The N501Y mutation in SARS-CoV-2 spike leads to morbidity in obese and aged mice and is neutralized by convalescent and post-vaccination human sera

Article activity feed

1. 

   ScreenIT

   Jan 21, 2021

   SciScore for 10.1101/2021.01.19.21249592: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Mouse models for comorbidities: All mice strains were obtained from Charles River Laboratories, MA and were housed in a pathogen-free facility at Icahn School of Medicine at Mount Sinai, with food and water ad libitum, adhering to the guidelines from Institutional Animal Care and Use Committee (IACUC).
   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
   Horse radish peroxidase (HRP)-conjugated anti-mouse IgG antibody was purchased from Abcam (ab6823).	anti-mouse IgG antibody suggested: None
   Anti-mouse SARS-CoV-2 …

   SciScore for 10.1101/2021.01.19.21249592: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Mouse models for comorbidities: All mice strains were obtained from Charles River Laboratories, MA and were housed in a pathogen-free facility at Icahn School of Medicine at Mount Sinai, with food and water ad libitum, adhering to the guidelines from Institutional Animal Care and Use Committee (IACUC).
   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
   Horse radish peroxidase (HRP)-conjugated anti-mouse IgG antibody was purchased from Abcam (ab6823).	anti-mouse IgG antibody suggested: None
   Anti-mouse SARS-CoV-2 NP (NP1C7C7) and anti-mouse SARS-CoV-2 spike (2BCE5) antibodies were obtained from Center for Therapeutic Antibody Development at Icahn School of Medicine at Mount Sinai, New York.	Anti-mouse SARS-CoV-2 NP suggested: None anti-mouse SARS-CoV-2 spike (2BCE5) suggested: None
   Antibodies used in Western blot were anti-mouse-ACE-2 (R&D, MAB3437), anti-beta-tubulin (Sigma Aldrich, T8328) and anti-mouse (HRP-conjugated, KwikQuant).	anti-mouse-ACE-2 suggested: None anti-beta-tubulin suggested: (Sigma-Aldrich Cat# T8328, RRID:AB_1844090) anti-mouse suggested: None
   The plaques were immune-stained with anti-mouse SARS-CoV-2-NP and anti-mouse SARS-CoV-2-spike antibodies for 1 hour at RT and consequently with HRP-conjugated anti-mouse secondary IgG antibody for 1 hour at room temperature (RT).	anti-mouse SARS-CoV-2-NP suggested: None anti-mouse SARS-CoV-2-spike suggested: None anti-mouse secondary IgG suggested: None
   Anti-Tubulin and anti-mACE-2 (R&D Systems, Cat# MAB3437) primary antibodies were used at dilution of 1:1000 while secondary HRP-conjugated antibodies were used at dilutions of 1:10000 in 3% BSA-containing TBST. 50% tissue culture infective dose (TCID50) calculation and in vitro micro-neutralization assay: To estimate the neutralizing efficiency of sera from vaccinated or SARS-CoV-2-infected mice or humans, in vitro microneutralization assays were performed similarly to what is described previously (13).	Anti-Tubulin suggested: (LSBio (LifeSpan Cat# LS-C48646-100, RRID:AB_1242672) anti-mACE-2 suggested: None TCID50 suggested: None
   After blocking, the cells were incubated with anti-SARS-CoV-2 NP and anti-spike monoclonal antibodies, mixed in 1:1 ratio, for 1.5 hours at room temperature.	anti-SARS-CoV-2 NP suggested: None anti-spike suggested: None
   The cells were washed in 1xPBS and incubated with 1:5000 diluted HRP-conjugated anti-mouse IgG secondary antibody for 1 hour at RT followed by a brief PBS wash.	anti-mouse IgG suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   A variant of virus (termed MA-SARS-CoV-2) was obtained after series of passaging in different backgrounds of laboratory mice as well as mACE-2 expressing VeroE6 cells.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   For TCID50 calculation, the virus stock was serially diluted 10-fold starting with 1:10 dilution and incubated on Vero-E6 cells for 48 hours followed by fixation in 4% Formaldehyde.	Vero-E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   The mouse adaptation of the SARS-CoV-2 variant was studied in C57Bl6, BALB/c and 129S1/SVMJ (termed 129 for simplicity in the text and figures) mice models.	C57Bl6 suggested: None BALB/c suggested: None

   ---

   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.

   ---

   Read the original source
2. 

   Version published to 10.1101/2021.01.19.21249592v1 on medRxiv

   Jan 20, 2021