An immunoPET probe to SARS-CoV-2 reveals early infection of the male genital tract in rhesus macaques

  1. Patrick J. Madden
  2. Yanique Thomas
  3. Robert V. Blair
  4. Sadia Samer
  5. Mark Doyle
  6. Cecily C. Midkiff
  7. Lara A. Doyle-Meyers
  8. Mark E. Becker
  9. Muhammad S. Arif
  10. Michael D. McRaven
  11. Lacy M. Simons
  12. Ann M. Carias
  13. Elena Martinelli
  14. Ramon Lorenzo-Redondo
  15. Judd F Hultquist
  16. Francois J. Villinger
  17. Ronald S. Veazey
  18. Thomas J. Hope

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Abstract

The systemic nature of SARS-CoV-2 infection is highly recognized, but poorly characterized. A non-invasive and unbiased method is needed to clarify whole body spatiotemporal dynamics of SARS-CoV-2 infection after transmission. We recently developed a probe based on the anti-SARS-CoV-2 spike antibody CR3022 to study SARS-CoV-2 pathogenesis in vivo . Herein, we describe its use in immunoPET to investigate SARS-CoV-2 infection of three rhesus macaques. Using PET/CT imaging of macaques at different times post-SARS-CoV-2 inoculation, we track the 64 Cu-labelled CR3022-F(ab’)2 probe targeting the spike protein of SARS-CoV-2 to study the dynamics of infection within the respiratory tract and uncover novel sites of infection. Using this method, we uncovered differences in lung pathology between infection with the WA1 isolate and the delta variant, which were readily corroborated through computed tomography scans. The 64 Cu-CR3022-probe also demonstrated dynamic changes occurring between 1- and 2-weeks post-infection. Remarkably, a robust signal was seen in the male genital tract (MGT) of all three animals studied. Infection of the MGT was validated by immunofluorescence imaging of infected cells in the testicular and penile tissue and severe pathology was observed in the testes of one animal at 2-weeks post-infection. The results presented here underscore the utility of using immunoPET to study the dynamics of SARS-CoV-2 infection to understand its pathogenicity and discover new anatomical sites of viral replication. We provide direct evidence for SARS-CoV-2 infection of the MGT in rhesus macaques revealing the possible pathologic outcomes of viral replication at these sites.

Graphic Abstract

PET/CT detected SARS-CoV-2 infection of 4 different tissues in the male genital tract illuminates the cause of COVID-19 clinical sequalae of male sexual health and fertility Figure 1.

Diagram shows schematic illustration of the male genital tract of the rhesus macaque. Virus icon shows sites of SARS-CoV-2 PET signal. Text highlighting the clinical sequalae associated with each sight of infection is shown in text adjacent to each infection site.

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  1. Version published to 10.1101/2022.02.25.481974v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2022.02.25.481974: (What is this?)

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

    Table 1: Rigor

    EthicsIACUC: All procedures were reviewed and approved by the Tulane University Institutional Animal Use Committee under protocol number P0452.
    Sex as a biological variableThree male rhesus macaques (Macaca mulatta) were used in this study.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Lung sections were blocked with 10% normal goat serum (NGS) for 40 minutes, followed by a 60-minute incubation with the anti-SARS primary antibody diluted in NGS.
    anti-SARS
    suggested: None
    Sequential staining of FFPE testes, for CD206 and Caspase 3, was done as described above with 1% normal donkey serum (NDS) being used in place of NGS for blocking and antibody dilutions.
    CD206
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Virus stock was prepared in Vero E6 cells and sequence confirmed by deep sequencing.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Delta variant stock was expanded using Calu-3 cells and prepared as above using Vero E6 cells.
    Calu-3
    suggested: BCRJ Cat# 0264, RRID:CVCL_0609)
    Software and Algorithms
    SentencesResources
    Cu64 chloride was obtained from Washington University, St. Louis MO and shipped overnight to TNPRC.
    TNPRC
    suggested: None
    Both PCA and agglomerative hierarchical clustering were performed using FactoMineR package and factoextra was used for visualization of the clustering results.
    FactoMineR
    suggested: (FactoMineR, RRID:SCR_014602)

    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.

    About SciScore

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  3. Version published to 10.1101/2022.02.25.481974v1 on bioRxiv