A spike-ferritin nanoparticle vaccine induces robust innate immune activity and drives polyfunctional SARS-CoV-2-specific T cells
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Abstract
Potent cellular responses to viral infections are pivotal for long -lived protection. Evidence is growing that these responses are critical in SARS -CoV-2 immunity. Assessment of a SARS -CoV-2 spike ferritin nanoparticle (SpFN) immunogen paired with two distinct adjuvants, Alhydrogel® (AH) or Army Liposome Formulation containing QS-21 (ALFQ) demonstrated unique vaccine evoked immune signatures. SpFN+ALFQ enhanced recruitment of highly activated classical and non -classical antigen presenting cells (APCs) to the vaccine-draining lymph nodes of mice. The multifaceted APC response of SpFN+ALFQ vaccinated mice was associated with an increased frequency of polyfunctional spike -specific T cells with a bias towards T H 1 responses and more robust SARS-CoV-2 spike-specific recall response. In addition, SpFN+ALFQ induced K b spike (539-546) -specific memory CD8 + T cells with effective cytolytic function and distribution to the lungs. This epitope is also present in SARS-CoV, thus suggesting that generation of cross-reactive T cells may provide protection against other coronavirus strains. Our study reveals that a nanoparticle vaccine, combined with a potent adjuvant, generates effective SARS-CoV-2 specific innate and adaptive immune T cell responses that are key components to inducing long-lived immunity.
One Sentence Summary
SpFN vaccine generates multifactorial cellular immune responses.
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SciScore for 10.1101/2021.04.28.441763: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: The protocols were approved by the Institutional Animal Care and Use Committee at the Walter Reed Army Institute of Research [Assurance number D16-00596 (A4117-01)]. Sex as a biological variable Mice vaccination and tissue processing: Female C57BL/6 mice (5-6 weeks of age) were obtained from The Jackson Laboratory. Randomization Mice were age- and sex-matched and were randomly assigned to study groups. Blinding The APC flow cytometry and ELISpot analysis were performed by the investigators in a blinded fashion as they were not aware of which treatment group belonged to which adjuvant formulation. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sente… SciScore for 10.1101/2021.04.28.441763: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: The protocols were approved by the Institutional Animal Care and Use Committee at the Walter Reed Army Institute of Research [Assurance number D16-00596 (A4117-01)]. Sex as a biological variable Mice vaccination and tissue processing: Female C57BL/6 mice (5-6 weeks of age) were obtained from The Jackson Laboratory. Randomization Mice were age- and sex-matched and were randomly assigned to study groups. Blinding The APC flow cytometry and ELISpot analysis were performed by the investigators in a blinded fashion as they were not aware of which treatment group belonged to which adjuvant formulation. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After the incubation period, cells were stained with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Invitrogen), followed by surface staining with antibodies specific for the following cell surface markers (BUV737 anti -CD3, BUV395 anti-CD4, BV711 anti-CD8 (table S4), obtained from either BD Biosciences, Thermo Fisher Scientific or Biolegend, washed twice with FACS buffer and then fixed/permeabilized for 40 mins at 4°C in the dark using the eBioscience™ Intracellular Fixation & Permeabilization Buffer Set (Thermo Fisher Scientific) as per the manufacturer’s instructions. anti -CD3, BUV395suggested: (BD Biosciences Cat# 563878, RRID:AB_2631259)anti-CD4suggested: NoneBV711suggested: Noneanti-CD8suggested: NoneCells were then incubated with a panel of intracellular antibodies specific for the following cytokines (V450 anti -IFN-γ, FITC anti-TNF-α, PerCP-Cy5 anti-IL-4, PE anti IL-2; table S4) for 30 min at 4°C, washed twice, and resuspended in FACS buffer. anti -IFN-γ, FITCsuggested: Noneanti-IL-4suggested: Noneanti IL-2; table S4suggested: NoneELISpot: Multiscreen plates (Millipore, Bedford MA) were coated with anti-IFN-γ antibody (capture mAb; 1 µg/mL) according to the manufacturer’s instructions (R&D Systems, Minneapolis, MN). anti-IFN-γsuggested: NonePlates were incubated with biotinylated anti -IFN-γ antibody (detecting mAb; 1 µg/mL) overnight at 4°C. anti -IFN-γsuggested: NoneIn the last 20 min of tetramer incubation, the following mix of fluorescent antibodies were added: BUV737-anti-mouse CD3, BUV395-anti-mouse CD4, BV711-anti-mouse CD8a and antibodies directed towards intracellular cytokines such as V450 anti -IFN-γ, FITC anti-TNF-α, and APC anti IL-2 (Supplementary Table 4), then washed twice and resuspended in FACS buffer. CD3suggested: NoneBUV395-anti-mouse CD4suggested: NoneBV711-anti-mousesuggested: NoneCD8asuggested: (BD Biosciences Cat# 560471, RRID:AB_1645282)Experimental Models: Cell Lines Sentences Resources The protein was transiently expressed as a soluble recombinant protein in mammalian Expi293 cells (Thermo Fisher Scientific). Expi293suggested: RRID:CVCL_D615)Experimental Models: Organisms/Strains Sentences Resources Mice vaccination and tissue processing: Female C57BL/6 mice (5-6 weeks of age) were obtained from The Jackson Laboratory. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources For vaccine preparations adjuvanted with ALFQ, SpFN (600 µg/mL) was mixed with ALFQ (1.5X) in a 1:2 volume ratio. ALFQsuggested: (aLFQ, RRID:SCR_005925)Evaluation of co-expression of different cytokines was performed using the FlowJo Boolean gate platform. FlowJosuggested: (FlowJo, RRID:SCR_008520)CFSE high and low solutions were made at 0.5 and 0.05 µM respectively using ThermoFisher CellTrace™ CFSE Cell Proliferation Kit (Catalog C34554). ThermoFisher CellTrace™suggested: NoneStatistical analyses were conducted using GraphPad Prism v.8.4.0 software. GraphPad Prismsuggested: (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: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT03186781 Completed Influenza HA Ferritin Vaccine, Alone or in Prime-Boost Regim… NCT03814720 Completed Dose, Safety, Tolerability and Immunogenicity of an Influenz… NCT04645147 Recruiting Safety and Immunogenicity of an Epstein-Barr Virus (EBV) gp3… NCT04579250 Recruiting Dose, Safety, Tolerability and Immunogenicity of an Influenz… NCT04784767 Recruiting SARS-COV-2-Spike-Ferritin-Nanoparticle (SpFN) Vaccine With A… 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 45. 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.
- Thank you for including a protocol registration statement.
Results from scite Reference Check: We found no unreliable references.
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