Cargando…

Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production

Myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA)-associated vasculitis is a systemic small-vessel vasculitis, wherein, MPO-ANCA plays a critical role in the pathogenesis. Neutrophil extracellular traps (NETs) released from activated neutrophils are composed of extracellular web-like DN...

Descripción completa

Detalles Bibliográficos
Autores principales: Kusunoki, Yoshihiro, Nakazawa, Daigo, Shida, Haruki, Hattanda, Fumihiko, Miyoshi, Arina, Masuda, Sakiko, Nishio, Saori, Tomaru, Utano, Atsumi, Tatsuya, Ishizu, Akihiro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4896908/
https://www.ncbi.nlm.nih.gov/pubmed/27375623
http://dx.doi.org/10.3389/fimmu.2016.00227
_version_ 1782436044217516032
author Kusunoki, Yoshihiro
Nakazawa, Daigo
Shida, Haruki
Hattanda, Fumihiko
Miyoshi, Arina
Masuda, Sakiko
Nishio, Saori
Tomaru, Utano
Atsumi, Tatsuya
Ishizu, Akihiro
author_facet Kusunoki, Yoshihiro
Nakazawa, Daigo
Shida, Haruki
Hattanda, Fumihiko
Miyoshi, Arina
Masuda, Sakiko
Nishio, Saori
Tomaru, Utano
Atsumi, Tatsuya
Ishizu, Akihiro
author_sort Kusunoki, Yoshihiro
collection PubMed
description Myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA)-associated vasculitis is a systemic small-vessel vasculitis, wherein, MPO-ANCA plays a critical role in the pathogenesis. Neutrophil extracellular traps (NETs) released from activated neutrophils are composed of extracellular web-like DNA and antimicrobial proteins, including MPO. Diverse stimuli, such as phorbol myristate acetate (PMA) and ligands of toll-like receptors (TLR), induce NETs. Although TLR-mediated NET formation can occur with preservation of living neutrophilic functions (called vital NETosis), PMA-stimulated neutrophils undergo cell death with NET formation (called suicidal NETosis). In the process of suicidal NETosis, histones are citrullinated by peptidylarginine deiminase 4 (PAD4). Since this step is necessary for decondensation of DNA, PAD4 plays a pivotal role in suicidal NETosis. Although NETs are essential for elimination of microorganisms, excessive formation of NETs has been suggested to be implicated in MPO-ANCA production. This study aimed to determine if pan-PAD inhibitors could suppress MPO-ANCA production in vivo. At first, NETs were induced in peripheral blood neutrophils derived from healthy donors (1 × 10(6)/ml) by stimulation with 20 nM PMA with or without 20 μM propylthiouracil (PTU), an anti-thyroid drug. We then determined that the in vitro NET formation was inhibited completely by 200 μM Cl-amidine, a pan-PAD inhibitor. Next, we established mouse models with MPO-ANCA production. BALB/c mice were given intraperitoneal (i.p.) injection of PMA (50 ng at days 0 and 7) and oral PTU (2.5 mg/day) for 2 weeks. These mice were divided into two groups; the first group was given daily i.p. injection of PBS (200 μl/day) (n = 13) and the other group with daily i.p. injection of Cl-amidine (0.3 mg/200 μl PBS/day) (n = 7). Two weeks later, citrullination as an indicator of NET formation in the peritoneum and serum MPO-ANCA titer was compared between the two groups. Results demonstrated that citrullination in the peritoneum was significantly reduced in the Cl-amidine-treated mice compared with the vehicle-injected control mice (38% reduction). Additionally, the serum MPO-ANCA titer of the Cl-amidine-treated mice (32.3 ± 31.0 ng/ml) was significantly lower than that in the vehicle-injected mice (132.1 ± 41.6 ng/ml). The collective findings indicate that excessive formation of NETs may be implicated in MPO-ANCA production in vivo.
format Online
Article
Text
id pubmed-4896908
