Comprehensive analysis of disease pathology in immunocompetent and immunocompromised hamster models of SARS-CoV-2 infection

  1. Santhamani Ramasamy
  2. Afsal Kolloli
  3. Ranjeet Kumar
  4. Seema Husain
  5. Patricia Soteropoulos
  6. Theresa L. Chang
  7. Selvakumar Subbian

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Abstract

The pathogenesis of SARS-CoV-2 in the context of a specific immunological niche is not fully understood. Here, we used a golden Syrian hamster model to systematically evaluate the kinetics of host response to SARS-CoV-2 infection, following disease pathology, viral loads, antibody responses, and inflammatory cytokine expression in multiple organs. The kinetics of SARS-CoV-2 pathogenesis and genomewide lung transcriptome was also compared between immunocompetent and immunocompromised hamsters. We observed that the body weight loss was proportional to the SARS-CoV-2 infectious dose and lasted for a short time only in immunocompetent hamsters. Body weight loss was more prominent and prolonged in infected immunocompromised hamsters. While the kinetics of viral replication and peak live viral loads were not significantly different at low and high infectious doses (LD and HD), the HD-infected immunocompetent animals developed severe lung disease pathology. The immunocompetent animals cleared the live virus in all tested tissues by 12 days post-infection and generated a robust serum antibody response. In contrast, immunocompromised hamsters mounted an inadequate SARS-CoV-2 neutralizing antibody response, and the virus was detected in the pulmonary and multiple extrapulmonary organs until 16 days post-infection. These hamsters also had prolonged moderate inflammation with severe bronchiolar-alveolar hyperplasia/metaplasia. Consistent with the difference in disease presentation, distinct changes in the expression of inflammation and immune cell response pathways and network genes were seen in the lungs of infected immunocompetent and immunocompromised animals. This study highlights the interplay between the kinetics of viral replication and the dynamics of SARS-CoV-2 pathogenesis at organ-level niches and maps how COVID-19 symptoms vary in different immune contexts. Together, our data suggest that the histopathological manifestations caused by progressive SARS-CoV-2 infection may be a better predictor of COVID-19 severity than individual measures of viral load, antibody response, and cytokine storm at the systemic or local (lungs) levels in the immunocompetent and immunocompromised hosts.

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    SciScore for 10.1101/2022.01.07.475406: (What is this?)

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

    Table 1: Rigor

    EthicsIACUC: All animal procedures were performed in bio-safety level 3 facilities following procedures approved by the Rutgers University Institutional Animal Care and Use Committee, which is consistent with the policies of the American Veterinary Medical Association (AVMA), the Center for Disease Control (CDC) and the United States Department of Agriculture (USDA). 4.4.
    Sex as a biological variableGolden Syrian hamster infection and sample collection: Seventy-five (n=75) male Golden Syrian hamsters (Mesocricetusauratus) between 6-8 weeks old were purchased (Envigo corporation, Denver, PA, USA) and housed at two animals/cage.
    Randomizationnot detected.
    BlindingThe identity of samples was blinded before analysis, and histopathological examination was performed by a veterinary pathologist (S.R) using the EVOS FL Cell imaging system (Thermo Fischer Scientific, USA).
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The sections were blocked using 1% BSA for 1 hr and incubated with rabbit anti-hamster ACE2 antibody (Product no. HPA000288, Millipore Sigma, Burlington, MA, USA) overnight at 4°C.
    anti-hamster ACE2
    suggested: None
    Alexa-488 labeled anti-rabbit secondary antibodies (Cat no. ab150077, Abcam, MA, USA) prepared at 1:2000 dilution in 5% BSA were used to detect ACE2 expression.
    anti-rabbit
    suggested: (Abcam Cat# ab150077, RRID:AB_2630356)
    Experimental Models: Cell Lines
    SentencesResources
    Virus and cell lines: SARS-CoV-2 (strain USA-WA1/2020) infected Vero E6 cell lysate was obtained from BEI Resources (BEI Resources, Manassas, VA, USA).
    Vero E6
    suggested: RRID:CVCL_XD71)
    Software and Algorithms
    SentencesResources
    Histopathology images were organized and labeled using Adobe Photoshop v22.1.1 and Adobe Illustrator v25.1.
    Adobe Photoshop
    suggested: (Adobe Photoshop, RRID:SCR_014199)
    Adobe Illustrator
    suggested: (Adobe Illustrator, RRID:SCR_010279)
    Spleen white pulp cells were quantified manually and using ImageJ (NIH, Bethesda, USA).
    ImageJ
    suggested: (ImageJ, RRID:SCR_003070)
    Images were captured using Axiovert 200M inverted fluorescence microscope (Zeiss, Oberkochen, Germany) using 20X or 63X oil-immersion objective with Prime sCMOS camera (Photometrics, Tucson, AZ) by Metamorph image acquisition software (Molecular Devices, San Jose, CA) ImageJ software was used for analyses.
    Metamorph
    suggested: None
    The SDEG were also analyzed by using Ingenuity Pathway Analysis (IPA) software (Ingenuity® Systems, Inc. Redwood City, CA) to determine the networks and pathways that are affected in SARS-CoV-2 infected and/or Uninfected hamster lungs with or without immunosuppression at 4 dpi and 16dpi, as described previously 61, 62.
    Ingenuity Pathway Analysis
    suggested: (Ingenuity Pathway Analysis, RRID:SCR_008653)
    Statistical analysis: Statistical analysis was performed using GraphPad Prism-8 (GraphPad Software, La Jolla, CA), and the mean ± standard error (SE) values were plotted as graphs.
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    The RNAseq data has been submitted to Gene Expression Omnibus (GEO) of the NCBI (accession number awaited).
    Gene Expression Omnibus
    suggested: (Gene Expression Omnibus (GEO, RRID:SCR_005012)

    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: Please consider improving the rainbow (“jet”) colormap(s) used on page 55. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

    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.

    About SciScore

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  2. Version published to 10.1101/2022.01.07.475406v1 on bioRxiv