Human angiotensin-converting enzyme 2 transgenic mice infected with SARS-CoV-2 develop severe and fatal respiratory disease

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1. Version published to 10.1172/jci.insight.142032

   Oct 2, 2020
2. 

   ScreenIT

   Jul 12, 2020

   SciScore for 10.1101/2020.07.09.195230: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Animal experimental protocols were approved by a standing internal 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
   Then the sections were incubated with primary antibodies: rabbit polyclonal anti-SARS-CoV Spike at a dilution of 1:200 (40150-T62-COV2, Sino Biological, Chesterbrook, PA, USA), mouse monoclonal anti-SARS-CoV NP at a dilution of 1:200 (40143-MM05, Sino Biological), mouse monoclonal anti-pan cytokeratin at a dilution of 1:100 (sc-8018, Santa Cruz …

   SciScore for 10.1101/2020.07.09.195230: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Animal experimental protocols were approved by a standing internal 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
   Then the sections were incubated with primary antibodies: rabbit polyclonal anti-SARS-CoV Spike at a dilution of 1:200 (40150-T62-COV2, Sino Biological, Chesterbrook, PA, USA), mouse monoclonal anti-SARS-CoV NP at a dilution of 1:200 (40143-MM05, Sino Biological), mouse monoclonal anti-pan cytokeratin at a dilution of 1:100 (sc-8018, Santa Cruz Biotechnology, Dallas, TX, USA), mouse monoclonal anti-e-cadherin at a dilution of 1:100 (33-4000, Thermo Fisher Scientific, Waltham, MA, USA), rabbit polyclonal anti-myeloperoxidase (MPO) at a dilution of 1:200 (A039829-2, Dako Agilent Pathology Solutions, Carpinteria, CA, USA), rabbit polyclonal anti-CD3 antibody at a dilution of 1:200 (A045229-2, Dako Agilent Pathology Solutions), rat monoclonal anti-CD45 antibody at a dilution of 1:100 (05-1416, Millipore Sigma, Burlington, MA, USA), rabbit polyclonal anti-CD68 at a dilution of 1:200 (ab125212, Abcam, Cambridge, MA, USA), mouse monoclonal anti-NeuN at a dilution of1:200 (MAB377, Millipore Sigma), and/or chicken polyclonal anti-GFAP at a dilution of 1:200 (ab4674, Abcam) for 2 hours at room temperature.	anti-SARS-CoV suggested: (Rockland Cat# 200-401-A51, RRID:AB_828457) anti-SARS-CoV NP suggested: (RayBiotech Cat# MD-05-0425, RRID:AB_951740) anti-pan cytokeratin suggested: (Santa Cruz Biotechnology Cat# sc-8018, RRID:AB_627396) sc-8018 suggested: (Santa Cruz Biotechnology Cat# sc-8018, RRID:AB_627396) anti-e-cadherin suggested: (Thermo Fisher Scientific Cat# 13-1800, RRID:AB_2533004) anti-myeloperoxidase (MPO suggested: (MBL International Cat# JM-3831-100, RRID:AB_1279154) anti-CD3 suggested: (Agilent Cat# A0452, RRID:AB_2335677) anti-CD45 suggested: (Millipore Cat# 05-1416, RRID:AB_10562966) anti-CD68 suggested: (Abcam Cat# ab125212, RRID:AB_10975465) anti-NeuN suggested: (Millipore Cat# MAB377, RRID:AB_2298772) anti-GFAP suggested: (Abcam Cat# ab4674, RRID:AB_304558)
   After rinses with PBT, the sections were incubated with secondary goat anti-rabbit or anti-chicken Alexa Fluor 488 at dilution of 1:500 (Thermo Fisher Scientific) and goat anti-mouse or anti-rat Alexa Fluor 568 at a dilution of 1:500 (Thermo Fisher Scientific) antibodies, for 1 hour at room temperature.	anti-rabbit suggested: None anti-chicken suggested: (Thermo Fisher Scientific Cat# A-11041, RRID:AB_2534098) anti-mouse suggested: None anti-rat suggested: (Thermo Fisher Scientific Cat# A-11077, RRID:AB_2534121)
   Experimental Models: Cell Lines
   Sentences	Resources
   A master challenge stock of virus was propagated by making two passages in Vero76 cells in Modified Eagles Medium with Earle’s Salts (EMEM) (Corning) supplemented with 1% GlutaMAX, 1% NEAA, and 10% heat inactivated fetal bovine serum (FBS) (Gibco).	Vero76 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Mice: C57BL/6J (BL6), Rag2 KO mice and K18-hACE2 mice [B6.Cg-Tg(K18-hACE2)2Prlmn/J] (6-8 weeks old) were purchased from the Jackson Laboratory.	C57BL/6J suggested: None Rag2 KO suggested: RRID:NSRRC_0035) K18-hACE2 suggested: RRID:IMSR_GPT:T037657) B6.Cg-Tg(K18-hACE2)2Prlmn/J suggested: None
   Software and Algorithms
   Sentences	Resources
   Images were captured on a Zeiss LSM 880 confocal system (Zeiss, Oberkochen, Germany) and processed using ImageJ software (National Institutes of Health, Bethesda, MD).	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   Data from each NanoString panel were first processed independently using nSolver (v.4.0) software (NanoString) as follows: following quality control checks on the individual RCC files, raw counts across samples were normalized to the mean counts for spiked synthetic DNA-positive controls present in the hybridization reactions to mitigate platform-associated sources of variation.	nSolver suggested: None
   Candidate reference genes were selected using the nCounter advanced analysis (nCAA) module (v.2.0.115), which implements the geNorm algorithm for downselection(52).	geNorm suggested: (geNORM, RRID:SCR_006763)
   All analyzes were performed using Prism software.	Prism suggested: (PRISM, RRID:SCR_005375)
   Supplemental Materials and Methods:	Methods suggested: None

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   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).

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   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.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 33 and 37. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

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   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.

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3. 

   Version published to 10.1101/2020.07.09.195230v1 on bioRxiv

   Jul 9, 2020