Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli

A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation. Ausführliche Beschreibung

Gespeichert in:

Autor*in:	Praveen Akuthota [verfasserIn] Lívia A. S. Carmo [verfasserIn] Kennedy Bonjour [verfasserIn] Ryann Murphy [verfasserIn] Thiago P. Silva [verfasserIn] Juliana P. Gamalier [verfasserIn] Kelsey Capron [verfasserIn] John Tigges [verfasserIn] Vasilis Toxavidis [verfasserIn] Virginia Camacho [verfasserIn] Ionita Ghiran [verfasserIn] Shigeharu Ueki [verfasserIn] Peter F. Weller [verfasserIn] Rossana C.N. Melo [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2016

Schlagwörter:	Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α)

Übergeordnetes Werk:	In: Frontiers in Cell and Developmental Biology - Frontiers Media S.A., 2014, 4(2016)
Übergeordnetes Werk:	volume:4 ; year:2016

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.3389/fcell.2016.00117

Katalog-ID:	DOAJ00788401X

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allfields	10.3389/fcell.2016.00117 doi (DE-627)DOAJ00788401X (DE-599)DOAJ108a8a59009849d4aa1fcb79b635ecfc DE-627 ger DE-627 rakwb eng QH301-705.5 Praveen Akuthota verfasserin aut Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli 2016 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation. Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α) Biology (General) Praveen Akuthota verfasserin aut Lívia A. S. Carmo verfasserin aut Kennedy Bonjour verfasserin aut Ryann Murphy verfasserin aut Thiago P. Silva verfasserin aut Juliana P. Gamalier verfasserin aut Kelsey Capron verfasserin aut John Tigges verfasserin aut Vasilis Toxavidis verfasserin aut Virginia Camacho verfasserin aut Ionita Ghiran verfasserin aut Shigeharu Ueki verfasserin aut Peter F. Weller verfasserin aut Rossana C.N. Melo verfasserin aut Rossana C.N. Melo verfasserin aut In Frontiers in Cell and Developmental Biology Frontiers Media S.A., 2014 4(2016) (DE-627)770398138 (DE-600)2737824-X 2296634X nnns volume:4 year:2016 https://doi.org/10.3389/fcell.2016.00117 kostenfrei https://doaj.org/article/108a8a59009849d4aa1fcb79b635ecfc kostenfrei http://journal.frontiersin.org/Journal/10.3389/fcell.2016.00117/full kostenfrei https://doaj.org/toc/2296-634X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2016
spelling	10.3389/fcell.2016.00117 doi (DE-627)DOAJ00788401X (DE-599)DOAJ108a8a59009849d4aa1fcb79b635ecfc DE-627 ger DE-627 rakwb eng QH301-705.5 Praveen Akuthota verfasserin aut Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli 2016 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation. Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α) Biology (General) Praveen Akuthota verfasserin aut Lívia A. S. Carmo verfasserin aut Kennedy Bonjour verfasserin aut Ryann Murphy verfasserin aut Thiago P. Silva verfasserin aut Juliana P. Gamalier verfasserin aut Kelsey Capron verfasserin aut John Tigges verfasserin aut Vasilis Toxavidis verfasserin aut Virginia Camacho verfasserin aut Ionita Ghiran verfasserin aut Shigeharu Ueki verfasserin aut Peter F. Weller verfasserin aut Rossana C.N. Melo verfasserin aut Rossana C.N. Melo verfasserin aut In Frontiers in Cell and Developmental Biology Frontiers Media S.A., 2014 4(2016) (DE-627)770398138 (DE-600)2737824-X 2296634X nnns volume:4 year:2016 https://doi.org/10.3389/fcell.2016.00117 kostenfrei https://doaj.org/article/108a8a59009849d4aa1fcb79b635ecfc kostenfrei http://journal.frontiersin.org/Journal/10.3389/fcell.2016.00117/full kostenfrei https://doaj.org/toc/2296-634X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2016
