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Zebrafish studies on the vaccine candidate to COVID-19, the Spike protein: Production of antibody and adverse reaction

Bianca H Ventura Fernandes, Natália Martins Feitosa, Ana Paula Barbosa, Camila Gasque Bomfim, Anali M. B. Garnique, Francisco I. F. Gomes, Rafael T. Nakajima, Marco A. A. Belo, Silas Fernandes Eto, Dayanne Carla Fernandes, Guilherme Malafaia, Wilson G. Manrique, Gabriel Conde, Roberta R. C. Rosales, Iris Todeschini, Ilo Rivero, Edgar Llontop, German G. Sgro, Gabriel Umaji Oka, Natalia F Bueno, Fausto K. Ferraris, Mariana T. Q. de Magalhaes, Renata J. Medeiros, Juliana M. M Gomes, Mara de Souza Junqueira, Katia Conceição, Letícia G. Pontes, Antonio Condino-Neto, Andrea C. Perez, Leonardo J. G. Barcellos, Jose Dias Correa junior, Erick G. Dorlass, Niels O. S Camara, Edison Luiz Durigon, Fernando Q. Cunha, View ORCID ProfileRafael H. Nóbrega, Glaucia M. Machado-Santelli, Chuck Farah, Flávio P Veras, Jorge Galindo-Villegas, Leticia Costa-Lotufo, Thiago M. Cunha, Roger Chammas, Cristiane R. Guzzo, Luciani R Carvalho, Ives Charlie-Silva
doi: https://doi.org/10.1101/2020.10.20.346262
Bianca H Ventura Fernandes
1Laboratório de Controle Genético e Sanitário, Diretoria Técnica de Apoio ao Ensino e Pesquisa, Faculdade de Medicina da Universidade de São Paulo
32Disciplina de endocrinologia do departamento de Clinica medica e Laboratório de Hormônios e Genética Molecular, LIM 42
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Natália Martins Feitosa
2Integrated Laboratory of Translational Bioscience (LIBT), Institute of Biodiversity and Sustainability (NUPEM), Federal University of Rio de Janeiro (UFRJ)-Macaé, RJ, Brazil
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Ana Paula Barbosa
3Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil
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Camila Gasque Bomfim
3Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil
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Anali M. B. Garnique
4Department of Cell Biology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil
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Francisco I. F. Gomes
5Center of Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
6Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
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Rafael T. Nakajima
7Reproductive and Molecular Biology Group, Department of Morphology, Institute of Biosciences, São Paulo State University, Botucatu, São Paulo, Brazil
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Marco A. A. Belo
8Department of Preventive Veterinary Medicine, São Paulo State University, Jaboticabal, São Paulo, Brazil
9Laboratory of Animal Pharmacology and Toxicology, Brasil University, Descalvado, São Paulo, Brazil
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Silas Fernandes Eto
10Postgraduate Program in Health Sciences - PROCISA, Federal University of Roraima, Brazil
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Dayanne Carla Fernandes
11Immunochemistry Laboratory, Butantan Institute, São Paulo, Brazil
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Guilherme Malafaia
12Biological Research Laboratory, Goiano Federal Institute – Urutaí Campus, GO, Brazil
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Wilson G. Manrique
13Aquaculture Health Research and Extension Group - GRUPESA, Aquaculture Health Laboratory - LABSA, Department of Veterinary Medicine, Federal University of Rondônia, Rolim de Moura campus, Rondônia, Brazil
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Gabriel Conde
8Department of Preventive Veterinary Medicine, São Paulo State University, Jaboticabal, São Paulo, Brazil
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Roberta R. C. Rosales
14Department of Cell and Molecular Biology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
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Iris Todeschini
15Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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Ilo Rivero
16Pontificia Universidade Catolica de Minas Gerais
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Edgar Llontop
15Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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German G. Sgro
15Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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Gabriel Umaji Oka
15Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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Natalia F Bueno
15Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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Fausto K. Ferraris
17Department of Pharmacology and Toxicology, Oswaldo Cruz Foundation - FIOCRUZ- Rio de Janeiro, Brazil
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Mariana T. Q. de Magalhaes
18Department of Biochemistry and Immunology, Institute of Biological Sciences – ICB, Federal University of Minas Gerais, Belo Horizonte, Brazil
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Renata J. Medeiros
19Laboratory of Physiology - Zebrafish INCQS/Fiocruz Facility, Department of Pharmacology and Toxicology – DFT, INCQS - National Institute for Quality Control in Health. Rio de Janeiro, Brazil
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Juliana M. M Gomes
20Transplantation Immunobiology Lab, Department of Immunology, Institute of Biomedical Sciences, Universidade de São Paulo, Brazil
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Mara de Souza Junqueira
21Center for Translational Research in Oncology, Cancer Institute of the State of São Paulo, Faculty of Medicine, University of São Paulo, São Paulo. Brazil
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Katia Conceição
22Laboratory of Peptide Biochemistry, Federal University of São Paulo, Brazil
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Letícia G. Pontes
23Laboratory of Human Immunology, Department Immunology, Institute Biomedical Sciences, University Sao Paulo, Sao Paulo – Brazil
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Antonio Condino-Neto
23Laboratory of Human Immunology, Department Immunology, Institute Biomedical Sciences, University Sao Paulo, Sao Paulo – Brazil
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Andrea C. Perez
24Department of Pharmacology, Universidade Federal de Minas Gerais, Brazil
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Leonardo J. G. Barcellos
25Graduate Program of Pharmacology, Federal University of Santa Maria, Brazil
26Laboratory of Fish Physiology, Graduate Program of Bioexperimentation and of Environmental Sciences. University of Passo Fundo, Brazil
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Jose Dias Correa junior
27Laboratorio do Estudo da Interacao Quimico Biologica e da Reproducao Animal, LIQBRA, Bloco O3,174, Departamento de Morfologia Instituto de Ciencias Biologicas, Universidade Federal de Minas Gerais
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Erick G. Dorlass
3Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil
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Niels O. S Camara
20Transplantation Immunobiology Lab, Department of Immunology, Institute of Biomedical Sciences, Universidade de São Paulo, Brazil
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Edison Luiz Durigon
3Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil
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Fernando Q. Cunha
5Center of Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
6Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
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Rafael H. Nóbrega
7Reproductive and Molecular Biology Group, Department of Morphology, Institute of Biosciences, São Paulo State University, Botucatu, São Paulo, Brazil
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  • ORCID record for Rafael H. Nóbrega
Glaucia M. Machado-Santelli
28Department of Cell Biology, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, Brazil
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Chuck Farah
15Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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Flávio P Veras
5Center of Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
6Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
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Jorge Galindo-Villegas
29Faculty of Biosciences and Aquaculture, Nord University, 8049 Bodø, Norway
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Leticia Costa-Lotufo
30Department of Pharmacology, Institute of Biomedical Sciences, Universidade de Sao Paulo, Brazil
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Thiago M. Cunha
5Center of Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
6Department of Pharmacology, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil
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Roger Chammas
1Laboratório de Controle Genético e Sanitário, Diretoria Técnica de Apoio ao Ensino e Pesquisa, Faculdade de Medicina da Universidade de São Paulo
31Centro de Investigação Translacional em Oncologia, Instituto do Câncer do Estado de São Paulo, Faculdade de Medicina da Universidade de São Paulo
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Cristiane R. Guzzo
3Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil
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Luciani R Carvalho
1Laboratório de Controle Genético e Sanitário, Diretoria Técnica de Apoio ao Ensino e Pesquisa, Faculdade de Medicina da Universidade de São Paulo
32Disciplina de endocrinologia do departamento de Clinica medica e Laboratório de Hormônios e Genética Molecular, LIM 42
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Ives Charlie-Silva
30Department of Pharmacology, Institute of Biomedical Sciences, Universidade de Sao Paulo, Brazil
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  • For correspondence: Charliesilva4@hotmail.com
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Summary

