In contrast to T H 2-biased approaches, T H 1 COVID-19 vaccines protect Syrian hamsters from severe disease in the absence of dexamethasone-treatable vaccine-associated enhanced respiratory pathology

  1. Aileen Ebenig
  2. Samada Muraleedharan
  3. Julia Kazmierski
  4. Daniel Todt
  5. Arne Auste
  6. Martina Anzaghe
  7. André Gömer
  8. Dylan Postmus
  9. Patricia Gogesch
  10. Marc Niles
  11. Roland Plesker
  12. Csaba Miskey
  13. Michelle Gellhorn Serra
  14. Angele Breithaupt
  15. Cindy Hörner
  16. Carina Kruip
  17. Rosina Ehmann
  18. Zoltan Ivics
  19. Zoe Waibler
  20. Stephanie Pfaender
  21. Emanuel Wyler
  22. Markus Landthaler
  23. Alexandra Kupke
  24. Geraldine Nouailles
  25. Christine Goffinet
  26. Richard J.P. Brown
  27. Michael D. Mühlebach

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Abstract

Since December 2019, the novel human coronavirus SARS-CoV-2 has spread globally, causing millions of deaths. Unprecedented efforts have enabled development and authorization of a range of vaccines, which reduce transmission rates and confer protection against the associated disease COVID-19. These vaccines are conceptually diverse, including e.g. classical adjuvanted whole-inactivated virus, viral vectors, and mRNA vaccines.

We have analysed two prototypic model vaccines, the strongly T H 1-biased measles vaccine-derived candidate MeV vac2 -SARS2-S(H) and a T H 2-biased Alum-adjuvanted, non-stabilized Spike (S) protein side-by-side, for their ability to protect Syrian hamsters upon challenge with a low-passage SARS-CoV-2 patient isolate. As expected, the MeV vac2 -SARS2-S(H) vaccine protected the hamsters safely from severe disease. In contrast, the protein vaccine induced vaccine-associated enhanced respiratory disease (VAERD) with massive infiltration of eosinophils into the lungs. Global RNA-Seq analysis of hamster lungs revealed reduced viral RNA and less host dysregulation in MeV vac2 -SARS2-S(H) vaccinated animals, while S protein vaccination triggered enhanced host gene dysregulation compared to unvaccinated control animals. Of note, mRNAs encoding the major eosinophil attractant CCL-11, the T H 2 response-driving cytokine IL-19, as well as T H 2-cytokines IL-4, IL-5, and IL-13 were exclusively up-regulated in the lungs of S protein vaccinated animals, consistent with previously described VAERD induced by RSV vaccine candidates. IL-4, IL-5, and IL-13 were also up-regulated in S-specific splenocytes after protein vaccination. Using scRNA-Seq, T cells and innate lymphoid cells were identified as the source of these cytokines, while Ccl11 and Il19 mRNAs were expressed in lung macrophages displaying an activated phenotype. Interestingly, the amount of viral reads in this macrophage population correlated with the abundance of Fc-receptor reads. These findings suggest that VAERD is triggered by induction of T H 2-type helper cells secreting IL-4, IL-5, and IL-13, together with stimulation of macrophage subsets dependent on non-neutralizing antibodies. Via this mechanism, uncontrolled eosinophil recruitment to the infected tissue occurs, a hallmark of VAERD immunopathogenesis. These effects could effectively be treated using dexamethasone and were not observed in animals vaccinated with MeV vac2 -SARS2-S(H).

Taken together, our data validate the potential of T H 2-biased COVID-19 vaccines and identify the transcriptional mediators that underlie VAERD, but confirm safety of T H 1-biased vaccine concepts such as vector-based or mRNA vaccines. Dexamethasone, which is already in use for treatment of severe COVID-19, may alleviate such VAERD, but in-depth scrutiny of any next-generation protein-based vaccine candidates is required, prior and after their regulatory approval.

