mandal, Chitra
Das, Pradeep
author_sort Mandal, Abhishek
collection PubMed
description The survival of intracellular protozoan parasite, Leishmania donovani, the causative agent of Indian visceral leishmaniasis (VL), depends on the activation status of macrophages. l-Arginine, a semi-essential amino acid plays a crucial regulatory role for activation of macrophages. However, the role of l-arginine transport in VL still remains elusive. In this study, we demonstrated that intra-macrophage survival of L. donovani depends on the availability of extracellular l-arginine. Infection of THP-1-derived macrophage/human monocyte-derived macrophage (hMDM) with Leishmania, resulted in upregulation of l-arginine transport. While investigating the involvement of the transporters, we observed that Leishmania survival was greatly impaired when the transporters were blocked either using inhibitor or siRNA-mediated downregulation. CAT-2 was found to be the main isoform associated with l-arginine transport in L. donovani-infected macrophages. l-arginine availability and its transport regulated the host arginase in Leishmania infection. Arginase and inducible nitric oxide synthase (iNOS) expression were reciprocally regulated when assayed using specific inhibitors and siRNA-mediated downregulation. Interestingly, induction of iNOS expression and nitric oxide production were observed in case of inhibition of arginase in infected macrophages. Furthermore, inhibition of l-arginine transport as well as arginase resulted in decreased polyamine production, limiting parasite survival inside macrophages. l-arginine availability and transport regulated Th1/Th2 cytokine levels in case of Leishmania infection. Upregulation of l-arginine transport, induction of host arginase, and enhanced polyamine production were correlated with increased level of IL-10 and decreased level of IL-12 and TNF-α in L. donovani-infected macrophages. Our findings provide clear evidence for targeting the metabolism of l-arginine and l-arginine-metabolizing enzymes as an important therapeutic and prophylactic strategy to treat VL.
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spelling pubmed-55269002017-08-10 l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis Mandal, Abhishek Das, Sushmita Kumar, Ajay Roy, Saptarshi Verma, Sudha Ghosh, Ayan Kumar Singh, Ruby Abhishek, Kumar Saini, Savita Sardar, Abul Hasan Purkait, Bidyut Kumar, Ashish Mandal, Chitra Das, Pradeep Front Immunol Immunology The survival of intracellular protozoan parasite, Leishmania donovani, the causative agent of Indian visceral leishmaniasis (VL), depends on the activation status of macrophages. l-Arginine, a semi-essential amino acid plays a crucial regulatory role for activation of macrophages. However, the role of l-arginine transport in VL still remains elusive. In this study, we demonstrated that intra-macrophage survival of L. donovani depends on the availability of extracellular l-arginine. Infection of THP-1-derived macrophage/human monocyte-derived macrophage (hMDM) with Leishmania, resulted in upregulation of l-arginine transport. While investigating the involvement of the transporters, we observed that Leishmania survival was greatly impaired when the transporters were blocked either using inhibitor or siRNA-mediated downregulation. CAT-2 was found to be the main isoform associated with l-arginine transport in L. donovani-infected macrophages. l-arginine availability and its transport regulated the host arginase in Leishmania infection. Arginase and inducible nitric oxide synthase (iNOS) expression were reciprocally regulated when assayed using specific inhibitors and siRNA-mediated downregulation. Interestingly, induction of iNOS expression and nitric oxide production were observed in case of inhibition of arginase in infected macrophages. Furthermore, inhibition of l-arginine transport as well as arginase resulted in decreased polyamine production, limiting parasite survival inside macrophages. l-arginine availability and transport regulated Th1/Th2 cytokine levels in case of Leishmania infection. Upregulation of l-arginine transport, induction of host arginase, and enhanced polyamine production were correlated with increased level of IL-10 and decreased level of IL-12 and TNF-α in L. donovani-infected macrophages. Our findings provide clear evidence for targeting the metabolism of l-arginine and l-arginine-metabolizing enzymes as an important therapeutic and prophylactic strategy to treat VL. Frontiers Media S.A. 2017-07-26 /pmc/articles/PMC5526900/ /pubmed/28798743 http://dx.doi.org/10.3389/fimmu.2017.00839 Text en Copyright © 2017 Mandal, Das, Kumar, Roy, Verma, Ghosh, Singh, Abhishek, Saini, Sardar, Purkait, Kumar, Mandal and Das. http://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) or licensor 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 Immunology
Mandal, Abhishek
Das, Sushmita
Kumar, Ajay
Roy, Saptarshi
Verma, Sudha
Ghosh, Ayan Kumar
Singh, Ruby
Abhishek, Kumar
Saini, Savita
Sardar, Abul Hasan
Purkait, Bidyut
Kumar, Ashish
Mandal, Chitra
Das, Pradeep
l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis
title l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis
title_full l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis
title_fullStr l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis
title_full_unstemmed l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis
title_short l-Arginine Uptake by Cationic Amino Acid Transporter Promotes Intra-Macrophage Survival of Leishmania donovani by Enhancing Arginase-Mediated Polyamine Synthesis
title_sort l-arginine uptake by cationic amino acid transporter promotes intra-macrophage survival of leishmania donovani by enhancing arginase-mediated polyamine synthesis
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5526900/
https://www.ncbi.nlm.nih.gov/pubmed/28798743
http://dx.doi.org/10.3389/fimmu.2017.00839
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