A recombinant protein containing influenza viral conserved epitopes and superantigen induces broad-spectrum protection

  1. Yansheng Li
  2. Mingkai Xu
  3. Yongqiang Li
  4. Wu Gu
  5. Gulinare Halimu
  6. Yuqi Li
  7. Zhichun Zhang
  8. Libao Zhou
  9. Hui Liao
  10. Songyuan Yao
  11. Huiwen Zhang
  12. Chenggang Zhang

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Abstract

Influenza pandemics pose public health threats annually for lacking vaccine that provides cross-protection against novel and emerging influenza viruses. Combining conserved antigens that induce cross-protective antibody responses with epitopes that activate cross-protective T cell responses might be an attractive strategy for developing a universal vaccine. In this study, we constructed a recombinant protein named NMHC that consists of influenza viral conserved epitopes and a superantigen fragment. NMHC promoted the maturation of bone marrow-derived dendritic cells and induced CD4 + T cells to differentiate into Th1, Th2, and Th17 subtypes. Mice vaccinated with NMHC produced high levels of immunoglobulins that cross-bound to HA fragments from six influenza virus subtypes with high antibody titers. Anti-NMHC serum showed potent hemagglutinin inhibition effects to highly divergent group 1 (H1 subtype) and group 2 (H3 subtype) influenza virus strains. Furthermore, purified anti-NMHC antibodies bound to multiple HAs with high affinities. NMHC vaccination effectively protected mice from infection and lung damage when exposed to two subtypes of H1N1 influenza virus. Moreover, NMHC vaccination elicited CD4 + and CD8 + T cell responses that cleared the virus from infected tissues and prevented virus spread. In conclusion, this study provides proof of concept that NMHC vaccination triggers B and T cell immune responses against multiple influenza virus infections. Therefore, NMHC might be a candidate universal broad-spectrum vaccine for the prevention and treatment of multiple influenza viruses.

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  1. Version published to 10.7554/elife.71725 on eLife
  2. ScreenIT

    SciScore for 10.1101/2021.07.08.451596: (What is this?)

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

    Table 1: Rigor

    EthicsIACUC: All animal procedures were performed in accordance with Institutional Animal Care and Use Committee (IACUC) guidelines and have been approved by the IACUC of University of Chinese Academy of Sciences. Reagents: The expression vector pET-28a-sec2 containing full-length cDNA of SEC2 was constructed previously and preserved in our laboratory(Fu et al., 2017).
    Field Sample Permit: Funding: This work was supported by Strategic Priority Research Program of the Chinese Academy of Sciences Grant (XDA12020225),
    Sex as a biological variableAnimals: Female BALB/c mice (4–6weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co. Ltd (Beijing, China) and maintained under specific pathogen-free conditions.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Monoclonal CD3-FITC/APC, CD11c-APC, CD80-PE, CD86-FITC, MHC II-FITC, IL-4-FITC, IL-10-FITC, IL-17-PE, and IFN-γ-APC Antibodies (Abs) were purchased from BioLegend (San Diego, CA).
    CD80-PE
    suggested: None
    CD86-FITC
    suggested: None
    IL-17-PE
    suggested: None
    The maturation of BMDCs were evaluated through detecting the cell surface markers including CD80, CD86, and MHC II with fluorescent labeled antibodies, respectively, followed by analyzing in a FACScalibur flow cytometer.
    CD80
    suggested: None
    CD86
    suggested: None
    Then cells were intracellular stained with fluorescent labeled antibodies against IL-4 (for Th1 subtype), IFN-γ (for Th2 subtype), IL-17A (for Th17 subtype), and IL-10 (for Th10 subtype), respectively.
    IL-4
    suggested: None
    Th1 subtype) , IFN-γ ( for Th2 subtype)
    suggested: None
    Th17 subtype) ,
    suggested: None
    IL-10
    suggested: None
    Influenza-specific IgG antibody secreting cells (ASCs) were enumerated with ELISPOT.
    Influenza-specific IgG
    suggested: None
    This folder contains the overlays of binding kinetics of antibodies to HAs (HA of H1, H2, H3 and H5) at different concentrations were detected with Biacore (individual files are named “the binding kinetics of antibodies to H1.txt”, “the binding kinetics of antibodies to H2.txt”, “the binding kinetics of antibodies to H3.txt”,
    H2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    After incubation for 1 h at 37°C, 1.5×104 MDCK cells supplemented with 2 μg/mL trypsin and 0.5% Bovine Serum Albumin (BSA) in DMEM media were added to each well.
    MDCK
    suggested: CLS Cat# 602280/p823_MDCK_(NBL-2, RRID:CVCL_0422)
    Experimental Models: Organisms/Strains
    SentencesResources
    Animals: Female BALB/c mice (4–6weeks old) were purchased from Beijing Vital River Laboratory Animal Technology Co. Ltd (Beijing, China) and maintained under specific pathogen-free conditions.
    BALB/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Recombinant DNA
    SentencesResources
    All animal procedures were performed in accordance with Institutional Animal Care and Use Committee (IACUC) guidelines and have been approved by the IACUC of University of Chinese Academy of Sciences. Reagents: The expression vector pET-28a-sec2 containing full-length cDNA of SEC2 was constructed previously and preserved in our laboratory(Fu et al., 2017).
    pET-28a-sec2
    suggested: None
    The encoding DNA fragment were ligated into the expression vectors pET-28a (+) plasmid and transformed into E. coli BL21(DE3), and the positive clones were verified by DNA sequencing.
    pET-28a ( + )
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical analysis: The data were analyzed by Student’s t-test, one-way analysis of variance (ANOVA) and followed by a suitable post hoc test using the SPSS 26.0 and GraphPad Prism Software (version 6.0c).
    SPSS
    suggested: (SPSS, RRID:SCR_002865)
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Funding: This work was supported by Strategic Priority Research Program of the Chinese Academy of Sciences Grant (XDA12020225),
    Strategic Priority Research Program
    suggested: None
    Liaoning Revitalization Talents Program (XLYC1807226), Science and Technology Plan Projects of Shenyang City Grants (Z17-7-013), and Shenyang High-level Innovative Talents Program (RC190060)
    Revitalization Talents Program
    suggested: None
    Innovative Talents Program
    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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 11. 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.

    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 10.1101/2021.07.08.451596v1 on bioRxiv