Preexisting antibodies targeting SARS-CoV-2 S2 cross-react with commensal gut bacteria and impact COVID-19 vaccine induced immunity
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SciScore for 10.1101/2021.07.13.21260404: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: Experiments using mice and samples of healthy human were approved by the Research Ethics Review Committee of the Shanghai Public Health Clinical Center Affiliated to Fudan University. Sex as a biological variable Mouse vaccination: Peripheral blood samples were collected from female adult mice and pre-existing S2 binding antibodies were measured using the previously described in-house ELISA method. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Detection of SARS-CoV-2 S1 and S2 specific binding antibodies: In-house enzyme-linked immunosorbent assays (ELISA) were developed to … SciScore for 10.1101/2021.07.13.21260404: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IRB: Experiments using mice and samples of healthy human were approved by the Research Ethics Review Committee of the Shanghai Public Health Clinical Center Affiliated to Fudan University. Sex as a biological variable Mouse vaccination: Peripheral blood samples were collected from female adult mice and pre-existing S2 binding antibodies were measured using the previously described in-house ELISA method. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Detection of SARS-CoV-2 S1 and S2 specific binding antibodies: In-house enzyme-linked immunosorbent assays (ELISA) were developed to measure SARS-CoV-2 S1 and S2 specific binding antibodies. SARS-CoV-2suggested: NoneS1suggested: NoneThen, 100μl of a HRP labeled goat anti-mouse IgG antibody (Cat# ab6759, Abcam, UK) or goat anti-mouse IgG antibody (Cat# 115-035-003, Jackson Immuno Research, USA) diluted in 1×PBS containing 5% milk were added to each well and incubated for 1 hour at 37°C. anti-mouse IgGsuggested: (Abcam Cat# ab6759, RRID:AB_955434)Preparation of P144 specific monoclonal antibodies: Monoclonal antibodies against P144 were prepared from one naïve BALB/c mouse and one naïve C57BL/6 mouse respectively using the hybridoma technique. P144suggested: NoneBriefly, the 96-well ELISPOT plates were coated with purified anti-mouse IFN-γ monoclonal antibody overnight at 4°C. anti-mouse IFN-γsuggested: NoneAfter incubation, detecting antibody and Avidin-HRP were added sequentially. Avidin-HRPsuggested: NoneExperimental Models: Cell Lines Sentences Resources Briefly, freshly isolated splenocytes were mixed and fused with SP2/0 cells at a ratio of 1:10. SP2/0suggested: CLS Cat# 400481/p8829_Sp20-Ag14, RRID:CVCL_2199)Next, Vero cells were added into each well (2×104 cells/well) and the plates were incubated at 37°C in a humidified incubator with 5% CO2. Verosuggested: RRID:CVCL_ZW93)Experimental Models: Organisms/Strains Sentences Resources Preparation of P144 specific monoclonal antibodies: Monoclonal antibodies against P144 were prepared from one naïve BALB/c mouse and one naïve C57BL/6 mouse respectively using the hybridoma technique. BALB/csuggested: NoneC57BL/6suggested: NoneSelected clones of hybridoma cells were injected intraperitoneally into BALB/c×ICR hybrid mice. BALB/c×ICRsuggested: RRID:MGI:5652525)Recombinant DNA Sentences Resources The codon optimized spike gene was subcloned into a eukaryotic expression vector (pJW4303, kindly gifted by Dr. Shan Lu’s Laboratory at the University of Massachusetts) (35). pJW4303suggested: NoneSoftware and Algorithms Sentences Resources The data were analyzed using the FlowJo software (BD Biosciences, USA) FlowJosuggested: (FlowJo, RRID:SCR_008520)Construction and preparation of a candidate DNA vaccine encoding SARS-CoV-2 full length S protein: The full-length s gene sequence of the reference SARS-CoV-2 strain was optimized according to the preference of human codon usage and synthesized by GENEWIZ life science company (Suchow, China). GENEWIZsuggested: (GENEWIZ, RRID:SCR_003177)Statistical analysis: All statistical analyses were performed using GraphPad Prism 8 (GraphPad Software, Inc., La Jolla, CA, USA). GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)GraphPadsuggested: (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: Please consider improving the rainbow (“jet”) colormap(s) used on page 41. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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.
Results from scite Reference Check: We found no unreliable references.
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