Generation of potent cellular and humoral immunity against SARS-CoV-2 antigens via conjugation to a polymeric glyco-adjuvant
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SciScore for 10.1101/2021.05.20.445060: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Samples were excluded from analysis only when an animal developed a health problem for a nontreatment-related reason, according to the animal care guidelines.
Field Sample Permit: Animals: All studies with animals were carried out in accordance with procedures approved by the Institutional Animal Care and Use Committee at the University of Chicago (protocol # 72551) and housed in a specific pathogen-free environment at the University of Chicago.Sex as a biological variable C57Bl/6 female mice aged 8, 21, 47 or greater than 64 weeks were obtained from The Jackson Laboratory Randomization Animals were randomly assigned to a treatment group, and analyses were performed in a blinded fashion. Bl… SciScore for 10.1101/2021.05.20.445060: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: Samples were excluded from analysis only when an animal developed a health problem for a nontreatment-related reason, according to the animal care guidelines.
Field Sample Permit: Animals: All studies with animals were carried out in accordance with procedures approved by the Institutional Animal Care and Use Committee at the University of Chicago (protocol # 72551) and housed in a specific pathogen-free environment at the University of Chicago.Sex as a biological variable C57Bl/6 female mice aged 8, 21, 47 or greater than 64 weeks were obtained from The Jackson Laboratory Randomization Animals were randomly assigned to a treatment group, and analyses were performed in a blinded fashion. Blinding Animals were randomly assigned to a treatment group, and analyses were performed in a blinded fashion. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In the study, the humoral response in mice vaccinated with Spike-p(Man-TLR7), Spike-p(Man-TLR7)+alum, or RBD-p(Man-TLR7) was characterized by evaluating the antibody titers (IgG and IgA) via ELISA, as well as through a viral peptide array and virus neutralization assay. IgAsuggested: NoneAfter 6 washes with PBS-T, wells were incubated for 1 hour at room temperature with horseradish peroxide (HRP)-conjugated antibody against human IgG (Jackson ImmunoResearch). human IgGsuggested: NoneAnti-RBD and anti-Spike antibody analysis: Blood was collected from vaccinated mice weekly or every two weeks into EDTA-K2-coated tubes (Milian). Anti-RBDsuggested: NoneAfter 6 washes with PBS-T, wells were incubated for 1 hour at room temperature with horseradish peroxide (HRP)-conjugated antibody against mouse IgG, IgG1, IgG2b, IgG2c, IgG3, or IgA (Southern Biotech). mouse IgGsuggested: NoneIgG1suggested: NoneIgG2bsuggested: NoneIgG2csuggested: NoneIgG3suggested: NoneIgA ( Southern Biotech)suggested: NoneAfter 6 washes with PBS-T, bound anti-RBD or anti-Spike antibodies were incubated with tetramethylbenzidine substrate for 18 min. anti-Spikesuggested: NoneExperimental Models: Cell Lines Sentences Resources These mixtures were then applied to Vero-E6 cells and maintained until > 90% cell death occurred in the “no serum” control condition (about 4-5 days). Vero-E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources C57Bl/6 female mice aged 8, 21, 47 or greater than 64 weeks were obtained from The Jackson Laboratory C57Bl/6suggested: NoneSoftware and Algorithms Sentences Resources The band density of the reduced samples and RBD or Spike standard curve was then analyzed using ImageJ and the RBD or Spike concentration of the samples was calculated using the standard curve generated. ImageJsuggested: (ImageJ, RRID:SCR_003070)For AUC analysis, the fold over the median background absorbance was calculated for each sample, and GraphPad Prism (version 8) was then used to calculate the AUC of the log-transformed plot. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Spots were analyzed using Spotfinder software (version v3.2.1). Spotfindersuggested: (Spotfinder, RRID:SCR_000085)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: We detected the following sentences addressing limitations in the study:Despite the advantages of the flexible antigen-adjuvant conjugation chemistry, this is also the source of the primary limitation of the p(Man-TLR7) vaccine platform. Although the conjugation of antigen to the polymer is via a self-immolative linker, we observed that this may lead to reduced activity of the antigen, in terms of recognition of its target receptor (Fig 1C and Fig. S3C). This suggests that the smaller the antigen, the fewer the number of potential epitopes, and the higher the chances that conjugation to p(Man-TLR7) may lead to steric blockade of the receptor-binding site. This may adversely affect the quality of resultant antigen-specific humoral responses, as seen in the case of RBD-p(Man-TLR7) (Fig. S4). This is because intracellular processing in the endosomes of APCs is required for release of the antigen from the rest of the construct, whereas humoral responses are partly dependent on extracellular interactions with B cell receptors in the germinal centers of secondary lymphoid organs. For most antigens, however, this will not be a relevant issue, and it may be possible to optimize the polymer to protein ratio if this issue does arise. In conclusion, to address a global need for next-generation vaccines against SARS-CoV-2, we have developed the Spike-p(Man-TLR7) vaccine platform and demonstrated its efficacy in mice. We found that conjugating the Spike protein to our polymeric glyco-adjuvant improves Spike’s immunogenicity through inducing both potent neutra...
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.
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