Exosomes from COVID-19 Patients Carry Tenascin-C and Fibrinogen-β in Triggering Inflammatory Signals in Cells of Distant Organ
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Abstract
SARS-CoV-2 infection can cause cytokine storm and may overshoot immunity in humans; however, it remains to be determined whether virus-induced soluble mediators from infected cells are carried by exosomes as vehicles to distant organs and cause tissue damage in COVID-19 patients. We took an unbiased proteomic approach for analyses of exosomes isolated from plasma of healthy volunteers and COVID-19 patients. Our results revealed that tenascin-C (TNC) and fibrinogen-β (FGB) are highly abundant in exosomes from COVID-19 patients’ plasma compared with that of healthy normal controls. Since TNC and FGB stimulate pro-inflammatory cytokines via the Nuclear factor-κB (NF-κB) pathway, we examined the status of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and C–C motif chemokine ligand 5 (CCL5) expression upon exposure of hepatocytes to exosomes from COVID-19 patients and observed significant increase compared with that from healthy subjects. Together, our results demonstrate that TNC and FGB are transported through plasma exosomes and potentially trigger pro-inflammatory cytokine signaling in cells of distant organ.
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SciScore for 10.1101/2021.02.08.430369: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: This study was waived by the Saint Louis University Institutional Review Board for use of de-identified clinical specimens Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Western blot analysis: Cells lysates were subjected to Western blot analysis using specific antibodies to CD63 (Santa Cruz Biotechnology), TSG101 (Santa Cruz Biotechnology), tenascin (TNC) (Sigma), fibrinogen-β CD63suggested: NoneTSG101suggested: Nonetenascin ( TNCsuggested: NoneThe blot was reprobed with actin-HRP antibody (Santa Cruz … SciScore for 10.1101/2021.02.08.430369: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: This study was waived by the Saint Louis University Institutional Review Board for use of de-identified clinical specimens Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Western blot analysis: Cells lysates were subjected to Western blot analysis using specific antibodies to CD63 (Santa Cruz Biotechnology), TSG101 (Santa Cruz Biotechnology), tenascin (TNC) (Sigma), fibrinogen-β CD63suggested: NoneTSG101suggested: Nonetenascin ( TNCsuggested: NoneThe blot was reprobed with actin-HRP antibody (Santa Cruz Biotechnology) to compare protein load in each lane. actin-HRPsuggested: NoneExperimental Models: Cell Lines Sentences Resources Cell culture and exposure with exosomes: Immortalized human hepatocytes (IHH) and a human hepatoma cell line (Huh7) were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin at 37°C in a 5% CO2 atmosphere. Huh7suggested: NoneSoftware and Algorithms Sentences Resources Densitometry analysis was done using Image J software. Image Jsuggested: (ImageJ, RRID:SCR_003070)Pearson’s correlation analysis was performed using GraphPad Prism software. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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