allfields_unstemmed	10.3389/fcell.2016.00117 doi (DE-627)DOAJ00788401X (DE-599)DOAJ108a8a59009849d4aa1fcb79b635ecfc DE-627 ger DE-627 rakwb eng QH301-705.5 Praveen Akuthota verfasserin aut Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli 2016 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation. Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α) Biology (General) Praveen Akuthota verfasserin aut Lívia A. S. Carmo verfasserin aut Kennedy Bonjour verfasserin aut Ryann Murphy verfasserin aut Thiago P. Silva verfasserin aut Juliana P. Gamalier verfasserin aut Kelsey Capron verfasserin aut John Tigges verfasserin aut Vasilis Toxavidis verfasserin aut Virginia Camacho verfasserin aut Ionita Ghiran verfasserin aut Shigeharu Ueki verfasserin aut Peter F. Weller verfasserin aut Rossana C.N. Melo verfasserin aut Rossana C.N. Melo verfasserin aut In Frontiers in Cell and Developmental Biology Frontiers Media S.A., 2014 4(2016) (DE-627)770398138 (DE-600)2737824-X 2296634X nnns volume:4 year:2016 https://doi.org/10.3389/fcell.2016.00117 kostenfrei https://doaj.org/article/108a8a59009849d4aa1fcb79b635ecfc kostenfrei http://journal.frontiersin.org/Journal/10.3389/fcell.2016.00117/full kostenfrei https://doaj.org/toc/2296-634X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2016
allfieldsGer	10.3389/fcell.2016.00117 doi (DE-627)DOAJ00788401X (DE-599)DOAJ108a8a59009849d4aa1fcb79b635ecfc DE-627 ger DE-627 rakwb eng QH301-705.5 Praveen Akuthota verfasserin aut Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli 2016 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation. Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α) Biology (General) Praveen Akuthota verfasserin aut Lívia A. S. Carmo verfasserin aut Kennedy Bonjour verfasserin aut Ryann Murphy verfasserin aut Thiago P. Silva verfasserin aut Juliana P. Gamalier verfasserin aut Kelsey Capron verfasserin aut John Tigges verfasserin aut Vasilis Toxavidis verfasserin aut Virginia Camacho verfasserin aut Ionita Ghiran verfasserin aut Shigeharu Ueki verfasserin aut Peter F. Weller verfasserin aut Rossana C.N. Melo verfasserin aut Rossana C.N. Melo verfasserin aut In Frontiers in Cell and Developmental Biology Frontiers Media S.A., 2014 4(2016) (DE-627)770398138 (DE-600)2737824-X 2296634X nnns volume:4 year:2016 https://doi.org/10.3389/fcell.2016.00117 kostenfrei https://doaj.org/article/108a8a59009849d4aa1fcb79b635ecfc kostenfrei http://journal.frontiersin.org/Journal/10.3389/fcell.2016.00117/full kostenfrei https://doaj.org/toc/2296-634X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2016
allfieldsSound	10.3389/fcell.2016.00117 doi (DE-627)DOAJ00788401X (DE-599)DOAJ108a8a59009849d4aa1fcb79b635ecfc DE-627 ger DE-627 rakwb eng QH301-705.5 Praveen Akuthota verfasserin aut Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli 2016 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation. Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α) Biology (General) Praveen Akuthota verfasserin aut Lívia A. S. Carmo verfasserin aut Kennedy Bonjour verfasserin aut Ryann Murphy verfasserin aut Thiago P. Silva verfasserin aut Juliana P. Gamalier verfasserin aut Kelsey Capron verfasserin aut John Tigges verfasserin aut Vasilis Toxavidis verfasserin aut Virginia Camacho verfasserin aut Ionita Ghiran verfasserin aut Shigeharu Ueki verfasserin aut Peter F. Weller verfasserin aut Rossana C.N. Melo verfasserin aut Rossana C.N. Melo verfasserin aut In Frontiers in Cell and Developmental Biology Frontiers Media S.A., 2014 4(2016) (DE-627)770398138 (DE-600)2737824-X 2296634X nnns volume:4 year:2016 https://doi.org/10.3389/fcell.2016.00117 kostenfrei https://doaj.org/article/108a8a59009849d4aa1fcb79b635ecfc kostenfrei http://journal.frontiersin.org/Journal/10.3389/fcell.2016.00117/full kostenfrei https://doaj.org/toc/2296-634X Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 4 2016
language	English
source	In Frontiers in Cell and Developmental Biology 4(2016) volume:4 year:2016
sourceStr	In Frontiers in Cell and Developmental Biology 4(2016) volume:4 year:2016
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α) Biology (General)
isfreeaccess_bool	true
container_title	Frontiers in Cell and Developmental Biology
authorswithroles_txt_mv	Praveen Akuthota @@aut@@ Lívia A. S. Carmo @@aut@@ Kennedy Bonjour @@aut@@ Ryann Murphy @@aut@@ Thiago P. Silva @@aut@@ Juliana P. Gamalier @@aut@@ Kelsey Capron @@aut@@ John Tigges @@aut@@ Vasilis Toxavidis @@aut@@ Virginia Camacho @@aut@@ Ionita Ghiran @@aut@@ Shigeharu Ueki @@aut@@ Peter F. Weller @@aut@@ Rossana C.N. Melo @@aut@@
publishDateDaySort_date	2016-01-01T00:00:00Z
hierarchy_top_id	770398138
id	DOAJ00788401X
language_de	englisch
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callnumber-first	Q - Science
author	Praveen Akuthota