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    SciScore for 10.1101/2021.12.28.474359: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    RandomizationSix to 12-weeks old Syrian hamsters (Envigo RMS, Venray, Netherlands) were randomized for age- and sex-matched groups.
    BlindingH&E stained slices were subjected to histopathologic analyses on blinded samples.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Cells: Vero (African green monkey kidney; ATCC# CCL-81) and Vero clone E6 (ATCC# CRL-1586) cells were purchased from ATCC (Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Sigma Aldrich, Steinheim, Germany) supplemented with 10% Fetal bovine serum (FBS; Sigma Aldrich) and 2 mM L-glutamine (L-Gln, Sigma Aldrich).
    Vero clone E6
    suggested: None
    Stocks were generated by infection of Vero cells at an MOI = 0.03, and viruses in P3 or P4 were used for vaccination experiments.
    Vero
    suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
    SARS-CoV-2 isolate MUC-IMB1 (Böhmer et al., 2020) was used in passage 3 on Vero-E6 cells after isolation from the patient as described before (Hörner et al., 2020).
    Vero-E6
    suggested: None
    Subsequently, the virus-serum mixture was added to 1 x104 Vero or Vero E6 cells seeded 3 h before in 96 well plates (Thermo Fisher Scientific, Ulm, Germany).
    Vero E6
    suggested: None
    Software and Algorithms
    SentencesResources
    Cells: Vero (African green monkey kidney; ATCC# CCL-81) and Vero clone E6 (ATCC# CRL-1586) cells were purchased from ATCC (Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM, Sigma Aldrich, Steinheim, Germany) supplemented with 10% Fetal bovine serum (FBS; Sigma Aldrich) and 2 mM L-glutamine (L-Gln, Sigma Aldrich).
    FBS; Sigma Aldrich
    suggested: None
    Reactions were run in 96-well plates (Bio-Rad Laboratories, Hercules, CA) using CFX96 qPCR cycler (Bio-Rad Laboratories) and 5 μl RNA in a total reaction volume of 25 μl in triplicates.
    Bio-Rad Laboratories
    suggested: (Bio-Rad Laboratories, RRID:SCR_008426)
    Data were analysed using CellRanger v5.0 (10X Genomics) using hamster and SARS-CoV-2 genome scaffolds, and the R packages Seurat v4.0
    Seurat
    suggested: (SEURAT, RRID:SCR_007322)
    (Hao et al., 2021) and DoRothEA v3.12 (Holland et al., 2020) were used for cell clustering, annotation, and transcription factor activity analysis.
    DoRothEA
    suggested: (Scayle, RRID:SCR_019064)
    Gene set variation analysis (GSVA) was performed using the GSVA R package (Hänzelmann et al., 2013) and gene set enrichment analysis was performed using the clusterProfiler R package (Wu et al., 2021).
    clusterProfiler
    suggested: (clusterProfiler, RRID:SCR_016884)

    Results from OddPub: Thank you for sharing your code and data.

    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    As already pointed out by DiPiazza et al, drawing conclusions from animal models and extending these observations to the immunological situation in humans may be a difficult task (DiPiazza et al., 2021), considering the limitations of the system, especially in the absence of experience with vaccines targeting SARS-CoV and MERS-CoV and evidence for VAERD of all vaccine concepts including whole-inactivated viruses in humans. However, we were able to replicate VAERD induced by a protein based COVID-19 vaccine candidate with a very similar phenotype in a second animal model, Syrian hamsters, using an unmodified low-passage virus isolate. Therefore, our data strongly support the idea to monitor vaccinated human patients that experience a break-through infection closely. In any case, while our experimental vaccines mimic, but are not the same as the authorized vaccines, these and previously published data by diPiazza et al. point at considerably few concerns for vaccines developed to trigger TH1-biased responses such as viral vector platform-based vaccines and mRNA vaccines. Moreover, even if VAERD as observed in our model should occur in human patients, this immunopathology would be treatable by dexamethasone, which revealed to be an effective medication for severe courses of COVID-19, anyway (Tomazini et al., 2020; Horby et al., 2021). This would be good news also for putative VAERD being mistakenly diagnosed as a variant of the usual forms of severe COVID-19 in a naïve patient. ...

    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.

    About SciScore

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  2. Version published to 10.1101/2021.12.28.474359v1 on bioRxiv