Dynamics of humoral and cellular immune responses after homologous and heterologous SARS-CoV-2 vaccination with ChAdOx1 nCoV-19 and BNT162b2

Article activity feed

1. Version published to 10.1016/j.ebiom.2022.104294

   Nov 1, 2022
2. 

   ScreenIT

   Mar 29, 2022

   SciScore for 10.1101/2022.03.23.22272771: (What is this?)

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

   Table 1: Rigor

   Ethics	IRB: Ethics approval was granted by the local ethics committees in Erlangen (Az. 340_21B) and Munich (Az. 26/21 and Az.
   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
   Study design and participants: The study is a follow-up analysis of 473 homologously or heterologously vaccinated participants that were previously only assessed for the production of antibodies using sVNT5.	sVNT5 suggested: None
   Antibody avidity: Binding strength of the SARS-Cov-2 IgG antibodies was determined by adaptation of the commercial IgG agile SARS-CoV-2 ELISA …

   SciScore for 10.1101/2022.03.23.22272771: (What is this?)

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

   Table 1: Rigor

   Ethics	IRB: Ethics approval was granted by the local ethics committees in Erlangen (Az. 340_21B) and Munich (Az. 26/21 and Az.
   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
   Study design and participants: The study is a follow-up analysis of 473 homologously or heterologously vaccinated participants that were previously only assessed for the production of antibodies using sVNT5.	sVNT5 suggested: None
   Antibody avidity: Binding strength of the SARS-Cov-2 IgG antibodies was determined by adaptation of the commercial IgG agile SARS-CoV-2 ELISA (Virion/Serion, Germany) using ammonium thiocyanate (NH4SCN) (Roth, Germany) as previously described16,54,55.	SARS-Cov-2 IgG suggested: None
   Serum samples were measured using the IgG agile SARS-CoV-2 ELISA and diluted to 100 U/mL according to the standard curve provided by the manufacturer to exclude an influence of variable antibody concentrations.	SARS-CoV-2 suggested: None ELISA suggested: None
   As a primary antibody, the SinoBiological anti-SARS-CoV-2-N T62 antibody (40143-T62) was used.	anti-SARS-CoV-2-N T62 suggested: None
   Goat anti-rabbit IgG2a-HRP antibody (EMD Millipore / order number 12-348) with 1% FCS-PBS was diluted to 1:4000 ratio.	Goat anti-rabbit IgG2a-HRP antibody suggested: None anti-rabbit IgG2a-HRP suggested: None
   FACS-based analysis of anti-S binding antibodies: A modified version of our previously published serological assay was used, in which HEK 293T cells either stably expressing the spike protein from the original Wuhan strain or transiently expressing the spike protein of B.1.167.2 or B1.1.529, respectively, were used as target cells56.	anti-S suggested: None
   To quantify antigen-specific antibodies, 5×105 HEK 293T cells were incubated with serum samples diluted in 100 µl FACS-PBS (PBS with 0.5% BSA and 1 mM sodium azide) for 20 minutes at 4°C to bind to spike protein on the surface.	antigen-specific suggested: (Miltenyi Biotec Cat# 130-092-105, RRID:AB_615062)
   After washing with 200 µl buffer, bound S-specific antibodies were detected with anti-human IgG-AF647 (4°C, 30 min incubation; clone HP6017, Biolegend, Cat #409320).	anti-human IgG-AF647 suggested: (SouthernBiotech Cat# 2040-31, RRID:AB_2795651)
   The median fluorescence intensity (MFI) correlates with the level of bound antibodies56.	antibodies56 suggested: None
   ELISPOT plates (Millipore) were coated with anti-human IFN-γ monoclonal antibody (clone 1-DIK, Mabtech) at 0.5 µg/well overnight at 4°C.	anti-human IFN-γ suggested: None
   After the stimulation period, the plates were washed and 80 µL of anti-human IFN-γ (FITC)/anti-human IL-2 (Hapten2) detection antibody solution was added for 2h at room temperature.	Hapten2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   After one hour of preincubation, the inoculum was transferred to the pre-seeded VeroE6 cells for another one-hour incubation at 37°C before the inoculum was replaced by supplemented DMEM.	VeroE6 suggested: None
   FACS-based analysis of anti-S binding antibodies: A modified version of our previously published serological assay was used, in which HEK 293T cells either stably expressing the spike protein from the original Wuhan strain or transiently expressing the spike protein of B.1.167.2 or B1.1.529, respectively, were used as target cells56.	HEK 293T suggested: None
   For the assessment of pseudotype neutralization, HEK293T-ACE2 cells were seeded at 2×104 cells/well in a 96well flat bottom plate.	HEK293T-ACE2 suggested: None
   Recombinant DNA
   Sentences	Resources
   To produce pseudotyped reporter particles, HEK293T cells were transfected with the SIV-based self-inactivating vector encoding luciferase (pGAE-LucW), the SIV-based packaging plasmid (pAdSIV3), and the respective spike variant-encoding plasmid as described previously58.	pGAE-LucW suggested: None
   Software and Algorithms
   Sentences	Resources
   The inhibition curve of each sample was analyzed by statistical analysis software Graph Pad Prism (GraphPad Software, USA), and 50% inhibitory concentration (IC50) was determined using non-linear regression.	Graph Pad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   After further washing, samples were measured on an AttuneNxt (ThermoFisher) and analyzed using FlowJo software (Tree Star Inc.)	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   The reciprocal serum ID50 was determined with Prism GraphPad 9 (San Diego, California, USA) by application of the Sigmoidal 4PL function.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   Isolation and cultivation of peripheral blood mononuclear cell (PBMC): PBMCs were isolated from citrate peripheral blood of vaccinated individuals by density gradient centrifugation using Biocoll® separating solution, density 1.077 g/ml (Bio&Sell) and frozen in heat-inactivated FCS + 10% DMSO (Sigma-Aldrich) for liquid nitrogen storage.	Biocoll® suggested: None
   Statistical analysis and graphical presentation: Statistics as well as figures were created with PRISM GraphPad 9.3.1.	PRISM GraphPad 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.

   ---

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

   ---

   Read the original source
3. 

   Version published to 10.1101/2022.03.23.22272771v1 on medRxiv

   Mar 27, 2022