Recurrence of COVID-19 associated with reduced T-cell responses in a monozygotic twin pair

  1. Mateus V. de Castro
  2. Keity S. Santos
  3. Juliana S. Apostolico
  4. Edgar R. Fernandes
  5. Rafael R. Almeida
  6. Gabriel Levin
  7. Jhosiene Y. Magawa
  8. João Paulo S. Nunes
  9. Mirian Bruni
  10. Marcio M. Yamamoto
  11. Ariane C. Lima
  12. Monize V. R. Silva
  13. Larissa R. B. Matos
  14. Vivian R. Coria
  15. Erick C. Castelli
  16. Marilia O. Scliar
  17. Andreia Kuramoto
  18. Fernanda R. Bruno
  19. Lucas C. Jacintho
  20. Kelly Nunes
  21. Jaqueline Y. T. Wang
  22. Veronica P. Coelho
  23. Miguel Mitne Neto
  24. Rui M. B. Maciel
  25. Michel S. Naslavsky
  26. Maria Rita Passos-Bueno
  27. Silvia B. Boscardin
  28. Daniela S. Rosa
  29. Jorge Kalil
  30. Mayana Zatz
  31. Edecio Cunha-Neto

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Abstract

Recurrence of COVID-19 in recovered patients has been increasingly reported. However, the immune mechanisms behind the recurrence have not been thoroughly investigated. The presence of neutralizing antibodies (nAbs) in recurrence/reinfection cases suggests that other types of immune response are involved in protection against recurrence. Here, we investigated the innate type I/III interferon (IFN) response, binding and nAb assays and T-cell responses to severe acute respiratory distress syndrome coronavirus 2 (SARS-CoV-2) with IFN gamma (IFN γ ) enzyme-linked spot assay (ELISPOT) in three pairs of young adult monozygotic (MZ) twins with previous confirmed COVID-19, one of them presenting a severe recurrence four months after the initial infection. Twin studies have been of paramount importance to comprehend the immunogenetics of infectious diseases. Each MZ twin pair was previously exposed to SARS-CoV-2, as seen by clinical reports. The six individuals presented similar overall recovered immune responses except for the recurrence case, who presented a drastically reduced number of recognized SARS-CoV-2 T-cell epitopes on ELISPOT as compared to her twin sister and the other twin pairs. Our results suggest that the lack of a broad T-cell response to initial infection may have led to recurrence, emphasizing that an effective SARS-CoV-2-specific T-cell immune response is key for complete viral control and avoidance of clinical recurrence of COVID-19.

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  1. Version published to 10.1098/rsob.210240
  2. ScreenIT

    SciScore for 10.1101/2021.03.26.21253645: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethics Statement: The study was approved by the Committee for Ethics in Research of the Institute of Biosciences at the University of São Paulo (CAAE 34786620.2.0000.5464) and all participants (or their parents) provided a written consent.
    Consent: Ethics Statement: The study was approved by the Committee for Ethics in Research of the Institute of Biosciences at the University of São Paulo (CAAE 34786620.2.0000.5464) and all participants (or their parents) provided a written consent.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Cells were seeded at 105 cells/well in MultiScreen MAIPS Filter Plates (Merck) using coating and secondary anti-IFNγ antibodies (BD Biosciences).
    anti-IFNγ
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Enzyme-linked immunosorbent assay ELISA was performed using 96-well high-binding half-area polystyrene plates coated overnight at 4°C with 4 μg/mL of Spike protein, 2 μg/mL Nucleocapsid protein (NP) (Kindly provided by Dr Ricardo Gazzinelli, UFMG) or 0.8μg/mL of the RBD domain from human endemic coronaviruses HKU-1, OC43, NL63, and 229E, all expressed in HEK293T cells.
    HEK293T
    suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
    HT1080 expressing ACE2 cells (HT1080/ACE2) and plasmids HIV-1NLΔEnv-NanoLuc and pSARS-CoV-2-SΔ19 were kindly provided by Dr. Paul D.
    HT1080
    suggested: None
    Software and Algorithms
    SentencesResources
    50% inhibitory dilution (IC50) was calculated using Prism software (v7.0, GraphPad) after subtraction of the background RLUs in the control wells (cells only).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Sequencing data was analyzed following bwa-mem and GATK Best Practices workflow, quality control and annotation was performed as previously described (15).
    GATK
    suggested: (GATK, RRID:SCR_001876)

    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:
    Our study has limitations. We report observations for only one case of COVID-19 recurrence and cannot ensure that our findings are common in COVID-19 recurrence. We cannot conclude whether this recurrence event is a bona fide reinfection or a viral relapse in a chronic carrier since we did not genotype the SARS-CoV-2 virus. Most reinfection episodes were reported to be asymptomatic or mild (5). However, the 4-month interval between the two events in an exposed health care worker with a more severe clinical presentation is more suggestive of reinfection due to re-exposure than of a viral relapse. In short, our results suggest that the failure in inducing a broad T cell response might have enhanced susceptibility to COVID-19 recurrence in the reported case. Our data may support a prime role for T cells in protection against reinfection. Given the increased concern that SARS-CoV-2 variants escaping antibody neutralization could give rise to a massive raise in reinfection (24, 25), our case stresses the importance of T cell immune responses in protection against reinfection. This is in line with the reported lack of deleterious effect of virus variants in the cellular immune response (26). Further investigation in a larger cohort can shed light on whether T cell dysfunction is a common mechanism for recurrence of COVID-19.

    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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  3. Version published to 10.1101/2021.03.26.21253645 on medRxiv