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.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Zebrafish studies on the vaccine candidate to COVID-19, the Spike protein: Production of antibody and adverse reaction
Bianca H Ventura Fernandes, Natália Martins Feitosa, Ana Paula Barbosa, Camila Gasque Bomfim, Anali M. B. Garnique, Francisco I. F. Gomes, Rafael T. Nakajima, Marco A. A. Belo, Silas Fernandes Eto, Dayanne Carla Fernandes, Guilherme Malafaia, Wilson G. Manrique, Gabriel Conde, Roberta R. C. Rosales, Iris Todeschini, Ilo Rivero, Edgar Llontop, German G. Sgro, Gabriel Umaji Oka, Natalia F Bueno, Fausto K. Ferraris, Mariana T. Q. de Magalhaes, Renata J. Medeiros, Juliana M. M Gomes, Mara de Souza Junqueira, Katia Conceição, Letícia G. Pontes, Antonio Condino-Neto, Andrea C. Perez, Leonardo J. G. Barcellos, Jose Dias Correa junior, Erick G. Dorlass, Niels O. S Camara, Edison Luiz Durigon, Fernando Q. Cunha, Rafael H. Nóbrega, Glaucia M. Machado-Santelli, Chuck Farah, Flávio P Veras, Jorge Galindo-Villegas, Leticia Costa-Lotufo, Thiago M. Cunha, Roger Chammas, Cristiane R. Guzzo, Luciani R Carvalho, Ives Charlie-Silva
bioRxiv 2020.10.20.346262; doi: https://doi.org/10.1101/2020.10.20.346262
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Zebrafish studies on the vaccine candidate to COVID-19, the Spike protein: Production of antibody and adverse reaction
Bianca H Ventura Fernandes, Natália Martins Feitosa, Ana Paula Barbosa, Camila Gasque Bomfim, Anali M. B. Garnique, Francisco I. F. Gomes, Rafael T. Nakajima, Marco A. A. Belo, Silas Fernandes Eto, Dayanne Carla Fernandes, Guilherme Malafaia, Wilson G. Manrique, Gabriel Conde, Roberta R. C. Rosales, Iris Todeschini, Ilo Rivero, Edgar Llontop, German G. Sgro, Gabriel Umaji Oka, Natalia F Bueno, Fausto K. Ferraris, Mariana T. Q. de Magalhaes, Renata J. Medeiros, Juliana M. M Gomes, Mara de Souza Junqueira, Katia Conceição, Letícia G. Pontes, Antonio Condino-Neto, Andrea C. Perez, Leonardo J. G. Barcellos, Jose Dias Correa junior, Erick G. Dorlass, Niels O. S Camara, Edison Luiz Durigon, Fernando Q. Cunha, Rafael H. Nóbrega, Glaucia M. Machado-Santelli, Chuck Farah, Flávio P Veras, Jorge Galindo-Villegas, Leticia Costa-Lotufo, Thiago M. Cunha, Roger Chammas, Cristiane R. Guzzo, Luciani R Carvalho, Ives Charlie-Silva
bioRxiv 2020.10.20.346262; doi: https://doi.org/10.1101/2020.10.20.346262

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