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Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses
INTRODUCTION: Exosomes are nanograde membrane-bound vesicles secreted from most cell types through the fusion of multivesicular bodies with plasma membranes. Some of these exosomes are well defined, and are known to have immunomodulatory properties as well as play critical roles in intercellular com...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove Medical Press
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5783023/ https://www.ncbi.nlm.nih.gov/pubmed/29403276 http://dx.doi.org/10.2147/IJN.S151110 |
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author | Li, Yawen Liu, Yuan Xiu, Fangming Wang, Jianing Cong, Hua He, Shenyi Shi, Yongyu Wang, Xiaoyan Li, Xun Zhou, Huaiyu |
author_facet | Li, Yawen Liu, Yuan Xiu, Fangming Wang, Jianing Cong, Hua He, Shenyi Shi, Yongyu Wang, Xiaoyan Li, Xun Zhou, Huaiyu |
author_sort | Li, Yawen |
collection | PubMed |
description | INTRODUCTION: Exosomes are nanograde membrane-bound vesicles secreted from most cell types through the fusion of multivesicular bodies with plasma membranes. Some of these exosomes are well defined, and are known to have immunomodulatory properties as well as play critical roles in intercellular communications. In this study, we characterized the exosomes derived from Toxoplasma gondii and their functions in aspect of immune responses. METHODS: T. gondii exosomes were isolated and identified using electron microscopy, nanoparticle tracking analysis, and Western blotting. The viability of macrophage RAW264.7 cells affected by exosomes was evaluated using a Cell Counting Kit (CCK-8). Then the uptake of T. gondii exosomes by RAW264.7 cells was detected by labeling with fluorescent dye PKH67. After exosomes stimulation, in vitro the production of interleukin (IL)-12, tumor necrosis factor (TNF)-α, interferon (IFN)-γ and IL-10 in RAW264.7 cells were investigated using enzyme-linked immunosorbent assay (ELISA). In immunized BALB/c mice, the antibodies, cytokines as well as the percentage of CD4+ and CD8+ T cells were determined using ELISA and flow cytometric analysis. Protective efficacy was evaluated by challenging intraperitoneally with tachyzoites of T. gondii. RESULTS: We successfully isolated and characterized the exosomes derived from T. gondii. Functionally, the viability of macrophage RAW264.7 cells was significantly affected by exosomes at a high concentration (160 μg/mL). The production of IL-12, TNF-α and IFN-γ in macrophage cells were increased, and the level of IL-10 was decreased. Furthermore, BALB/c mice immunized with T. gondii exosomes showed both humoral and cellular immune responses and also exhibited a prolonged survival time. CONCLUSION: T. gondii exosomes could modulate macrophage activation in vitro and trigger humoral and cellular immune responses and partial protection against acute parasite infection in mice, which suggested that exosomes may serve as a potential candidate against toxoplasmosis. |
format | Online Article Text |
id | pubmed-5783023 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Dove Medical Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-57830232018-02-05 Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses Li, Yawen Liu, Yuan Xiu, Fangming Wang, Jianing Cong, Hua He, Shenyi Shi, Yongyu Wang, Xiaoyan Li, Xun Zhou, Huaiyu Int J Nanomedicine Original Research INTRODUCTION: Exosomes are nanograde membrane-bound vesicles secreted from most cell types through the fusion of multivesicular bodies with plasma membranes. Some of these exosomes are well defined, and are known to have immunomodulatory properties as well as play critical roles in intercellular communications. In this study, we characterized the exosomes derived from Toxoplasma gondii and their functions in aspect of immune responses. METHODS: T. gondii exosomes were isolated and identified using electron microscopy, nanoparticle tracking analysis, and Western blotting. The viability of macrophage RAW264.7 cells affected by exosomes was evaluated using a Cell Counting Kit (CCK-8). Then the uptake of T. gondii exosomes by RAW264.7 cells was detected by labeling with fluorescent dye PKH67. After exosomes stimulation, in vitro the production of interleukin (IL)-12, tumor necrosis factor (TNF)-α, interferon (IFN)-γ and IL-10 in RAW264.7 cells were investigated using enzyme-linked immunosorbent assay (ELISA). In immunized BALB/c mice, the antibodies, cytokines as well as the percentage of CD4+ and CD8+ T cells were determined using ELISA and flow cytometric analysis. Protective efficacy was evaluated by challenging intraperitoneally with tachyzoites of T. gondii. RESULTS: We successfully isolated and characterized the exosomes derived from T. gondii. Functionally, the viability of macrophage RAW264.7 cells was significantly affected by exosomes at a high concentration (160 μg/mL). The production of IL-12, TNF-α and IFN-γ in macrophage cells were increased, and the level of IL-10 was decreased. Furthermore, BALB/c mice immunized with T. gondii exosomes showed both humoral and cellular immune responses and also exhibited a prolonged survival time. CONCLUSION: T. gondii exosomes could modulate macrophage activation in vitro and trigger humoral and cellular immune responses and partial protection against acute parasite infection in mice, which suggested that exosomes may serve as a potential candidate against toxoplasmosis. Dove Medical Press 2018-01-19 /pmc/articles/PMC5783023/ /pubmed/29403276 http://dx.doi.org/10.2147/IJN.S151110 Text en © 2018 Li et al. This work is published and licensed by Dove Medical Press Limited The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. |
spellingShingle | Original Research Li, Yawen Liu, Yuan Xiu, Fangming Wang, Jianing Cong, Hua He, Shenyi Shi, Yongyu Wang, Xiaoyan Li, Xun Zhou, Huaiyu Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses |
title | Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses |
title_full | Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses |
title_fullStr | Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses |
title_full_unstemmed | Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses |
title_short | Characterization of exosomes derived from Toxoplasma gondii and their functions in modulating immune responses |
title_sort | characterization of exosomes derived from toxoplasma gondii and their functions in modulating immune responses |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5783023/ https://www.ncbi.nlm.nih.gov/pubmed/29403276 http://dx.doi.org/10.2147/IJN.S151110 |
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