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Inflammation leads to distinct populations of extracellular vesicles from microglia

BACKGROUND: Activated microglia play an essential role in inflammatory responses elicited in the central nervous system (CNS). Microglia-derived extracellular vesicles (EVs) are suggested to be involved in propagation of inflammatory signals and in the modulation of cell-to-cell communication. Howev...

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Autores principales: Yang, Yiyi, Boza-Serrano, Antonio, Dunning, Christopher J. R., Clausen, Bettina Hjelm, Lambertsen, Kate Lykke, Deierborg, Tomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5972400/
https://www.ncbi.nlm.nih.gov/pubmed/29807527
http://dx.doi.org/10.1186/s12974-018-1204-7
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author Yang, Yiyi
Boza-Serrano, Antonio
Dunning, Christopher J. R.
Clausen, Bettina Hjelm
Lambertsen, Kate Lykke
Deierborg, Tomas
author_facet Yang, Yiyi
Boza-Serrano, Antonio
Dunning, Christopher J. R.
Clausen, Bettina Hjelm
Lambertsen, Kate Lykke
Deierborg, Tomas
author_sort Yang, Yiyi
collection PubMed
description BACKGROUND: Activated microglia play an essential role in inflammatory responses elicited in the central nervous system (CNS). Microglia-derived extracellular vesicles (EVs) are suggested to be involved in propagation of inflammatory signals and in the modulation of cell-to-cell communication. However, there is a lack of knowledge on the regulation of EVs and how this in turn facilitates the communication between cells in the brain. Here, we characterized microglial EVs under inflammatory conditions and investigated the effects of inflammation on the EV size, quantity, and protein content. METHODS: We have utilized western blot, nanoparticle tracking analysis (NTA), and mass spectrometry to characterize EVs and examine the alterations of secreted EVs from a microglial cell line (BV2) following lipopolysaccharide (LPS) and tumor necrosis factor (TNF) inhibitor (etanercept) treatments, or either alone. The inflammatory responses were measured with multiplex cytokine ELISA and western blot. We also subjected TNF knockout mice to experimental stroke (permanent middle cerebral artery occlusion) and validated the effect of TNF inhibition on EV release. RESULTS: Our analysis of EVs originating from activated BV2 microglia revealed a significant increase in the intravesicular levels of TNF and interleukin (IL)-6. We also observed that the number of EVs released was reduced both in vitro and in vivo when inflammation was inhibited via the TNF pathway. Finally, via mass spectrometry, we identified 49 unique proteins in EVs released from LPS-activated microglia compared to control EVs (58 proteins in EVs released from LPS-activated microglia and 37 from control EVs). According to Gene Ontology (GO) analysis, we found a large increase of proteins related to translation and transcription in EVs from LPS. Importantly, we showed a distinct profile of proteins found in EVs released from LPS treated cells compared to control. CONCLUSIONS: We demonstrate altered EV production in BV2 microglial cells and altered cytokine levels and protein composition carried by EVs in response to LPS challenge. Our findings provide new insights into the potential roles of EVs that could be related to the pathogenesis in neuroinflammatory diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12974-018-1204-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-59724002018-06-05 Inflammation leads to distinct populations of extracellular vesicles from microglia Yang, Yiyi Boza-Serrano, Antonio Dunning, Christopher J. R. Clausen, Bettina Hjelm Lambertsen, Kate Lykke Deierborg, Tomas J Neuroinflammation Research BACKGROUND: Activated microglia play an essential role in inflammatory responses elicited in the central nervous system (CNS). Microglia-derived extracellular vesicles (EVs) are suggested to be involved in propagation of inflammatory signals and in the modulation of cell-to-cell communication. However, there is a lack of knowledge on the regulation of EVs and how this in turn facilitates the communication between cells in the brain. Here, we characterized microglial EVs under inflammatory conditions and investigated the effects of inflammation on the EV size, quantity, and protein content. METHODS: We have utilized western blot, nanoparticle tracking analysis (NTA), and mass spectrometry to characterize EVs and examine the alterations of secreted EVs from a microglial cell line (BV2) following lipopolysaccharide (LPS) and tumor necrosis factor (TNF) inhibitor (etanercept) treatments, or either alone. The inflammatory responses were measured with multiplex cytokine ELISA and western blot. We also subjected TNF knockout mice to experimental stroke (permanent middle cerebral artery occlusion) and validated the effect of TNF inhibition on EV release. RESULTS: Our analysis of EVs originating from activated BV2 microglia revealed a significant increase in the intravesicular levels of TNF and interleukin (IL)-6. We also observed that the number of EVs released was reduced both in vitro and in vivo when inflammation was inhibited via the TNF pathway. Finally, via mass spectrometry, we identified 49 unique proteins in EVs released from LPS-activated microglia compared to control EVs (58 proteins in EVs released from LPS-activated microglia and 37 from control EVs). According to Gene Ontology (GO) analysis, we found a large increase of proteins related to translation and transcription in EVs from LPS. Importantly, we showed a distinct profile of proteins found in EVs released from LPS treated cells compared to control. CONCLUSIONS: We demonstrate altered EV production in BV2 microglial cells and altered cytokine levels and protein composition carried by EVs in response to LPS challenge. Our findings provide new insights into the potential roles of EVs that could be related to the pathogenesis in neuroinflammatory diseases. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s12974-018-1204-7) contains supplementary material, which is available to authorized users. BioMed Central 2018-05-28 /pmc/articles/PMC5972400/ /pubmed/29807527 http://dx.doi.org/10.1186/s12974-018-1204-7 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
Yang, Yiyi
Boza-Serrano, Antonio
Dunning, Christopher J. R.
Clausen, Bettina Hjelm
Lambertsen, Kate Lykke
Deierborg, Tomas
Inflammation leads to distinct populations of extracellular vesicles from microglia
title Inflammation leads to distinct populations of extracellular vesicles from microglia
title_full Inflammation leads to distinct populations of extracellular vesicles from microglia
title_fullStr Inflammation leads to distinct populations of extracellular vesicles from microglia
title_full_unstemmed Inflammation leads to distinct populations of extracellular vesicles from microglia
title_short Inflammation leads to distinct populations of extracellular vesicles from microglia
title_sort inflammation leads to distinct populations of extracellular vesicles from microglia
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5972400/
https://www.ncbi.nlm.nih.gov/pubmed/29807527
http://dx.doi.org/10.1186/s12974-018-1204-7
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