Humoral cross-reactivity towards SARS-CoV-2 in young children with acute respiratory infection with low-pathogenicity coronaviruses

  1. Nitin Dhochak
  2. Tanvi Agrawal
  3. Heena Shaman
  4. Naseem Ahmed Khan
  5. Prawin Kumar
  6. Sushil K. Kabra
  7. Guruprasad R. Medigeshi
  8. Rakesh Lodha

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Abstract

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  1. Version published to 10.1016/j.jcvp.2022.100061
  2. ScreenIT

    SciScore for 10.1101/2021.10.01.21264349: (What is this?)

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

    Table 1: Rigor

    EthicsIRB: The current study was conducted after obtaining clearance from Institute Ethics Committee with waiver of individual consent [IECPG 756/23.12.2020; RT-01/2020] Virus microneutralization assay: Vero E6 cells (NCCS, Pune, India) were maintained in DMEM high glucose medium (HiMedia) supplemented with 10% heat-inactivated FBS, 100U of penicillin and 100 µg of streptomycin in 5% CO2 incubator.
    Consent: The current study was conducted after obtaining clearance from Institute Ethics Committee with waiver of individual consent [IECPG 756/23.12.2020; RT-01/2020] Virus microneutralization assay: Vero E6 cells (NCCS, Pune, India) were maintained in DMEM high glucose medium (HiMedia) supplemented with 10% heat-inactivated FBS, 100U of penicillin and 100 µg of streptomycin in 5% CO2 incubator.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line AuthenticationAuthentication: The convalescent serum samples from children infected with HCoVs OC43, NL63 and 229E were evaluated for antibodies against respective human coronaviruses by immunofluorescence assay.

    Table 2: Resources

    Antibodies
    SentencesResources
    Cells were then stained with anti-spike RBD antibody at 1:4000 dilution (Sino Biologicals; 40592-T62) for 1 h, followed by HRP-conjugated anti-rabbit antibody at 1:4000 dilution (Invitrogen; G-21234) for 1 h.
    anti-spike RBD
    suggested: None
    anti-rabbit
    suggested: (Innovative Research Cat# G-21234, RRID:AB_1500696)
    The secondary antibodies used were goat anti-human IgG-Alexa 488 (A11013 - ThermoFisher) for human sera or goat anti-rabbit IgG-Alexa 488 (A21206-ThermoFisher) for nucleocapsid antibodies.
    anti-human IgG-Alexa
    suggested: None
    anti-rabbit IgG-Alexa
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    The current study was conducted after obtaining clearance from Institute Ethics Committee with waiver of individual consent [IECPG 756/23.12.2020; RT-01/2020] Virus microneutralization assay: Vero E6 cells (NCCS, Pune, India) were maintained in DMEM high glucose medium (HiMedia) supplemented with 10% heat-inactivated FBS, 100U of penicillin and 100 µg of streptomycin in 5% CO2 incubator.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Human hepatoma cells (Huh-7) (Japanese Collection of Research Bioresources Cell Bank) were grown at 37° C in Dulbecco’s minimum essential medium (DMEM) (Lonza 12-707F) containing above additives and non-essential amino acids (Gibco-111400-50).
    Huh-7
    suggested: None
    Virus propagation: HCoV-OC43 was propagated in BHK-21 and HCoV-NL63 in LLC MK2 cells in MEM supplemented with 2% heat-inactivated FBS, 100 U/mL of penicillin, and 100 µg/L of streptomycin at 34° C under a humidified atmosphere of 5% CO2 for 4 days and 6 days respectively.
    BHK-21
    suggested: None
    Virus dilutions of 1:10 for HCoV-NL63 and HCoV-OC43 infection in LLC MK2 and 1:10000 for HCoV-229E infection in Huh-7 cells were determined as optimum for infection without cytopathic effect.
    HCoV-NL63
    suggested: RRID:CVCL_RW88)
    Software and Algorithms
    SentencesResources
    The microspots were quantified by AID iSPOT reader (ELR08IFL; AID GmbH, Strassberg, Germany) using AID EliSpot 8.0 software. 50% neutralization values were calculated with four-parameter logistic regression using GraphPad Prism 7.0e software.
    GraphPad Prism
    suggested: (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: 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

    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 10.1101/2021.10.01.21264349 on medRxiv