SARS‐CoV‐2 infection as a trigger of humoral response against apolipoprotein A‐1

  1. Sabrina Pagano
  2. Sabine Yerly
  3. Benjamin Meyer
  4. Catherine Juillard
  5. Noémie Suh
  6. Christophe Le Terrier
  7. Jean‐Pierre Daguer
  8. Lluc Farrera‐Soler
  9. Sofia Barluenga
  10. Giovanni Piumatti
  11. Oliver Hartley
  12. Barbara Lemaitre
  13. Christiane S. Eberhardt
  14. Claire‐Anne Siegrist
  15. Isabella Eckerle
  16. Silvia Stringhini
  17. Idris Guessous
  18. Laurent Kaiser
  19. Jerome Pugin
  20. Nicolas Winssinger
  21. Nicolas Vuilleumier

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Abstract

Background

Unravelling autoimmune targets triggered by SARS‐CoV‐2 infection may provide crucial insights into the physiopathology of the disease and foster the development of potential therapeutic candidate targets and prognostic tools. We aimed at determining (a) the association between anti‐SARS‐CoV‐2 and anti‐apoA‐1 humoral response and (b) the degree of linear homology between SARS‐CoV‐2, apoA‐1 and Toll‐like receptor 2 (TLR2) epitopes.

Design

Bioinformatics modelling coupled with mimic peptides engineering and competition experiments were used to assess epitopes sequence homologies. Anti‐SARS‐CoV‐2 and anti‐apoA‐1 IgG as well as cytokines were assessed by immunoassays on a case‐control (n = 101), an intensive care unit (ICU; n = 126) and a general population cohort (n = 663) with available samples in the pre and post‐pandemic period.

Results

Using bioinformatics modelling, linear sequence homologies between apoA‐1, TLR2 and Spike epitopes were identified but without experimental evidence of cross‐reactivity. Overall, anti‐apoA‐1 IgG levels were higher in COVID‐19 patients or anti‐SARS‐CoV‐2 seropositive individuals than in healthy donors or anti‐SARS‐CoV‐2 seronegative individuals ( P  < .0001). Significant and similar associations were noted between anti‐apoA‐1, anti‐SARS‐CoV‐2 IgG, cytokines and lipid profile. In ICU patients, anti‐SARS‐CoV‐2 and anti‐apoA‐1 seroconversion rates displayed similar 7‐day kinetics, reaching 82% for anti‐apoA‐1 seropositivity. In the general population, SARS‐CoV‐2‐exposed individuals displayed higher anti‐apoA‐1 IgG seropositivity rates than nonexposed ones (34% vs 16.8%; P  = .004).

Conclusion

COVID‐19 induces a marked humoral response against the major protein of high‐density lipoproteins. As a correlate of poorer prognosis in other clinical settings, such autoimmunity signatures may relate to long‐term COVID‐19 prognosis assessment and warrant further scrutiny in the current COVID‐19 pandemic.

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  1. Version published to 10.1111/eci.13661
  2. Version published to 10.1101/2021.02.12.21251298v2 on medRxiv
  3. ScreenIT

    SciScore for 10.1101/2021.02.12.21251298: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Assessment of total antibodies against N antigen of SARS-CoV-2: Total antibodies against the N antigen of SARS-CoV-2 were measured on a Cobas e801 analyzer (Roche Diagnostics, Rotkreuz, Switzerland) according to the manufacturer’s instructions.
    antibodies against N
    suggested: None
    After six washing cycles, a 50 µL/well of signal antibody (alkaline phosphatase-conjugated anti-human IgG; Sigma-Aldrich, St Louis, MO, USA, ref: A-3150), diluted 1:1000 in a PBS/BSA 2% solution, was added and incubated for 1 h at 37 C.
    anti-human IgG
    suggested: (Sigma-Aldrich Cat# A3150, RRID:AB_258051)
    Commercial polyclonal anti-apoA-1 IgG (Academy Biomedical Company, Houston, TX, USA, ref: 11A-G2) and polyclonal anti-Spike IgG (Thermo Fisher Scientific, Waltham, MA, USA, ref: PA5-81795) with their respective control antibodies: goat IgG (Meridian Life Science, Memphis, TN, USA, ref: A66200H) or rabbit IgG (Dako, Santa Clara, CA, USA, ref: X0903), were added to the coated plate at increasing concentrations (1 to 10 µg/ml) for 1 h.
    Commercial polyclonal anti-apoA-1 IgG
    suggested: None
    anti-apoA-1 IgG
    suggested: (BD Biosciences Cat# 612331, RRID:AB_399697)
    11A-G2
    suggested: (Academy Biomedical Company Cat# 11A-G2, RRID:AB_1238781)
    anti-Spike IgG
    suggested: None
    IgG
    suggested: None
    rabbit IgG
    suggested: None
    Anti-goat or anti-rabbit alkaline phosphatase-conjugated antibodies (
    Anti-goat or anti-rabbit alkaline phosphatase-conjugated antibodies
    suggested: None
    anti-rabbit
    suggested: None
    Competition experiments were done using our standard anti-apoA-1 IgG protocol (21–27), which involved coating the plate with apo-A-1 or Spike protein, then increasing concentrations of polyclonal anti-Spike antibodies or polyclonal anti-apoA-1 IgG (5 to 40 µg/ml) were added to the plate for 30 min, and then pooled sera from seven anti-apoA-1 IgG were subsequently incubated onto the plate for one hour at 37°C.
    anti-Spike
    suggested: None
    Software and Algorithms
    SentencesResources
    Sequence homology analyses, Spike-apoA-1 and Spike-TLR2 mimic peptides synthesis: Homologies between Spike and Nucleocapsid epitopes (13–16), TLR2 or c-ter apoA-1 were assessed using Clustal Omega and BlastP sequence alignment.
    Clustal Omega
    suggested: (Clustal Omega, RRID:SCR_001591)
    BlastP
    suggested: (BLASTP, RRID:SCR_001010)
    All analyses were performed using Statistica software (version 13.5.0.17,
    Statistica
    suggested: (STATISTICA , RRID:SCR_014213)

    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:
    We acknowledge several limitations of the present work. Firstly, our analysis used linear epitope mapping and doesn’t consider potential conformational epitopes which may further contribute to the association between the anti-apoA-1 and anti-N serology. Nevertheless, as those two linear immunodominant epitopes are exposed to the immune system regardless of the conformation of spike (monomeric or trimeric) (13, 14, 15), potential conformational issues are unlikely to have blunted the present conclusions. Secondly, the relatively limited number of patients enrolled in this study impeded our ability to complete the survival analyses with the desired level of details regarding the prognostic value of anti-Spike/TLR2, which needs replication in larger prospective cohorts. Thirdly, due to the fact that our institution became a fully COVID-19 dedicated infrastructure, we could not identify non-COVID-19 matched controls during the studies inclusion period. Fourthly, because the aa579-587 sequence of Spike shared sequence homology with the aa 456-464 of TLR2 corresponding to the PAMP/DAMP sensing of TLRs, knowing whether direct anti-S1 IgG-mediated TLR2 stimulation could per se elicit a pro-inflammatory response remains to be explored. Fourth, as anti-apoA-1 IgG are not to be predominantly ascribed to CV prognosis, knowing whether the SARS-CoV-2-induced anti-apoA-1 IgG seroconversion could modulate the long-term CV prognosis of COVID-19 patients remains elusive. Furthermore, being pri...

    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.

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  4. Version published to 10.1101/2021.02.12.21251298v1 on medRxiv