Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach

Background: The rapid advancement of the pandemic caused by SARS-CoV-2 and its variants reinforced the importance of developing easy-to-edit vaccines with fast production, such as multi-epitope DNA vaccines. The present study aimed to construct a synthetic antigen multi-epitope SARS-CoV-2 to produce...

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Main Authors: Maria da Conceição Viana Invenção, Larissa Silva de Macêdo, Ingrid Andrêssa de Moura, Lucas Alexandre Barbosa de Oliveira Santos, Benigno Cristofer Flores Espinoza, Samara Sousa de Pinho, Lígia Rosa Sales Leal, Daffany Luana dos Santos, Bianca de França São Marcos, Carolina Elsztein, Georon Ferreira de Sousa, Guilherme Antonio de Souza-Silva, Bárbara Rafaela da Silva Barros, Leonardo Carvalho de Oliveira Cruz, Julliano Matheus de Lima Maux, Jacinto da Costa Silva Neto, Cristiane Moutinho Lagos de Melo, Anna Jéssica Duarte Silva, Marcus Vinicius de Aragão Batista, Antonio Carlos de Freitas
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Language:English
Published: MDPI AG 2025-01-01
Series:Vaccines
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Online Access:https://www.mdpi.com/2076-393X/13/2/149
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author Maria da Conceição Viana Invenção
Larissa Silva de Macêdo
Ingrid Andrêssa de Moura
Lucas Alexandre Barbosa de Oliveira Santos
Benigno Cristofer Flores Espinoza
Samara Sousa de Pinho
Lígia Rosa Sales Leal
Daffany Luana dos Santos
Bianca de França São Marcos
Carolina Elsztein
Georon Ferreira de Sousa
Guilherme Antonio de Souza-Silva
Bárbara Rafaela da Silva Barros
Leonardo Carvalho de Oliveira Cruz
Julliano Matheus de Lima Maux
Jacinto da Costa Silva Neto
Cristiane Moutinho Lagos de Melo
Anna Jéssica Duarte Silva
Marcus Vinicius de Aragão Batista
Antonio Carlos de Freitas
author_facet Maria da Conceição Viana Invenção
Larissa Silva de Macêdo
Ingrid Andrêssa de Moura
Lucas Alexandre Barbosa de Oliveira Santos
Benigno Cristofer Flores Espinoza
Samara Sousa de Pinho
Lígia Rosa Sales Leal
Daffany Luana dos Santos
Bianca de França São Marcos
Carolina Elsztein
Georon Ferreira de Sousa
Guilherme Antonio de Souza-Silva
Bárbara Rafaela da Silva Barros
Leonardo Carvalho de Oliveira Cruz
Julliano Matheus de Lima Maux
Jacinto da Costa Silva Neto
Cristiane Moutinho Lagos de Melo
Anna Jéssica Duarte Silva
Marcus Vinicius de Aragão Batista
Antonio Carlos de Freitas
author_sort Maria da Conceição Viana Invenção
collection DOAJ
description Background: The rapid advancement of the pandemic caused by SARS-CoV-2 and its variants reinforced the importance of developing easy-to-edit vaccines with fast production, such as multi-epitope DNA vaccines. The present study aimed to construct a synthetic antigen multi-epitope SARS-CoV-2 to produce a DNA vaccine. Methods: A database of previously predicted Spike and Nucleocapsid protein epitopes was created, and these epitopes were analyzed for immunogenicity, conservation, population coverage, and molecular docking. Results: A synthetic antigen with 15 epitopes considered immunogenic, conserved even in the face of variants and that were able to anchor themselves in the appropriate HLA site, together had more than 90% worldwide coverage. A multi-epitope construct was developed with the sequences of these peptides separated from each other by linkers, cloned into the pVAX1 vector. This construct was evaluated in vivo as a DNA vaccine and elicited T CD4+ and T CD8+ cell expansion in the blood and spleen. In hematological analyses, there was an increase in lymphocytes, monocytes, and neutrophils between the two doses. Furthermore, based on histopathological analysis, the vaccines did not cause any damage to the organs analyzed. Conclusions: The present study generated a multi-epitope synthetic vaccine antigen capable of generating antibody-mediated and cellular immune responses.
