No evidence for basigin/CD147 as a direct SARS-CoV-2 spike binding receptor
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Abstract
The spike protein of SARS-CoV-2 is known to enable viral invasion into human cells through direct binding to host receptors including ACE2. An alternate entry receptor for the virus was recently proposed to be basigin/CD147. These early studies have already prompted a clinical trial and multiple published hypotheses speculating on the role of this host receptor in viral infection and pathogenesis. Here, we report that we are unable to find evidence supporting the role of basigin as a putative spike binding receptor. Recombinant forms of the SARS-CoV-2 spike do not interact with basigin expressed on the surface of human cells, and by using specialized assays tailored to detect receptor interactions as weak or weaker than the proposed basigin-spike binding, we report no evidence for a direct interaction between the viral spike protein to either of the two common isoforms of basigin. Finally, removing basigin from the surface of human lung epithelial cells by CRISPR/Cas9 results in no change in their susceptibility to SARS-CoV-2 infection. Given the pressing need for clarity on which viral targets may lead to promising therapeutics, we present these findings to allow more informed decisions about the translational relevance of this putative mechanism in the race to understand and treat COVID-19.
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SciScore for 10.1101/2020.07.25.221036: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibody staining was performed with a similar procedure, except during the first 30 minutes on ice cells were incubated with 30 μg/mL of monoclonal Ab-1 anti-BSG antibody12, then resuspended in 1:500 anti-human IgG antibody conjugated to Cy3 (Sigma anti-BSGsuggested: Noneanti-human IgGsuggested: NoneAnti-human basigin monoclonal antibodies were added at the following concentrations: 1.7 μg/mL Ab-1 (Zenonos et al., formerly known as ch6D9)12, 2.2 μg/mL … SciScore for 10.1101/2020.07.25.221036: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Antibody staining was performed with a similar procedure, except during the first 30 minutes on ice cells were incubated with 30 μg/mL of monoclonal Ab-1 anti-BSG antibody12, then resuspended in 1:500 anti-human IgG antibody conjugated to Cy3 (Sigma anti-BSGsuggested: Noneanti-human IgGsuggested: NoneAnti-human basigin monoclonal antibodies were added at the following concentrations: 1.7 μg/mL Ab-1 (Zenonos et al., formerly known as ch6D9)12, 2.2 μg/mL MEM-M6/1 (Abcam #ab666), and 1.3 μg/mL MEM-M6/6 (Abcam #ab119114)25. Anti-human basiginsuggested: NoneBoth secondary antibodies were conjugated to alkaline phosphatase. alkaline phosphatase.suggested: NoneA control mouse anti-rat Cd4 domain 3+4 monoclonal antibody (OX68) against the tags of our recombinant proteins was used at a 1.6 μg/mL concentration. anti-rat Cd4 domainsuggested: NoneAfter 1 hour of incubation with the primary antibody and three HBS-T washes, secondary antibody was added as 1:7000 donkey anti-human IgG (Abcam #ab102407) for ch6D9 and for all other antibodies as 1:3500 goat anti-mouse IgG (Sigma #A9316). anti-mouse IgGsuggested: (Sigma-Aldrich Cat# A9316, RRID:AB_258446)Experimental Models: Cell Lines Sentences Resources Recombinant protein expression and purification: Human embryonic kidney (HEK)-293E cells were transiently transfected with polyethylenimine as previously described46,47. HEK)-293Esuggested: NoneSoftware and Algorithms Sentences Resources Data processing: For flow cytometry experiments, measurement events for analysis were gated on live singlet cells (based on DAPI and forward and side scatter profiles) using FlowJo version 9. FlowJosuggested: (FlowJo, RRID:SCR_008520)Cytometry data was visualized using the CytoML package50 in R version 3.6.1. CytoMLsuggested: NoneResults 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.
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