Immunopharmacological evaluation of adjuvant efficacy of Monophosphoryl lipid-A and CpG ODN with SARS-CoV-2 RBD antigen

  1. Shainy Sambyal
  2. Vemireddy Sravanthi
  3. Ritika Khatri
  4. Gazala Siddqui
  5. Shubbir Ahmed
  6. Sweety Samal
  7. Halmuthur M Sampath Kumar

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Abstract

SARS-CoV-2 infection has made the mankind to witness most sever and serious pandemic situation in the history. Millions of people have suffered and are still suffering with this infection which has caused a mass mortality in the past three years. Development of an effective vaccine to control the spread of infection and to prevent this viral infection is need of the hour. Adjuvanted vaccines have proven their efficacy in controlling many other viral infections like flu, keeping this context in view we have evaluated the immunopharmacological efficacy of two adjuvants MPL-A and CpG ODN in combination with MF59 emulsion against SARS-CoV-2 antigen. From the data obtained we can infer that both the adjuvants were capable of eliciting a potent antibody response against antigen alone and MF59 groups. Comparatively MPL-A was eliciting a Th1 polarized response in terms of IgG2a and cytokine production. Both the adjuvants were capable of enhancing the CD 4, 8 and 19 cell populations. Overall the pre clinical evaluation has given a clue of the effectiveness of MPL-A and CpG adjuvants against SARS-CoV-2 antigen.

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  1. ScreenIT

    SciScore for 10.1101/2022.04.04.486920: (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.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    flurochrome conjugated FACS anti mouse antibodies-CD16/32, CD4, CD8, CD19 (BD, USA), TMB substrate (BD, USA), HRP streptavidin (BioLegend, Switzerland), HRP Goat anti-mouse IgG antibody (BioLegend, Switzerland)
    anti mouse antibodies-CD16/32 , CD4
    suggested: None
    CD8
    suggested: (BioLegend Cat# 391503, RRID:AB_2721611)
    CD19
    suggested: None
    anti-mouse IgG
    suggested: None
    The sera was stored at 0 C and later was used for antibody titer: IgG, IgG1, IgG2a. 2.2.3 Spleen and splenocyte isolation: Mice were sacrificed with light ether anesthesia.
    IgG
    suggested: None
    IgG1
    suggested: None
    IgG2a . 2.2.3 Spleen
    suggested: None
    The cells were then stained with 5μl of PE-antiCD4, FITC-A antiCD8 and FITC-A antiCD19 flurochrome tagged antibodies for 45 min at 4°C.
    PE-antiCD4
    suggested: None
    FITC-A
    suggested: None
    antiCD8
    suggested: None
    antiCD19
    suggested: None
    Briefly, IFN-gamma, TNF-alpha and IL-6 coating antibody was diluted in bicarbonate-carbonate buffer and coated on ELISA plate for overnight at 4°C.
    IFN-gamma , TNF-alpha
    suggested: None
    IL-6
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    2.2 Methodology: 2.2.1 Immunization: 4-6 weeks female BALB/c mice purchased from RCC Laboratories India Private Limited were grouped as antigen alone (1μg/dose), MF59 either in combination with antigen alone or in combination with MPL-A and CpG ODN at varying concentration of 10, 20 and 30μg.
    BALB/c
    suggested: None
    Software and Algorithms
    SentencesResources
    The pellets so collected were resuspended in sheath fluid and cell population was quantified using BD FACSsuit software [8].
    BD FACSsuit
    suggested: None

    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

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  2. Version published to 10.1101/2022.04.04.486920v1 on bioRxiv