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A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses

Trichinella spiralis can modulate host immune responses to retain a suitable environment for its long-term survival. Incidentally, the parasite elicits regulatory effects through immunomodulatory molecule release, which can suppress host inflammation and may be used for the treatment of unrelated in...

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Autores principales: Kobpornchai, Porntida, Flynn, Robin J., Reamtong, Onrapak, Rittisoonthorn, Nonglucksanawan, Kosoltanapiwat, Nathamon, Boonnak, Kobporn, Boonyuen, Usa, Ampawong, Sumate, Jiratanh, Montakan, Tattiyapong, Muncharee, Adisakwattana, Poom
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7153903/
https://www.ncbi.nlm.nih.gov/pubmed/32236093
http://dx.doi.org/10.1371/journal.pntd.0008192
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author Kobpornchai, Porntida
Flynn, Robin J.
Reamtong, Onrapak
Rittisoonthorn, Nonglucksanawan
Kosoltanapiwat, Nathamon
Boonnak, Kobporn
Boonyuen, Usa
Ampawong, Sumate
Jiratanh, Montakan
Tattiyapong, Muncharee
Adisakwattana, Poom
author_facet Kobpornchai, Porntida
Flynn, Robin J.
Reamtong, Onrapak
Rittisoonthorn, Nonglucksanawan
Kosoltanapiwat, Nathamon
Boonnak, Kobporn
Boonyuen, Usa
Ampawong, Sumate
Jiratanh, Montakan
Tattiyapong, Muncharee
Adisakwattana, Poom
author_sort Kobpornchai, Porntida
collection PubMed
description Trichinella spiralis can modulate host immune responses to retain a suitable environment for its long-term survival. Incidentally, the parasite elicits regulatory effects through immunomodulatory molecule release, which can suppress host inflammation and may be used for the treatment of unrelated inflammatory diseases in someday. Here we identified and characterized a novel T. spiralis cystatin (TsCstN), which inhibits inflammation mediated by LPS-treated macrophages.Proteins contained in the excretory–secretory (ES) product of muscle-stage T. spiralis (ES-L1) were fractionated, and each was treated with mouse bone marrow-derived macrophages (mBMDMs) before LPS stimulation. The fractions that exhibited high immunomodulatory property by decreasing pro-inflammatory cytokines or increasing anti-inflammatory cytokines were identified by mass spectrometry. Incidentally, the conserved hypothetical protein (Tsp_04814) was selected for further characterization as it presented the most significant MS score. An annotation of Tsp_04814 using protein structural homology comparison suggested that it has high structural similarity to human cystatin E/M (TM score 0.690). The recombinant T. spiralis novel cystatin (rTsCstN) was expressed in Escherichia coli at a molecular weight of approximately 13 kDa. Mouse anti-rTsCstN polyclonal antibody (pAb) could detect native TsCstN in crude worm antigens (CWA) and ES-L1 and be predominantly localized in the stichosome and subcuticular cells. rTsCstN inhibited cysteine proteases in vitro, especially cathepsin L, at an optimal pH of 6. Besides, rTsCstN could be internalized into mBMDMs, which were mostly distributed in the cytoplasm and lysosome both before and after LPS stimulation. To evaluate the rTsCstN immunomodulatory properties on mBMDMs, rTsCstN was incubated with mBMDM before LPS stimulation; this demonstrated that rTsCstN suppressed pro-inflammatory cytokine production and MHC class II expression.T. spiralis L1-derived TsCstN was characterized as a novel cysteine protease inhibitor. The protein elicits an anti-inflammatory property by suppressing pro-inflammatory cytokines and interfering with the antigen presentation process through depletion of MHC class II expression.
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spelling pubmed-71539032020-04-24 A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses Kobpornchai, Porntida Flynn, Robin J. Reamtong, Onrapak Rittisoonthorn, Nonglucksanawan Kosoltanapiwat, Nathamon Boonnak, Kobporn Boonyuen, Usa Ampawong, Sumate Jiratanh, Montakan Tattiyapong, Muncharee Adisakwattana, Poom PLoS Negl Trop Dis Research Article Trichinella spiralis can modulate host immune responses to retain a suitable environment for its long-term survival. Incidentally, the parasite elicits regulatory effects through immunomodulatory molecule release, which can suppress host inflammation and may be used for the treatment of unrelated inflammatory diseases in someday. Here we identified and characterized a novel T. spiralis cystatin (TsCstN), which inhibits inflammation mediated by LPS-treated macrophages.Proteins contained in the excretory–secretory (ES) product of muscle-stage T. spiralis (ES-L1) were fractionated, and each was treated with mouse bone marrow-derived macrophages (mBMDMs) before LPS stimulation. The fractions that exhibited high immunomodulatory property by decreasing pro-inflammatory cytokines or increasing anti-inflammatory cytokines were identified by mass spectrometry. Incidentally, the conserved hypothetical protein (Tsp_04814) was selected for further characterization as it presented the most significant MS score. An annotation of Tsp_04814 using protein structural homology comparison suggested that it has high structural similarity to human cystatin E/M (TM score 0.690). The recombinant T. spiralis novel cystatin (rTsCstN) was expressed in Escherichia coli at a molecular weight of approximately 13 kDa. Mouse anti-rTsCstN polyclonal antibody (pAb) could detect native TsCstN in crude worm antigens (CWA) and ES-L1 and be predominantly localized in the stichosome and subcuticular cells. rTsCstN inhibited cysteine proteases in vitro, especially cathepsin L, at an optimal pH of 6. Besides, rTsCstN could be internalized into mBMDMs, which were mostly distributed in the cytoplasm and lysosome both before and after LPS stimulation. To evaluate the rTsCstN immunomodulatory properties on mBMDMs, rTsCstN was incubated with mBMDM before LPS stimulation; this demonstrated that rTsCstN suppressed pro-inflammatory cytokine production and MHC class II expression.T. spiralis L1-derived TsCstN was characterized as a novel cysteine protease inhibitor. The protein elicits an anti-inflammatory property by suppressing pro-inflammatory cytokines and interfering with the antigen presentation process through depletion of MHC class II expression. Public Library of Science 2020-04-01 /pmc/articles/PMC7153903/ /pubmed/32236093 http://dx.doi.org/10.1371/journal.pntd.0008192 Text en © 2020 Kobpornchai et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Kobpornchai, Porntida
Flynn, Robin J.
Reamtong, Onrapak
Rittisoonthorn, Nonglucksanawan
Kosoltanapiwat, Nathamon
Boonnak, Kobporn
Boonyuen, Usa
Ampawong, Sumate
Jiratanh, Montakan
Tattiyapong, Muncharee
Adisakwattana, Poom
A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses
title A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses
title_full A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses
title_fullStr A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses
title_full_unstemmed A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses
title_short A novel cystatin derived from Trichinella spiralis suppresses macrophage-mediated inflammatory responses
title_sort novel cystatin derived from trichinella spiralis suppresses macrophage-mediated inflammatory responses
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7153903/
https://www.ncbi.nlm.nih.gov/pubmed/32236093
http://dx.doi.org/10.1371/journal.pntd.0008192
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