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spelling doaj-art-528cce0b048e4aee96b5653dc5d82c5d2025-08-20T03:12:16ZengMDPI AGVaccines2076-393X2025-01-0113214910.3390/vaccines13020149Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo ApproachMaria da Conceição Viana Invenção0Larissa Silva de Macêdo1Ingrid Andrêssa de Moura2Lucas Alexandre Barbosa de Oliveira Santos3Benigno Cristofer Flores Espinoza4Samara Sousa de Pinho5Lígia Rosa Sales Leal6Daffany Luana dos Santos7Bianca de França São Marcos8Carolina Elsztein9Georon Ferreira de Sousa10Guilherme Antonio de Souza-Silva11Bárbara Rafaela da Silva Barros12Leonardo Carvalho de Oliveira Cruz13Julliano Matheus de Lima Maux14Jacinto da Costa Silva Neto15Cristiane Moutinho Lagos de Melo16Anna Jéssica Duarte Silva17Marcus Vinicius de Aragão Batista18Antonio Carlos de Freitas19Laboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Genetics and Biotechnology (GMBio), Department of Biology, Center for Biological and Health Sciences, Federal University of Sergipe, São Cristóvão 49100-000, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Immunological and Antitumor Analysis, Keizo Asami Immunopathology Laboratory, Department of Antibiotics, Bioscience Center, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Immunological and Antitumor Analysis, Keizo Asami Immunopathology Laboratory, Department of Antibiotics, Bioscience Center, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Immunological and Antitumor Analysis, Keizo Asami Immunopathology Laboratory, Department of Antibiotics, Bioscience Center, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Immunological and Antitumor Analysis, Keizo Asami Immunopathology Laboratory, Department of Antibiotics, Bioscience Center, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Cytological and Molecular Research, Department of Histology and Embriology, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Cytological and Molecular Research, Department of Histology and Embriology, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Immunological and Antitumor Analysis, Keizo Asami Immunopathology Laboratory, Department of Antibiotics, Bioscience Center, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilLaboratory of Molecular Genetics and Biotechnology (GMBio), Department of Biology, Center for Biological and Health Sciences, Federal University of Sergipe, São Cristóvão 49100-000, BrazilLaboratory of Molecular Studies and Experimental Therapy—LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, BrazilBackground: The rapid advancement of the pandemic caused by SARS-CoV-2 and its variants reinforced the importance of developing easy-to-edit vaccines with fast production, such as multi-epitope DNA vaccines. The present study aimed to construct a synthetic antigen multi-epitope SARS-CoV-2 to produce a DNA vaccine. Methods: A database of previously predicted Spike and Nucleocapsid protein epitopes was created, and these epitopes were analyzed for immunogenicity, conservation, population coverage, and molecular docking. Results: A synthetic antigen with 15 epitopes considered immunogenic, conserved even in the face of variants and that were able to anchor themselves in the appropriate HLA site, together had more than 90% worldwide coverage. A multi-epitope construct was developed with the sequences of these peptides separated from each other by linkers, cloned into the pVAX1 vector. This construct was evaluated in vivo as a DNA vaccine and elicited T CD4+ and T CD8+ cell expansion in the blood and spleen. In hematological analyses, there was an increase in lymphocytes, monocytes, and neutrophils between the two doses. Furthermore, based on histopathological analysis, the vaccines did not cause any damage to the organs analyzed. Conclusions: The present study generated a multi-epitope synthetic vaccine antigen capable of generating antibody-mediated and cellular immune responses.https://www.mdpi.com/2076-393X/13/2/149vaccinologyimmunoinformaticsvariants
spellingShingle Maria da Conceição Viana Invenção
Larissa Silva de Macêdo
Ingrid Andrêssa de Moura
Lucas Alexandre Barbosa de Oliveira Santos
Benigno Cristofer Flores Espinoza
Samara Sousa de Pinho
Lígia Rosa Sales Leal
Daffany Luana dos Santos
Bianca de França São Marcos
Carolina Elsztein
Georon Ferreira de Sousa
Guilherme Antonio de Souza-Silva
Bárbara Rafaela da Silva Barros
Leonardo Carvalho de Oliveira Cruz
Julliano Matheus de Lima Maux
Jacinto da Costa Silva Neto
Cristiane Moutinho Lagos de Melo
Anna Jéssica Duarte Silva
Marcus Vinicius de Aragão Batista
Antonio Carlos de Freitas
Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach
Vaccines
vaccinology
immunoinformatics
variants
title Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach
title_full Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach
title_fullStr Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach
title_full_unstemmed Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach
title_short Design and Immune Profile of Multi-Epitope Synthetic Antigen Vaccine Against SARS-CoV-2: An In Silico and In Vivo Approach
title_sort design and immune profile of multi epitope synthetic antigen vaccine against sars cov 2 an in silico and in vivo approach
topic vaccinology
immunoinformatics
variants
url https://www.mdpi.com/2076-393X/13/2/149
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