The pathogenicity of SARS-CoV-2 in hACE2 transgenic mice

  1. Linlin Bao
  2. Wei Deng
  3. Baoying Huang
  4. Hong Gao
  5. Jiangning Liu
  6. Lili Ren
  7. Qiang Wei
  8. Pin Yu
  9. Yanfeng Xu
  10. Feifei Qi
  11. Yajin Qu
  12. Fengdi Li
  13. Qi Lv
  14. Wenling Wang
  15. Jing Xue
  16. Shuran Gong
  17. Mingya Liu
  18. Guanpeng Wang
  19. Shunyi Wang
  20. Zhiqi Song
  21. Linna Zhao
  22. Peipei Liu
  23. Li Zhao
  24. Fei Ye
  25. Huijuan Wang
  26. Weimin Zhou
  27. Na Zhu
  28. Wei Zhen
  29. Haisheng Yu
  30. Xiaojuan Zhang
  31. Li Guo
  32. Lan Chen
  33. Conghui Wang
  34. Ying Wang
  35. Xinming Wang
  36. Yan Xiao
  37. Qiangming Sun
  38. Hongqi Liu
  39. Fanli Zhu
  40. Chunxia Ma
  41. Lingmei Yan
  42. Mengli Yang
  43. Jun Han
  44. Wenbo Xu
  45. Wenjie Tan
  46. Xiaozhong Peng
  47. Qi Jin
  48. Guizhen Wu
  49. Chuan Qin

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Abstract

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  1. ScreenIT

    SciScore for 10.1101/2020.02.07.939389: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementAll procedures in this study involving animals were reviewed and approved by the Institutional Animal Care and Use Committee of the Institute of Laboratory Animal Science , Peking Union Medical College ( BLL20001)Randomizationnot detected.Blindingnot detected.Power Analysisnot detected.Sex as a biological variableSpecific pathogen-free , 6-11-month-old , male and female WT mice ( WT-HB-01 , n=15 ) and hACE2 mice ( ACE2-HB-01 , n=19 ) were inoculated intranasally with SARSCoV-2 stock virus ( HB-01 ) at a dosage of 105 TCID50/50 Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Moreover , specific IgG antibodies against S protein of SARS-CoV-2 were positively measured in the sera of ACE2-HB-01 mice at 21 dpi ( Figure 1e) .
    specific IgG
    suggested: None
    Alternatively , the sections were stained with MAC2 antibody ( Cedarlane Laboratories , CL8942AP)
    MAC2
    suggested: (CEDARLANE Cat# CL8942AP, AB_10060357)
    , CD3 antibody ( Dako , A0452 ) or CD19 antibody ( Cell Signaling Technology , 3574 ) at 4 ℃ overnight .
    CD3
    suggested: (Agilent Cat# A0452, AB_2335677)
          <div style="margin-bottom:8px">
        <div><b>CD19</b></div>
        <div>suggested: (Cell Signaling Technology Cat# 3574, <a href="https://scicrunch.org/resources/Any/search?q=AB_2275523">AB_2275523</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Subsequently , the sections were goat anti-rat IgG secondary antibody ( HRP ) ( Beijing ZSGB Biotechnology , PV9004)</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-rat IgG</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>PV9004</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">goat anti-rabbit IgG secondary antibody ( HRP ) ( Beijing ZSGB Biotechnology , PV9001 ) for 60 min , and visualized by incubation with 3,30-diaminobenzidine tetrahydrochloride ( DAB) .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-rabbit IgG</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Anti-S protein antibody ( mouse monoclonal 7D2 , laboratory preparation , 1:200 ) and anti-hACE2 antibody ( rabbit polyclonal , ab15348 , Abcam1:200 ) hACE2 was used as the positive control .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Anti-S protein antibody</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-hACE2</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">For the viral antigen , the sections from ACE2Mock mice incubated with anti-S protein antibody were used as the negative control .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-S protein</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Co-localization of SARS-CoV-2 S protein and hACE2 receptor in hACE2 mouse lungs, the sections were incubated with anti-SARS-CoV-2 S protein antibody, anti-human ACE2 antibody, and DAPI.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>anti-SARS-CoV-2 S protein</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-human ACE2</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Identification of 7D2 antibody against SARS-CoV-2 S1 protein.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>SARS-CoV-2 S1 protein.</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The plate coated by 0.2 ug SARS-CoV-2 S1 protein was incubated with 7D2 antibody as primary antibody (1:200) and detected using HRP-conjugated goat antimouse secondary antibody.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>antimouse</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Experimental Models: Cell Lines</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Seed SARS-CoV-2 stocks and virus isolation studies were performed in Vero cells , which are maintained in Dulbecco 's modified Eagle 's medium ( DMEM , Invitrogen , Carlsbad , USA ) supplemented with 10 % fetal bovine serum ( FBS) , 100 IU/ml penicillin , and 100 µg/ml streptomycin , and incubated at 37°C , 5 % CO2 .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Vero</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Transmission Electron Microscopy Supernatant from Vero E6 cell cultures that showed cytopathic effects was collected , inactivated with 2 % paraformaldehyde for at least 2 hours , and ultracentrifuged to sediment virus particles .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Vero E6</b></div>
        <div>suggested: <a href="https://scicrunch.org/resources/Any/search?q=CVCL_XD71">CVCL_XD71</a></div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Software and Algorithms</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The speed of geographical spread of COVID-19 caused by SARS-CoV-2 has been declared as public health emergency of international concern ( PHEIC) , with cases reported on multiple continents only weeks after the disease was first reported6 .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>SARS-CoV-2</b></div>
        <div>suggested: (Sino Biological Cat# 40143-R019, <a href="https://scicrunch.org/resources/Any/search?q=AB_2827973">AB_2827973</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Statistical analysis All data were analyzed with GraphPad Prism 8.0 software.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>GraphPad Prism</b></div>
        <div>suggested: (GraphPad Prism, <a href="https://scicrunch.org/resources/Any/search?q=SCR_002798">SCR_002798</a>)</div>
      </div>
    </td></tr></table>

    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 is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.

  2. Version published to 10.1038/s41586-020-2312-y
  3. Version published to 10.1101/2020.02.07.939389v3 on bioRxiv
  4. Version published to 10.1101/2020.02.07.939389v2 on bioRxiv
  5. Version published to 10.1101/2020.02.07.939389v1 on bioRxiv