An effective, safe and cost-effective cell-based chimeric vaccine against SARS-CoV2

  1. Zhenguo Cheng
  2. Danhua Zhang
  3. XiaoWen Liu
  4. Jinxin Miao
  5. Jianyao Wang
  6. Haoran Guo
  7. Wenli Yan
  8. Zhe Zhang
  9. Na Zhang
  10. Jingjing Wang
  11. Shuangshuang Lu
  12. Zhongxian Zhang
  13. Wei Liu
  14. Hong Liu
  15. Yi Zhang
  16. Lirong Zhang
  17. Jianzeng Dong
  18. Nicholas R Lemoine
  19. Yaohe Wang

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Abstract

More than one hundred vaccines against SARS-CoV-2 have been developed and some of them have entered clinical trials, but the latest results revealed that these vaccines still face great challenges. Here, we developed a novel cell-based gp96-Ig-secreting chimeric vaccine which is composed of two viral antigens, the RBD of spike protein, and a truncated nucleocapsid protein that could induce epitope-specific cytotoxic T lymphocytes but low antibody response. Syrian hamsters immunized with the cell-based vaccine produced high level of SARS-CoV-2 specific NAbs and specific T cell immunity which could eliminate RBD-truncated N-expressing cells, without the induction of antibody against N protein and other observed toxicity. This study provides a proof of concept for clinical testing of this safe, effective and cost-effective vaccine against SARS-CoV2 infection.

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  1. ScreenIT

    SciScore for 10.1101/2020.08.19.258244: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementConsent: Serum samples from recovered COVID-19 patients: 6 serum samples from recovered COVID-19 patients were collected in the First Affiliated Hospital of Zhengzhou University with the patients’ written consent and was approved by the Ethics Committee of Zhengzhou University.
    IACUC: All animals used in this study were maintained under specific pathogen-free conditions in Laboratory Animal Center of Zhengzhou University, and treatments was in accordance with the NIH Animal Care and Use Committee regulations.
    RandomizationAnimals: Female,12-week-old Syrian hamster were purchased from Vitalriver (Beijing), and randomly divided into 4 groups (n=9).
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Antibodies used in this study were listed as the following, rabbit anti-RBD PAb (polyclonal antibody) (Sino Biological, #40592-T62), rabbit anti-Nucleocapsid PAb (Sino Biological, #40588-T62), mouse anti-GAPDH mAb (monoclonal antibody) (ProteinTech, 60004-1-Ig)
    anti-RBD
    suggested: None
    anti-Nucleocapsid
    suggested: None
    anti-GAPDH
    suggested: None
    Then cells were washed twice with cold PBS and stained with mouse anti-human HLA-A2-FITC antibody (Abcam) for 30 min on ice.
    anti-human HLA-A2-FITC
    suggested: (MBL International Cat# K0186-4, RRID:AB_592228)
    Experimental Models: Cell Lines
    SentencesResources
    Sequence of human ACE2 and secreting gp96 (without “KDEL” motif of C terminal) were amplified from Suit2 and 293T cell respectively.
    293T
    suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
    Cells: Human kidney cell line HEK-293T and hamster kidney cell line BHK21 were purchased from Cell Bank of Type Culture Collection Committee of Chinese Academy of Sciences.
    HEK-293T
    suggested: None
    BHK21
    suggested: ATCC Cat# CRL-6282, RRID:CVCL_1914)
    To generate BHK21-hACE2 or C-Vac overexpressing cells, 5×104 cells (500 μL) BHK21 or 293T-gp96-hFc cells were seeded into 24-well plates, then 1 ml supernatant containing proper lentivirus and 5 μg/ml polybrene (Sigma) was added.
    293T-gp96-hFc
    suggested: None
    Group1-4 were vaccinated with 1×107 293T-gp96-hFc control cells, viable 293T-C-Vac cells, 293T-C-Vac cells treated with 5μg/ml mitomycin (MCE) for 4h and freeze-thaw treated 293T-C-Vac cells lysates, respectively.
    293T-C-Vac
    suggested: None
    Software and Algorithms
    SentencesResources
    Subsequently, the fluorescence intensity of cells were analyzed using a BD FACSAria (BD Biosciences Immunocytometry Systems) after washed twice with cold PBS.
    BD Biosciences Immunocytometry Systems
    suggested: None
    Statistics analysis: All statistical tests were performed by Graphpad Prism 7 using Student’s t-test (unpaired, two-tailed).
    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.

    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.

  2. Version published to 10.1101/2020.08.19.258244v1 on bioRxiv