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

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

doi:10.3389/fimmu.2023.1052141

Frontiers in Immunology (May 2023)

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

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

Affiliations

Ruiting Sun: National 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, China
Yanling Cai: Shenzhen Second People’s Hospital, Postdoctoral Workstation of Zhongshan School of Medicine, Sun Yat-Sen University, Shenzhen, Guangdong, China
Yanling Cai: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Yumin Zhou: National 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, China
Ge Bai: National 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, China
Airu Zhu: National 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, China
Panyue Kong: National 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, China
Jing Sun: National 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, China
Yimin Li: National 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, China
Yuefei Liu: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Wenting Liao: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Jiye Liu: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Nan Cui: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Jinsheng Xiang: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Bing Li: National 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, China
Jincun Zhao: National 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, China
Di Wu: R&D Department, Shenzhen Secretech Co. Ltd, Shenzhen, Guangdong, China
Di Wu: R&D Department, Vesicode AB, Solna, Sweden
Pixin Ran: National 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, China

DOI: https://doi.org/10.3389/fimmu.2023.1052141
Journal volume & issue: Vol. 14

Abstract

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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.

Published in Frontiers in Immunology

ISSN: 1664-3224 (Online)
Publisher: Frontiers Media S.A.
Country of publisher: Switzerland
LCC subjects: Medicine: Internal medicine: Specialties of internal medicine: Immunologic diseases. Allergy
Website: http://journal.frontiersin.org/journal/immunology

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