Probiotic consortia improve anti-viral immunity to SARS-CoV-2 in Ferrets

This article has been Reviewed by the following groups

Read the full article

Abstract

Probiotics have been suggested as one solution to counter detrimental health effects by SARS-CoV-2, however, data so far is scarce. We tested the effect of two probiotic consortia, OL-1 and OL-2, against SARS-CoV-2 in ferrets and assessed their effect on cytokine production and transcriptome in a human monocyte-derived macrophage (Mf) and dendritic cell (DC) model.

The results showed that the consortia significantly reduced the viral load, modulated immune response, and regulated viral receptor expression in ferrets compared to placebo. In human Mf and DC model, OL-1 and OL-2 induced cytokine production and genes related to SARS-CoV-2 anti-viral immunity.

The study results indicate that probiotic stimulation of the ferret immune system leads to improved anti-viral immunity against SARS-COV-2 and that critical genes and cytokines for anti-SARS-CoV-2 immunity are stimulated in human immune cells in vitro . The effect of the consortia against SARS-CoV-2 warrants further investigations in human clinical trials.

Article activity feed

  1. SciScore for 10.1101/2021.07.23.453521: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    EthicsIACUC: The study was performed under American Association for Laboratory Animal Science (IACUC) protocol #20-031 RT-qPCR SARS-CoV-2 Titers: The concentration of virus in nasal washes from study days D1, D3, D5, D7, D9, and D10 was determined by RT-qPCR assay.
    IRB: The use of human blood was approved by the Ethics Committee of the Hospital District of Helsinki and Uusimaa, Finland (216/13/03/00/2016)
    Sex as a biological variableAll the strains are owned by and were manufactured along with placebo by IFF (Madison, Wisconsin, USA) Ferret studies: Outbred male ferrets
    RandomizationFerrets were randomly assigned to the different experimental groups.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    To detect the ACE2 by immunohistochemistry, Goat Anti-ACE2 antibody (R&D, Cat#: AF933, Lot#: HOK0620051) was used as the primary antibody.
    Anti-ACE2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Bifidobacterium animalis subsp. lactis Bl-04, American Type Culture Collection (ATCC) Safe Deposit (SD)5219, DGCC2908, Lacticaseibacillus paracasei subsp. paracasei Lpc-37, SD5275, ATCC PTA-4798, Lacticaseibacillus rhamnosus Lr-32, SD5217, and Ligilactobacillus salivarius Ls-33, SD5208, and for OL-2: Bifidobacterium animalis subsp. lactis Bi-07, SD5220, DGCC 2907, Lactobacillus acidophilus NCFM, SD5221, ATCC 700396, Limosilactobacillus fermentum SBS-1, DGCC1925, Lactococcus lactis subsp. lactis Ll-23, DGCC8656, Streptococcus thermophilus subsp. thermophilus St-21, SD5207.
    SD5275
    suggested: None
    Recombinant DNA
    SentencesResources
    PBMCs without stimulation were used as a control, dimethyl sulfoxide (DMSO), a carrier for the TLR-ligands, were used as a control for pIC+R848 containing treatments.
    pIC+R848
    suggested: None
    Software and Algorithms
    SentencesResources
    Cq value in each well was also automatically calculated by the Maestro 1.1 software based on the threshold baseline.
    Maestro
    suggested: (Maestro, RRID:SCR_016748)
    Data generated by the software were exported to a Microsoft Excel (Microsoft Corporation; Redmond, WA) spreadsheet for data processing.
    Microsoft Excel
    suggested: (Microsoft Excel, RRID:SCR_016137)
    Slides were viewed and digitized pictures were taken by Olympus VS120 scanner and analyzed by a senior Pathologist using Olyvia 3.2 software (Olympus Corporation, Tokyo, Japan).
    Olyvia
    suggested: (OlyVIA , RRID:SCR_016167)
    Results were analyzed with CiraSoft software (Quanterix).
    CiraSoft
    suggested: None
    The reads were quality filtered, trimmed and demultiplexed and aligned to the BioSpyder Human Whole Transcriptome v2.0 annotated reference using Salmon (v1.1.0)(Patro et al., 2017).
    Salmon
    suggested: (Salmon, RRID:SCR_017036)
    Differential expression analysis was conducted using DESeq2 (v1.26.1)(Love et al., 2014) in R (v 3.6.2).
    DESeq2
    suggested: (DESeq, RRID:SCR_000154)
    Principal component analysis (PCA) was performed to determine overall relatedness of the samples under the various experimental conditions using ggplot2 (v 3.3.3) (Wickham, 2016)
    ggplot2
    suggested: (ggplot2, RRID:SCR_014601)
    Pathway analysis was performed using ROntoTools (v 2.14) (Voichita et al., 2019) with a KEGG pathway (Kanehisa, 2019; Kanehisa et al., 2021; Kanehisa and Goto, 2000) considered significantly changed with a false discovery rate perturbation value of > 0.1.
    ROntoTools
    suggested: None
    KEGG
    suggested: (KEGG, RRID:SCR_012773)

    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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).


    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.
    • No funding statement was detected.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.


    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.