Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i>
<i>Mycobacterium tuberculosis</i> (<i>Mtb</i>), the causative agent of tuberculosis, secretes extracellular vesicles (EVs), which may play an important role in mediating interactions between bacteria and host cells. <i>Mtb</i> EVs can be isolated by means of vario...
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MDPI AG
2024-10-01
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| Online Access: | https://www.mdpi.com/2076-2607/12/11/2129 |
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| author | Wenjing Wang Yue Hou Jingfang Zhang Zhaogang Sun Hong Sun |
| author_facet | Wenjing Wang Yue Hou Jingfang Zhang Zhaogang Sun Hong Sun |
| author_sort | Wenjing Wang |
| collection | DOAJ |
| description | <i>Mycobacterium tuberculosis</i> (<i>Mtb</i>), the causative agent of tuberculosis, secretes extracellular vesicles (EVs), which may play an important role in mediating interactions between bacteria and host cells. <i>Mtb</i> EVs can be isolated by means of various techniques, which differ in terms of their effectiveness. In the present study, we found that an exosome isolation kit (EI) yielded higher numbers of EVs than either differential centrifugation (DC) or exosome detection via an ultrafast-isolation system (EXODUS). We also found that the EXODUS method revealed a greater abundance of H37Rv components within EVs, compared with the DC and EI methods. Analysis of the downstream application of H37Rv EVs revealed their internalization by RAW264.7 macrophages, peaking at 6 h, with subsequent activation of the TLR2 signaling pathway leading to the expression of inflammatory cytokines including IL-6 and TNF-α. It was also found that H37Rv EVs could cross the blood–brain barrier (BBB) and enter the brain, peaking at 12 h post-injection, eliciting an inflammatory response in the cerebral parenchyma, cerebellum, and hippocampus that persisted for up to 6 days. These findings offer novel insights into the pathogenesis of <i>Mtb</i>-induced diseases and may guide the development of therapeutic strategies. |
| format | Article |
| id | doaj-art-8d1bbe0568ac49978a84d2c8f3b689bd |
| institution | DOAJ |
| issn | 2076-2607 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj-art-8d1bbe0568ac49978a84d2c8f3b689bd2025-08-20T02:48:06ZengMDPI AGMicroorganisms2076-26072024-10-011211212910.3390/microorganisms12112129Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i>Wenjing Wang0Yue Hou1Jingfang Zhang2Zhaogang Sun3Hong Sun4Beijing Chest Hospital affiliated to Capital Medical University, Beijing 100000, ChinaBeijing Chest Hospital affiliated to Capital Medical University, Beijing 100000, ChinaBeijing Chest Hospital affiliated to Capital Medical University, Beijing 100000, ChinaBeijing Chest Hospital affiliated to Capital Medical University, Beijing 100000, ChinaBeijing Chest Hospital affiliated to Capital Medical University, Beijing 100000, China<i>Mycobacterium tuberculosis</i> (<i>Mtb</i>), the causative agent of tuberculosis, secretes extracellular vesicles (EVs), which may play an important role in mediating interactions between bacteria and host cells. <i>Mtb</i> EVs can be isolated by means of various techniques, which differ in terms of their effectiveness. In the present study, we found that an exosome isolation kit (EI) yielded higher numbers of EVs than either differential centrifugation (DC) or exosome detection via an ultrafast-isolation system (EXODUS). We also found that the EXODUS method revealed a greater abundance of H37Rv components within EVs, compared with the DC and EI methods. Analysis of the downstream application of H37Rv EVs revealed their internalization by RAW264.7 macrophages, peaking at 6 h, with subsequent activation of the TLR2 signaling pathway leading to the expression of inflammatory cytokines including IL-6 and TNF-α. It was also found that H37Rv EVs could cross the blood–brain barrier (BBB) and enter the brain, peaking at 12 h post-injection, eliciting an inflammatory response in the cerebral parenchyma, cerebellum, and hippocampus that persisted for up to 6 days. These findings offer novel insights into the pathogenesis of <i>Mtb</i>-induced diseases and may guide the development of therapeutic strategies.https://www.mdpi.com/2076-2607/12/11/2129mycobacteriaexosome detection via an ultrafast-isolation systemfunctional studyblood–brain barrier |
| spellingShingle | Wenjing Wang Yue Hou Jingfang Zhang Zhaogang Sun Hong Sun Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i> Microorganisms mycobacteria exosome detection via an ultrafast-isolation system functional study blood–brain barrier |
| title | Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i> |
| title_full | Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i> |
| title_fullStr | Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i> |
| title_full_unstemmed | Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i> |
| title_short | Improved Isolation Optimizes Downstream Application of Extracellular Vesicles Derived from <i>Mycobacterium tuberculosis</i> |
| title_sort | improved isolation optimizes downstream application of extracellular vesicles derived from i mycobacterium tuberculosis i |
| topic | mycobacteria exosome detection via an ultrafast-isolation system functional study blood–brain barrier |
| url | https://www.mdpi.com/2076-2607/12/11/2129 |
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