spellingShingle	Praveen Akuthota misc QH301-705.5 misc Inflammation misc Tetraspanins misc CD9 misc CD63 misc Transmission electron microscopy misc Tumor necrosis factor alpha (TNF-α) misc Biology (General) Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli
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topic_title	QH301-705.5 Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli Inflammation Tetraspanins CD9 CD63 Transmission electron microscopy Tumor necrosis factor alpha (TNF-α)
topic	misc QH301-705.5 misc Inflammation misc Tetraspanins misc CD9 misc CD63 misc Transmission electron microscopy misc Tumor necrosis factor alpha (TNF-α) misc Biology (General)
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title	Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli
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title_full	Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli
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author_browse	Praveen Akuthota Lívia A. S. Carmo Kennedy Bonjour Ryann Murphy Thiago P. Silva Juliana P. Gamalier Kelsey Capron John Tigges Vasilis Toxavidis Virginia Camacho Ionita Ghiran Shigeharu Ueki Peter F. Weller Rossana C.N. Melo
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title_sort	extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli
callnumber	QH301-705.5
title_auth	Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli
abstract	A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation.
abstractGer	A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation.
abstract_unstemmed	A key function of human eosinophils is to secrete cytokines, chemokines and cationic proteins, trafficking and releasing these mediators for roles in inflammation and other immune responses. Eosinophil activation leads to secretion of pre-synthesized granule-stored mediators through different mechanisms, but the ability of eosinophils to secrete extracellular vesicles (EVs), very small vesicles with preserved membrane topology, is still poorly understood. In the present work, we sought to identify and characterize EVs released from human eosinophils during different conditions: after a culturing period or after isolation and stimulation with inflammatory stimuli, which are known to induce eosinophil activation and secretion: CCL11 (eotaxin-1) and tumor necrosis factor alpha (TNF-α). EV production was investigated by nanoscale flow cytometry, conventional transmission electron microscopy (TEM) and pre-embedding immunonanogold EM. The tetraspanins CD63 and CD9 were used as EV biomarkers for both flow cytometry and ultrastructural immunolabeling. Nanoscale flow cytometry showed that human eosinophils produce EVs in culture and that a population of EVs expressed detectable CD9, while CD63 was not consistently detected. When eosinophils were stimulated immediately after isolation and analyzed by TEM, EVs were clearly identified as microvesicles (MVs)outwardly budding off the plasma membrane. Both CCL11 and TNF-α induced significant increases of MVs compared to unstimulated cells.TNF-α induced amplified release of MVs more than CCL11. Eosinophil MV diameters varied from 20-1000 nm. Immunonanogold EM revealed clear immunolabeling for CD63 and CD9 on eosinophil MVs, although not all MVs were labeled. Altogether, we identified, for the first time, that human eosinophils secrete MVs and that this production increases in response to inflammatory stimuli. This is important to understand the complex secretory activities of eosinophils underlying immune responses. The contribution of the eosinophil-derived MVs to the regulation of immune responses awaits further investigation.
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title_short	Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli
url	https://doi.org/10.3389/fcell.2016.00117 https://doaj.org/article/108a8a59009849d4aa1fcb79b635ecfc http://journal.frontiersin.org/Journal/10.3389/fcell.2016.00117/full https://doaj.org/toc/2296-634X
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Extracellular microvesicle production by human eosinophils activated by “inflammatory” stimuli

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