Serum extracellular vesicles profiling is associated with COVID‐19 progression and immune responses
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Abstract
Coronavirus disease 2019 (COVID‐19) has transformed very quickly into a world pandemic with severe and unexpected consequences on human health. Concerted efforts to generate better diagnostic and prognostic tools have been ongoing. Research, thus far, has primarily focused on the virus itself or the direct immune response to it. Here, we propose extracellular vesicles (EVs) from serum liquid biopsies as a new and unique modality to unify diagnostic and prognostic tools for COVID‐19 analyses. EVs are a novel player in intercellular signalling particularly influencing immune responses. We herein show that innate and adaptive immune EVs profiling, together with SARS‐CoV‐2 Spike S1 + EVs provide a novel signature for SARS‐CoV‐2 infection. It also provides a unique ability to associate the co‐existence of viral and host cell signatures to monitor affected tissues and severity of the disease progression. And provide a phenotypic insight into COVID‐associated EVs.
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SciScore for 10.1101/2022.01.19.22269529: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Consent: All participants, patients and healthy controls, signed a written informed consent.
IRB: This non-interventional, observational study was approved by the Cantonal Ethics Committee of Zurich (BASEC #2016-01440) and performed in accordance with the Declaration of Helsinki.Sex as a biological variable not detected. Randomization not detected. Blinding Investigators were blinded to disease severity, while performing experiments. Power Analysis The sample size was based on availability of the samples. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources All healthy controls were tested for SARS-CoV-2 specific IgA and IgG antibodies, and all were below the … SciScore for 10.1101/2022.01.19.22269529: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics Consent: All participants, patients and healthy controls, signed a written informed consent.
IRB: This non-interventional, observational study was approved by the Cantonal Ethics Committee of Zurich (BASEC #2016-01440) and performed in accordance with the Declaration of Helsinki.Sex as a biological variable not detected. Randomization not detected. Blinding Investigators were blinded to disease severity, while performing experiments. Power Analysis The sample size was based on availability of the samples. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources All healthy controls were tested for SARS-CoV-2 specific IgA and IgG antibodies, and all were below the diagnostic reference value. SARS-CoV-2 specific IgA and IgGsuggested: NoneTo test the antibody binding specificity, prior to immunostaining of Spike S1 on cells and EVs, molar ratios of 0:1, 1:1, 2:1, 5:1 of recombinant Spike S1 proteins (Sino Biological, #40591-V08H) were incubated with anti-Spike S1 for 30 mins at 4°C to neutralize unbound antibodies. anti-Spikesuggested: NoneExperimental Models: Cell Lines Sentences Resources Spike S1 signals were detected by flow cytometry (HEK293A) and nano flow analyzer (EVs). HEK293Asuggested: RRID:CVCL_6910)Recombinant DNA Sentences Resources To generate Spike S1 expressing HEK293A cells and EVs, we transfected HEK293A cells with pCMV14-3X-Flag-SARS-CoV-2 S plasmid (Addgene #145780) together with GFP plasmid (for transfection efficiency quantification) and EVs were collected 24 hours post transfection from conditioned medium using serial centrifugation as noted above. pCMV14-3X-Flag-SARS-CoV-2 Ssuggested: RRID:Addgene_145780)GFPsuggested: RRID:Addgene_126657)Software and Algorithms Sentences Resources Isolation of serum EVs for phenotyping analysis and functional assays: 1 mL of serum samples were first diluted with 9 mL of PBS and concentrated using Amicon® ultra-0.5 centrifugal filter devices (Millipore, Amicon® Ultra 100 K device) at 3,000 g for 30 mins at 4°C. Amicon®suggested: NoneData and statistical analysis: Both cells and EVs flow cytometry data were exported as FCS files and analyzed using Flowjo software. Flowjosuggested: (FlowJo, RRID:SCR_008520)Statistical analysis of clinical diagnostic and flow cytometry values were performed using Graphpad (version 9.1.1, GraphPad Software, La Jolla California USA) Graphpadsuggested: (GraphPad, RRID:SCR_000306)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.
Results from scite Reference Check: We found no unreliable references.
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