HDL-scavenger receptor B type 1 facilitates SARS-CoV-2 entry

  1. Congwen Wei
  2. Luming Wan
  3. Qiulin Yan
  4. Xiaolin Wang
  5. Jun Zhang
  6. Xiaopan Yang
  7. Yanhong Zhang
  8. Chen Fan
  9. Dongyu Li
  10. Yongqiang Deng
  11. Jin Sun
  12. Jing Gong
  13. Xiaoli Yang
  14. Yufei Wang
  15. Xuejun Wang
  16. Jianmin Li
  17. Huan Yang
  18. Huilong Li
  19. Zhe Zhang
  20. Rong Wang
  21. Peng Du
  22. Yulong Zong
  23. Feng Yin
  24. Wanchuan Zhang
  25. Nan Wang
  26. Yumeng Peng
  27. Haotian Lin
  28. Jiangyue Feng
  29. Chengfeng Qin
  30. Wei Chen
  31. Qi Gao
  32. Rui Zhang
  33. Yuan Cao
  34. Hui Zhong

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Abstract

No abstract available

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  1. Version published to 10.1038/s42255-020-00324-0
  2. ScreenIT

    SciScore for 10.1101/2020.08.13.248872: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line AuthenticationContamination: All cell lines were previously tested for mycoplasma contamination and incubated in Dulbecco’s modified Eagle’s medium at 37 °C under a humidified atmosphere with 5% CO2.

    Table 2: Resources

    Antibodies
    SentencesResources
    Reagents and antibodies: His-tagged SARS-2-S (S1+S2 ECD; YP_009724390.1;Val16-Pro1213; 40589-V08B1), SARS-2-S1 (YP_009724390.1;Val16-Arg685; 40591-V08H), and SARS-2-S2 (ECD; YP_009724390.1; Ser686-Pro1213; 40591-V08B) proteins were from Sino Biological.
    S1+S2
    suggested: None
    ECD
    suggested: None
    YP_009724390.1;Val16-Pro1213
    suggested: None
    40589-V08B1
    suggested: None
    SARS-2-S1
    suggested: None
    YP_009724390.1;Val16-Arg685
    suggested: None
    40591-V08H
    suggested: None
    SARS-2-S2
    suggested: None
    An antibody against ACE2 for FACS (21115-1-AP; 1:100 dilution) was from Proteintech.
    21115-1-AP
    suggested: (Proteintech Cat# 21115-1-AP, RRID:AB_10732845)
    A PE anti-His antibody (362603, clone J095G46; 1;100 dilution) and an antibody against SR-B1 for FACS (363208, clone m1B9; dilution 1:100) were from Biolegend.
    anti-His
    suggested: (BioLegend Cat# 362603, RRID:AB_2563634)
    SR-B1 for FACS ( 363208
    suggested: None
    The SARS-CoV-2 S1 antibody (40150-R007) for confocal microscopy was from Sino Biological.
    S1
    suggested: None
    Then, anti-rabbit or anti-mouse HRP-conjugated antibodies were applied after 3 washes and the antigen-antibody complexes were visualized using chemilumines cence.
    anti-rabbit
    suggested: None
    anti-mouse HRP-conjugated
    suggested: None
    Flow cytometry for surface SR-B1 or ACE2 expression analysis: Cells were washed with PBS 2 times and incubated with APC-conjugated (APC anti-SR-B1) antibody (1:100) or anti-ACE2 antibody (1:100) followed by a secondary incubation with donkey-anti-rabbit antibody (1:200) conjugated to Alexa Flour 488.
    antibody
    suggested: None
    After incubating at RT for 30 min, the cells were washed two times with PBS containing 2% FBS and then incubated for 30 min at RT with a PE-conjugated antibody (PE anti-His tag).
    anti-His tag) .
    suggested: None
    After blocking in 1% normal goat serum, the sections were incubated with ACE2 or SR-B1 monoclonal antibodies at 4□°C overnight in a humidified chamber, which was followed by an incubation with HRP-labeled goat anti-mouse IgG secondary antibody (Beijing ZSGB Biotechnology, ZDR-5307).
    ACE2
    suggested: None
    SR-B1
    suggested: None
    anti-mouse IgG
    suggested: None
    Subsequently, the sections were incubated with a goat anti-rabbit IgG secondary antibody (HRP) (Beijing ZSGB Biotechnology, PV9001) for 60 min and then visualized with 3,30-diaminobenzidine tetrahydrochloride (DAB).
    anti-rabbit IgG
    suggested: (ZSGB-Bio Cat# PV-9001, RRID:AB_2868452)
    Experimental Models: Cell Lines
    SentencesResources
    , Vero E6 (CRL-1568), MDCK (CCL-34) and Hepa 1-6 (CRL-1830
    Vero E6
    suggested: None
    MDCK
    suggested: None
    Pseudovirus production: All pseudoparticles were generated in 293T cells transfected with the HIV backbone vector pNL4-3.
    293T
    suggested: None
    For chemicals and antibodies, Vero E6 or Huh-7 cells (1.6 ×105 cells/mL) were cultured in a 96-well plate at 37 °C overnight.
    Huh-7
    suggested: None
    For SR-B1 expression, Vero cells (1.6 ×105 cells/mL) were transfected with a plasmid encoding SR-B1, while Huh-7 cells (1.6 ×105 cells/mL) were transfected with siRNA oligos for SR-B1.
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    Data were fitted to saturation binding equations using GraphPad Prism 6.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Acoustic Focusing cytometer using Flowjo V10.0.7.
    Flowjo
    suggested: (FlowJo, RRID:SCR_008520)
    Statistical methods: In the present study, GraphPad Prism 8.0 was used for statistical calculations and data plotting.
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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.

    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.

  3. Version published to 10.1101/2020.08.13.248872v1 on bioRxiv