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-48969082016-07-01 Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production Kusunoki, Yoshihiro Nakazawa, Daigo Shida, Haruki Hattanda, Fumihiko Miyoshi, Arina Masuda, Sakiko Nishio, Saori Tomaru, Utano Atsumi, Tatsuya Ishizu, Akihiro Front Immunol Immunology Myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA)-associated vasculitis is a systemic small-vessel vasculitis, wherein, MPO-ANCA plays a critical role in the pathogenesis. Neutrophil extracellular traps (NETs) released from activated neutrophils are composed of extracellular web-like DNA and antimicrobial proteins, including MPO. Diverse stimuli, such as phorbol myristate acetate (PMA) and ligands of toll-like receptors (TLR), induce NETs. Although TLR-mediated NET formation can occur with preservation of living neutrophilic functions (called vital NETosis), PMA-stimulated neutrophils undergo cell death with NET formation (called suicidal NETosis). In the process of suicidal NETosis, histones are citrullinated by peptidylarginine deiminase 4 (PAD4). Since this step is necessary for decondensation of DNA, PAD4 plays a pivotal role in suicidal NETosis. Although NETs are essential for elimination of microorganisms, excessive formation of NETs has been suggested to be implicated in MPO-ANCA production. This study aimed to determine if pan-PAD inhibitors could suppress MPO-ANCA production in vivo. At first, NETs were induced in peripheral blood neutrophils derived from healthy donors (1 × 10(6)/ml) by stimulation with 20 nM PMA with or without 20 μM propylthiouracil (PTU), an anti-thyroid drug. We then determined that the in vitro NET formation was inhibited completely by 200 μM Cl-amidine, a pan-PAD inhibitor. Next, we established mouse models with MPO-ANCA production. BALB/c mice were given intraperitoneal (i.p.) injection of PMA (50 ng at days 0 and 7) and oral PTU (2.5 mg/day) for 2 weeks. These mice were divided into two groups; the first group was given daily i.p. injection of PBS (200 μl/day) (n = 13) and the other group with daily i.p. injection of Cl-amidine (0.3 mg/200 μl PBS/day) (n = 7). Two weeks later, citrullination as an indicator of NET formation in the peritoneum and serum MPO-ANCA titer was compared between the two groups. Results demonstrated that citrullination in the peritoneum was significantly reduced in the Cl-amidine-treated mice compared with the vehicle-injected control mice (38% reduction). Additionally, the serum MPO-ANCA titer of the Cl-amidine-treated mice (32.3 ± 31.0 ng/ml) was significantly lower than that in the vehicle-injected mice (132.1 ± 41.6 ng/ml). The collective findings indicate that excessive formation of NETs may be implicated in MPO-ANCA production in vivo. Frontiers Media S.A. 2016-06-08 /pmc/articles/PMC4896908/ /pubmed/27375623 http://dx.doi.org/10.3389/fimmu.2016.00227 Text en Copyright © 2016 Kusunoki, Nakazawa, Shida, Hattanda, Miyoshi, Masuda, Nishio, Tomaru, Atsumi and Ishizu. 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
Kusunoki, Yoshihiro
Nakazawa, Daigo
Shida, Haruki
Hattanda, Fumihiko
Miyoshi, Arina
Masuda, Sakiko
Nishio, Saori
Tomaru, Utano
Atsumi, Tatsuya
Ishizu, Akihiro
Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
title Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
title_full Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
title_fullStr Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
title_full_unstemmed Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
title_short Peptidylarginine Deiminase Inhibitor Suppresses Neutrophil Extracellular Trap Formation and MPO-ANCA Production
title_sort peptidylarginine deiminase inhibitor suppresses neutrophil extracellular trap formation and mpo-anca production
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4896908/
https://www.ncbi.nlm.nih.gov/pubmed/27375623
http://dx.doi.org/10.3389/fimmu.2016.00227
work_keys_str_mv AT kusunokiyoshihiro peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT nakazawadaigo peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT shidaharuki peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT hattandafumihiko peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT miyoshiarina peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT masudasakiko peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT nishiosaori peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT tomaruutano peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT atsumitatsuya peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction
AT ishizuakihiro peptidylargininedeiminaseinhibitorsuppressesneutrophilextracellulartrapformationandmpoancaproduction