Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients
BackgroundThe global outbreak of COVID-19, and the limited availability of clinical treatments, forced researchers around the world to search for the pathogenesis and potential treatments. Understanding the pathogenesis of SARS-CoV-2 is crucial to respond better to the current coronavirus disease 20...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1052141/full |
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| author | Ruiting Sun Yanling Cai Yanling Cai Yumin Zhou Ge Bai Airu Zhu Panyue Kong Jing Sun Yimin Li Yuefei Liu Wenting Liao Jiye Liu Nan Cui Jinsheng Xiang Bing Li Jincun Zhao Di Wu Di Wu Pixin Ran |
| author_facet | Ruiting Sun Yanling Cai Yanling Cai Yumin Zhou Ge Bai Airu Zhu Panyue Kong Jing Sun Yimin Li Yuefei Liu Wenting Liao Jiye Liu Nan Cui Jinsheng Xiang Bing Li Jincun Zhao Di Wu Di Wu Pixin Ran |
| author_sort | Ruiting Sun |
| collection | DOAJ |
| description | BackgroundThe global outbreak of COVID-19, and the limited availability of clinical treatments, forced researchers around the world to search for the pathogenesis and potential treatments. Understanding the pathogenesis of SARS-CoV-2 is crucial to respond better to the current coronavirus disease 2019 (COVID-19) pandemic.MethodsWe collected sputum samples from 20 COVID-19 patients and healthy controls. Transmission electron microscopy was used to observe the morphology of SARS-CoV-2. Extracellular vesicles (EVs) were isolated from sputum and the supernatant of VeroE6 cells, and were characterized by transmission electron microscopy, nanoparticle tracking analysis and Western-Blotting. Furthermore, a proximity barcoding assay was used to investigate immune-related proteins in single EV, and the relationship between EVs and SARS-CoV-2.ResultTransmission electron microscopy images of SARS-COV-2 virus reveal EV-like vesicles around the virion, and western blot analysis of EVs extracted from the supernatant of SARS-COV-2-infected VeroE6 cells showed that they expressed SARS-COV-2 protein. These EVs have the infectivity of SARS-COV-2, and the addition can cause the infection and damage of normal VeroE6 cells. In addition, EVs derived from the sputum of patients infected with SARS-COV-2 expressed high levels of IL6 and TGF-β, which correlated strongly with expression of the SARS-CoV-2 N protein. Among 40 EV subpopulations identified, 18 differed significantly between patients and controls. The EV subpopulation regulated by CD81 was the most likely to correlate with changes in the pulmonary microenvironment after SARS-CoV-2 infection. Single extracellular vesicles in the sputum of COVID-19 patients harbor infection-mediated alterations in host and virus-derived proteins.ConclusionsThese results demonstrate that EVs derived from the sputum of patients participate in virus infection and immune responses. This study provides evidence of an association between EVs and SARS-CoV-2, providing insight into the possible pathogenesis of SARS-CoV-2 infection and the possibility of developing nanoparticle-based antiviral drugs. |
| first_indexed | 2024-04-09T13:07:45Z |
| format | Article |
| id | doaj.art-2d9b4e9939944a6187fed4ac040629bf |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-09T13:07:45Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-2d9b4e9939944a6187fed4ac040629bf2023-05-12T13:17:18ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-05-011410.3389/fimmu.2023.10521411052141Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patientsRuiting Sun0Yanling Cai1Yanling Cai2Yumin Zhou3Ge Bai4Airu Zhu5Panyue Kong6Jing Sun7Yimin Li8Yuefei Liu9Wenting Liao10Jiye Liu11Nan Cui12Jinsheng Xiang13Bing Li14Jincun Zhao15Di Wu16Di Wu17Pixin Ran18National Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaShenzhen Second People’s Hospital, Postdoctoral Workstation of Zhongshan School of Medicine, Sun Yat-Sen University, Shenzhen, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaR&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, ChinaR&D Department, Vesicode AB, Solna, SwedenNational Center for Respiratory Medicine, State Key Laboratory of Respiratory Disease, National Clinical Researcher Center for Respiratory Diseases, Guangzhou Institute of Respiratory Health, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, ChinaBackgroundThe global outbreak of COVID-19, and the limited availability of clinical treatments, forced researchers around the world to search for the pathogenesis and potential treatments. Understanding the pathogenesis of SARS-CoV-2 is crucial to respond better to the current coronavirus disease 2019 (COVID-19) pandemic.MethodsWe collected sputum samples from 20 COVID-19 patients and healthy controls. Transmission electron microscopy was used to observe the morphology of SARS-CoV-2. Extracellular vesicles (EVs) were isolated from sputum and the supernatant of VeroE6 cells, and were characterized by transmission electron microscopy, nanoparticle tracking analysis and Western-Blotting. Furthermore, a proximity barcoding assay was used to investigate immune-related proteins in single EV, and the relationship between EVs and SARS-CoV-2.ResultTransmission electron microscopy images of SARS-COV-2 virus reveal EV-like vesicles around the virion, and western blot analysis of EVs extracted from the supernatant of SARS-COV-2-infected VeroE6 cells showed that they expressed SARS-COV-2 protein. These EVs have the infectivity of SARS-COV-2, and the addition can cause the infection and damage of normal VeroE6 cells. In addition, EVs derived from the sputum of patients infected with SARS-COV-2 expressed high levels of IL6 and TGF-β, which correlated strongly with expression of the SARS-CoV-2 N protein. Among 40 EV subpopulations identified, 18 differed significantly between patients and controls. The EV subpopulation regulated by CD81 was the most likely to correlate with changes in the pulmonary microenvironment after SARS-CoV-2 infection. Single extracellular vesicles in the sputum of COVID-19 patients harbor infection-mediated alterations in host and virus-derived proteins.ConclusionsThese results demonstrate that EVs derived from the sputum of patients participate in virus infection and immune responses. This study provides evidence of an association between EVs and SARS-CoV-2, providing insight into the possible pathogenesis of SARS-CoV-2 infection and the possibility of developing nanoparticle-based antiviral drugs.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1052141/fullCOVID-19SARS-CoV-2single extracellular vesiclesinflammatory responseextracellular vesicles subpopulation |
| spellingShingle | Ruiting Sun Yanling Cai Yanling Cai Yumin Zhou Ge Bai Airu Zhu Panyue Kong Jing Sun Yimin Li Yuefei Liu Wenting Liao Jiye Liu Nan Cui Jinsheng Xiang Bing Li Jincun Zhao Di Wu Di Wu Pixin Ran Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients Frontiers in Immunology COVID-19 SARS-CoV-2 single extracellular vesicles inflammatory response extracellular vesicles subpopulation |
| title | Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients |
| title_full | Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients |
| title_fullStr | Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients |
| title_full_unstemmed | Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients |
| title_short | Proteomic profiling of single extracellular vesicles reveals colocalization of SARS-CoV-2 with a CD81/integrin-rich EV subpopulation in sputum from COVID-19 severe patients |
| title_sort | proteomic profiling of single extracellular vesicles reveals colocalization of sars cov 2 with a cd81 integrin rich ev subpopulation in sputum from covid 19 severe patients |
| topic | COVID-19 SARS-CoV-2 single extracellular vesicles inflammatory response extracellular vesicles subpopulation |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1052141/full |
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