Dynamics of humoral and T-cell immunity after three BNT162b2 vaccinations in adults older than 80 years

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1. Version published to 10.1016/s1473-3099(22)00219-5

   May 1, 2022
2. 

   ScreenIT

   Feb 17, 2022

   SciScore for 10.1101/2022.02.10.22270733: (What is this?)

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

   Table 1: Rigor

   Ethics	Consent: All donor provided informed consent to participate in the study. IRB: The study of patients with COVID-19 and vaccinations against COVID-19 was approved by the ethics committee of the medical faculty of the Philipps-University Marburg (study number 40/21-12032021).
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After discarding the supernatants, pellets were resuspended in 50 µL Brilliant Violet Staining Buffer (Biolegend) supplemented with antibodies including anti-CD4-PECy7 (1:400 dilution) for the detection of CD4 T cells, …

   SciScore for 10.1101/2022.02.10.22270733: (What is this?)

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

   Table 1: Rigor

   Ethics	Consent: All donor provided informed consent to participate in the study. IRB: The study of patients with COVID-19 and vaccinations against COVID-19 was approved by the ethics committee of the medical faculty of the Philipps-University Marburg (study number 40/21-12032021).
   Sex as a biological variable	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After discarding the supernatants, pellets were resuspended in 50 µL Brilliant Violet Staining Buffer (Biolegend) supplemented with antibodies including anti-CD4-PECy7 (1:400 dilution) for the detection of CD4 T cells, anti-CD8-BV570 (1:100 dilution), and anti-CD19/123/33-BV510 (all from Biolegend at 1:50,1:50, 1:400 dilution, respectively) for the exclusion of CD8 T cells, monocytes and other myeloid cells, B cells, basophils, and plasmacytoid dendritic cells, and incubated 30 minutes in the dark, at room temperature.	anti-CD4-PECy7 suggested: None anti-CD8-BV570 suggested: None anti-CD19/123/33-BV510 suggested: None
   Cell pellets were then washed twice in 200 µL permeabilization buffer (diluted using Millipore water from 10 x concentrated stock buffer, ThermoFisher, Waltham, MA), and finally resuspended in 50 µL permeabilization buffer supplemented with antibodies targeting CD40L (1:100 dilution conjugated to BV421, Biolegend), and intracellular molecules including anti-IFNγ-R718 (1:100 dilution BD Biosciences, San Jose, CA), and incubated for 30 minutes at room temperature in the dark.	CD40L suggested: (BioLegend Cat# 310823, RRID:AB_10933251) BV421 suggested: (BioLegend Cat# 658709, RRID:AB_2563283) anti-IFNγ-R718 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Following incubation at 37 °C for 1 h, approximately 20,000 Vero C1008 cells (ATCC, Cat. no. CRL-1586, RRID: CVCL_0574) were added.	Vero C1008 detected: (IZSLER Cat# BS CL 87, RRID:CVCL_0574)
   Software and Algorithms
   Sentences	Resources
   FlowJo version 10 (BD, Ashland, OR) and OMIQ.ai (Santa Clara, CA) were used for analyzing flow cytometry data.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Quantification of SARS-CoV-2-specific antibodies: Serum IgG antibodies against the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein were quantified using the automated SARS-CoV-2-IgG-II-Quant-Assay on the Abbott Alinity i analyzer (Abbott, Wiesbaden, Germany), following the manufacturer’s protocol.	Abbott suggested: (Abbott, RRID:SCR_010477)
   Statistical analysis details: Prism version 9 (GraphPad software, San Diego, CA) was used to display data, and perform descriptive statistics and significance testing.	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)

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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: We did not find any issues relating to colormaps.

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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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   Results from scite Reference Check: We found no unreliable references.

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3. 

   Version published to 10.1101/2022.02.10.22270733v1 on medRxiv

   Feb 15, 2022