Zebrafish studies on the vaccine candidate to COVID-19, the Spike protein: Production of antibody and adverse reaction
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Abstract
Establishing new experimental animal models to assess the safety and immune response to the antigen used in the development of COVID-19 vaccine is an imperative issue. Based on the advantages of using zebrafish as a model in research, herein we suggest doing this to test the safety of the putative vaccine candidates and to study immune response against the virus. We produced a recombinant N-terminal fraction of the Spike SARS-CoV-2 protein and injected it into adult female zebrafish. The specimens generated humoral immunity and passed the antibodies to the eggs. However, they presented adverse reactions and inflammatory responses similar to severe cases of human COVID-19. The analysis of the structure and function of zebrafish and human Angiotensin-converting enzyme 2, the main human receptor for virus infection, presented remarkable sequence similarities. Moreover, bioinformatic analysis predicted protein-protein interaction of the Spike SARS-CoV-2 fragment and the Toll-like receptor pathway. It might help in the choice of future therapeutic pharmaceutical drugs to be studied. Based on the in vivo and in silico results presented here, we propose the zebrafish as a model for translational research into the safety of the vaccine and the immune response of the vertebrate organism to the SARS-CoV-2 virus.
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SciScore for 10.1101/2020.10.20.346262: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: The procedures were approved by the Ethics Committee (CEUA) of the XXXXXXXXXXXXXXXXXXXXXXXXX and registered under protocol number XXXXXXXX Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable rSpike was injected into two immunization sections in 20 zebrafish females (previously anesthetized with tricaine methanesulfonate (Sigma) - at a dose of 150 mg/L) at an interval of 7 days, with the aim of producing plasma antibodies. Table 2: Resources
Antibodies Sentences Resources After that, the primary antibodies were diluted in the aforementioned solution as follows: anti-Ly6G (1:300, Invitrogen, Clone RB6-8C5, Cat … SciScore for 10.1101/2020.10.20.346262: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: The procedures were approved by the Ethics Committee (CEUA) of the XXXXXXXXXXXXXXXXXXXXXXXXX and registered under protocol number XXXXXXXX Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable rSpike was injected into two immunization sections in 20 zebrafish females (previously anesthetized with tricaine methanesulfonate (Sigma) - at a dose of 150 mg/L) at an interval of 7 days, with the aim of producing plasma antibodies. Table 2: Resources
Antibodies Sentences Resources After that, the primary antibodies were diluted in the aforementioned solution as follows: anti-Ly6G (1:300, Invitrogen, Clone RB6-8C5, Cat 14-5931-81, host: rabbit), anti-AIF-1/Iba1 (1:300 anti-AIF-1/Iba1suggested: NoneSecondary antibodies for anti-Ly6G and anti-Iba1 primary antibodies were diluted as described above, as follows: anti-rabbit/Alexa488 (1:600, Invitrogen, Cat A21206, host: donkey) and anti-goat/Alexa594 (1:600, Invitrogen, Cat A11058, host: donkey). anti-Ly6Gsuggested: Noneanti-Iba1suggested: Noneanti-rabbit/Alexa488suggested: Noneanti-goat/Alexa594suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Zebrafish maintenance: Wild-type zebrafish from the AB line, and specific pathogen-free (SPF), were raised in Tecniplast Zebtec (Buguggiate, Italy) and maintained in the zebrafish housing systems in the XXXXXXXXXXXXXXX facilities. ABsuggested: RRID:BDSC_203)Software and Algorithms Sentences Resources Zebrafish were kept in 3.5 L polycarbonate tanks and fed three times a day with Gemma micro by Skretting (Stavanger, Norway). Gemmasuggested: (Gemma, RRID:SCR_008007)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 41. 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.
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