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Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils
BACKGROUND: Granulocyte–macrophage colony-stimulating factor (GM-CSF) has emerged as a crucial cytokine that activates myeloid cells to initiate tissue inflammation. However, the molecular actions of GM-CSF against innate immunity are still poorly characterized. Here, we investigated the in vitro ef...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6116484/ https://www.ncbi.nlm.nih.gov/pubmed/30157949 http://dx.doi.org/10.1186/s13075-018-1685-x |
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author | Furuya, Makiko Yashiro Asano, Tomoyuki Sumichika, Yuya Sato, Shuzo Kobayashi, Hiroko Watanabe, Hiroshi Suzuki, Eiji Kozuru, Hideko Yatsuhashi, Hiroshi Koga, Tomohiro Ohira, Hiromasa Sekine, Hideharu Kawakami, Atsushi Migita, Kiyoshi |
author_facet | Furuya, Makiko Yashiro Asano, Tomoyuki Sumichika, Yuya Sato, Shuzo Kobayashi, Hiroko Watanabe, Hiroshi Suzuki, Eiji Kozuru, Hideko Yatsuhashi, Hiroshi Koga, Tomohiro Ohira, Hiromasa Sekine, Hideharu Kawakami, Atsushi Migita, Kiyoshi |
author_sort | Furuya, Makiko Yashiro |
collection | PubMed |
description | BACKGROUND: Granulocyte–macrophage colony-stimulating factor (GM-CSF) has emerged as a crucial cytokine that activates myeloid cells to initiate tissue inflammation. However, the molecular actions of GM-CSF against innate immunity are still poorly characterized. Here, we investigated the in vitro effects of GM-CSF on the activation of human myeloid lineages, neutrophils, and the underlying intracellular signaling mechanism, including inflammasome activation. METHODS: Human neutrophils were stimulated with GM-CSF in the presence or absence of tofacitinib. The cellular supernatants were analyzed for interleukin-1 beta (IL-1β) and caspase-1 by enzyme-linked immunosorbent assay (ELISA) methods. Pro-IL-1β mRNA expressions in human neutrophils were analyzed by real-time polymerase chain reaction. Protein phosphorylation of neutrophils was assessed by Western blot using phospho-specific antibodies. RESULTS: Stimulation with GM-CSF alone, but not tumor necrosis factor-alpha, was shown to increase the release of IL-1β and cleaved caspase-1 (p20) from human neutrophils. Tofacitinib, which inhibits GM-CSF–induced Janus kinase 2 (Jak2)-mediated signal transduction, completely abrogated GM-CSF–induced IL-1β and caspase-1 (p20) secretion from neutrophils. GM-CSF stimulation also induced pro-IL-1β mRNA expression in neutrophils and induced NLR family pyrin domain-containing 3 (NLRP3) protein expression. Although tofacitinib pretreatment marginally inhibited GM-CSF–induced pro-IL-1β mRNA expression, tofacitinib completely abrogated NLRP3 protein expression in neutrophils. CONCLUSIONS: These results indicate that GM-CSF signaling induces NLRP3 expression and subsequent IL-1β production by affecting neutrophils, which may cause the activation of innate immunity. Therefore, GM-CSF is a key regulator of the NLRP3 inflammasome and IL-1β production by activating innate immune cells. This process can be blocked by tofacitinib, which interferes with JAK/STAT signaling pathways. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13075-018-1685-x) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-6116484 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-61164842018-10-02 Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils Furuya, Makiko Yashiro Asano, Tomoyuki Sumichika, Yuya Sato, Shuzo Kobayashi, Hiroko Watanabe, Hiroshi Suzuki, Eiji Kozuru, Hideko Yatsuhashi, Hiroshi Koga, Tomohiro Ohira, Hiromasa Sekine, Hideharu Kawakami, Atsushi Migita, Kiyoshi Arthritis Res Ther Research Article BACKGROUND: Granulocyte–macrophage colony-stimulating factor (GM-CSF) has emerged as a crucial cytokine that activates myeloid cells to initiate tissue inflammation. However, the molecular actions of GM-CSF against innate immunity are still poorly characterized. Here, we investigated the in vitro effects of GM-CSF on the activation of human myeloid lineages, neutrophils, and the underlying intracellular signaling mechanism, including inflammasome activation. METHODS: Human neutrophils were stimulated with GM-CSF in the presence or absence of tofacitinib. The cellular supernatants were analyzed for interleukin-1 beta (IL-1β) and caspase-1 by enzyme-linked immunosorbent assay (ELISA) methods. Pro-IL-1β mRNA expressions in human neutrophils were analyzed by real-time polymerase chain reaction. Protein phosphorylation of neutrophils was assessed by Western blot using phospho-specific antibodies. RESULTS: Stimulation with GM-CSF alone, but not tumor necrosis factor-alpha, was shown to increase the release of IL-1β and cleaved caspase-1 (p20) from human neutrophils. Tofacitinib, which inhibits GM-CSF–induced Janus kinase 2 (Jak2)-mediated signal transduction, completely abrogated GM-CSF–induced IL-1β and caspase-1 (p20) secretion from neutrophils. GM-CSF stimulation also induced pro-IL-1β mRNA expression in neutrophils and induced NLR family pyrin domain-containing 3 (NLRP3) protein expression. Although tofacitinib pretreatment marginally inhibited GM-CSF–induced pro-IL-1β mRNA expression, tofacitinib completely abrogated NLRP3 protein expression in neutrophils. CONCLUSIONS: These results indicate that GM-CSF signaling induces NLRP3 expression and subsequent IL-1β production by affecting neutrophils, which may cause the activation of innate immunity. Therefore, GM-CSF is a key regulator of the NLRP3 inflammasome and IL-1β production by activating innate immune cells. This process can be blocked by tofacitinib, which interferes with JAK/STAT signaling pathways. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13075-018-1685-x) contains supplementary material, which is available to authorized users. BioMed Central 2018-08-29 2018 /pmc/articles/PMC6116484/ /pubmed/30157949 http://dx.doi.org/10.1186/s13075-018-1685-x Text en © The Author(s). 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Furuya, Makiko Yashiro Asano, Tomoyuki Sumichika, Yuya Sato, Shuzo Kobayashi, Hiroko Watanabe, Hiroshi Suzuki, Eiji Kozuru, Hideko Yatsuhashi, Hiroshi Koga, Tomohiro Ohira, Hiromasa Sekine, Hideharu Kawakami, Atsushi Migita, Kiyoshi Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils |
title | Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils |
title_full | Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils |
title_fullStr | Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils |
title_full_unstemmed | Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils |
title_short | Tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced NLRP3 inflammasome activation in human neutrophils |
title_sort | tofacitinib inhibits granulocyte–macrophage colony-stimulating factor-induced nlrp3 inflammasome activation in human neutrophils |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6116484/ https://www.ncbi.nlm.nih.gov/pubmed/30157949 http://dx.doi.org/10.1186/s13075-018-1685-x |
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