Characterization of humoral and SARS-CoV-2 specific T cell responses in people living with HIV

  1. Aljawharah Alrubayyi
  2. Ester Gea-Mallorquí
  3. Emma Touizer
  4. Dan Hameiri-Bowen
  5. Jakub Kopycinski
  6. Bethany Charlton
  7. Natasha Fisher-Pearson
  8. Luke Muir
  9. Annachiara Rosa
  10. Chloe Roustan
  11. Christopher Earl
  12. Peter Cherepanov
  13. Pierre Pellegrino
  14. Laura Waters
  15. Fiona Burns
  16. Sabine Kinloch
  17. Tao Dong
  18. Lucy Dorrell
  19. Sarah Rowland-Jones
  20. Laura E. McCoy
  21. Dimitra Peppa

Reviewed by

Read the full article See related articles

Listed in

Log in to save this article

Abstract

There is an urgent need to understand the nature of immune responses against SARS-CoV-2, to inform risk-mitigation strategies for people living with HIV (PLWH). Here we show that the majority of PLWH with ART suppressed HIV viral load, mount a detectable adaptive immune response to SARS-CoV-2. Humoral and SARS-CoV-2-specific T cell responses are comparable between HIV-positive and negative subjects and persist 5-7 months following predominately mild COVID-19 disease. T cell responses against Spike, Membrane and Nucleoprotein are the most prominent, with SARS-CoV-2-specific CD4 T cells outnumbering CD8 T cells. We further show that the overall magnitude of SARS-CoV-2-specific T cell responses relates to the size of the naive CD4 T cell pool and the CD4:CD8 ratio in PLWH. These findings suggest that inadequate immune reconstitution on ART, could hinder immune responses to SARS-CoV-2 with implications for the individual management and vaccine effectiveness in PLWH.

Article activity feed

  1. Version published to 10.1038/s41467-021-26137-7
  2. ScreenIT

    SciScore for 10.1101/2021.02.15.431215: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: Ethics statement: The protocols for the human study were approved by the local Research Ethics Committee (REC) – Berkshire (REC 16/SC/0265).
    Consent: The study conformed to the Helsinki declaration principles and Good Clinical Practice (GCP) guidelines and all subjects enrolled into the study provided written informed consent.
    IACUC: All participants were recruited at the Mortimer Market Centre for Sexual Health and HIV Research and the Royal Free Hospital (London, UK) following written informed consent as part of a study approved by the local ethics board committee.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Briefly, ELISPOT plates (S5EJ044I10; Merck Millipore, Darmstadt, Germany) pre-wetted with 30 µl of 70% ethanol for a maximum of 2 minutes, washed with sterile PBS and coated overnight at 4 °C with anti–IFN-γ antibody (10 µg/ml in PBS; clone 1-D1K; Mabtech, Nacka Strand, Sweden).
    anti–IFN-γ
    suggested: None
    After overnight rest, PBMCs were stimulated for 6 h with 2 μg/mL of SARS-CoV-2 peptide pools, Influenza, HIV-1 Gag or cytomegalovirus (CMV)-pp65 peptide pools, or with 0.005% dimethyl sulphoxide (DMSO) as a negative control in the presence of αCD28/αCD49d co-Stim antibodies (1 μg ml−1) GolgiStop
    Influenza
    suggested: None
    HIV-1 Gag
    suggested: None
    cytomegalovirus ( CMV)-pp65
    suggested: None
    (containing Monensin, 2 μmol/L), GolgiPlug (containing brefeldin A, 10 μg ml−1) (BD Biosciences) and anti-CD107α BV421 antibody (BD Biosciences).
    Monensin , 2
    suggested: None
    anti-CD107α BV421
    suggested: None
    After stimulation, cells were washed and stained with anti-CCR7 (BioLegend) for 30 min at 37 °C and then surface stained at 4°C for 20 min with different combinations of surface antibodies in the presence of fixable live/dead stain (Invitrogen).
    anti-CCR7
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Pseudovirus production and neutralization assays: HIV-1 particles pseudotyped with SARS-Cov-2 spike were produced by seeding 3×106 HEK- 293T cells in 10ml complete DMEM (DMEM supplemented with 10% FBS, L-Glutamine, 100 IU/ml penicillin and 100 µg/ml streptomycin) in a T-75 culture flask.
    293T
    suggested: None
    HeLa ACE-2 cells (gift from James E Voss, Scripps Institute) were then added to the assay (10,000 cells per 100 μL per well).
    HeLa ACE-2
    suggested: None
    Software and Algorithms
    SentencesResources
    ODs were measured using a MultiskanFC (Thermofisher) plate reader at 405nm and S1 & N-specific IgG titers interpolated from the IgG standard curve using 4PL regression curve-fitting on GraphPad Prism 8.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    (BD Bioscience) and data analysed using FlowJo 10 (TreeStar).
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    : Prism 8 (GraphPad Software) was used for statistical analysis as follows: the Mann–Whitney U-test was used for single comparisons of independent groups, the Wilcoxon-test paired t-test was used to compare two paired groups.
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Polyfunctionality tests were performed in SPICE version 6.0.
    SPICE
    suggested: (SPICE, RRID:SCR_016603)

    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:
    There are limitations to this study. The observed heterogeneity in the magnitude of cellular and humoral responses that are not always fully coordinated highlights the need to consider additional putative factors as they relate to adaptive immunity. This cross-sectional study was not powered to study age and demographic differences according to the full spectrum of COVID-19 disease by HIV serostatus. Larger studies are required to determine the role of gender, racial and ethnicity effects, especially in areas of high HIV burden and additional comorbidities, to help identify individuals who are particularly vulnerable to the impact of SARS-CoV-2 infection and need targeted vaccination interventions. Nonetheless, the prospective, longitudinal design of this current study, integrating clinical parameters, antibody and T cell responses, will help address longer term protective immunity and emerging questions, such as immune responses to new SARS-CoV-2 variants (C Rees-Spear 2021; Houriiyah Tegally 2020; SA Kemp 2020; Sandile Cele 2021; Wibmer et al. 2021), and during the subsequent vaccination roll-out. Collectively, our results provide benchmark data into the facets of adaptive immunity against SARS-CoV-2 in the setting of treated HIV infection, providing evidence for medium-term durable antibody and cellular responses. Although reassuring, our data also have implications for PLWH with inadequate immune reconstitution, reflected in the low/inverted CD4:CD8 ratio, and potentially...

    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: Please consider improving the rainbow (“jet”) colormap(s) used on page 55. 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.
    • 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 10.1101/2021.02.15.431215v1 on bioRxiv