A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines

IntroductionAs paradigmatically shown by SARS-CoV-2 vaccines, nucleic acids-based vaccines represent powerful tools to rapidly tackle fast emerging pathogens limiting their spread in human populations and/or reducing the health impact in affected patients. Compared with RNA vaccines, DNA vaccines of...

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Main Authors: Olivia Costantina Demurtas, Flavia Novelli, Doriana Triggiani, Caterina Merla, Emanuela Pasquali, Silvia Massa, Rosella Franconi, Claudio Pioli
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Medical Technology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmedt.2025.1597179/full
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author Olivia Costantina Demurtas
Flavia Novelli
Doriana Triggiani
Caterina Merla
Emanuela Pasquali
Silvia Massa
Rosella Franconi
Claudio Pioli
author_facet Olivia Costantina Demurtas
Flavia Novelli
Doriana Triggiani
Caterina Merla
Emanuela Pasquali
Silvia Massa
Rosella Franconi
Claudio Pioli
author_sort Olivia Costantina Demurtas
collection DOAJ
description IntroductionAs paradigmatically shown by SARS-CoV-2 vaccines, nucleic acids-based vaccines represent powerful tools to rapidly tackle fast emerging pathogens limiting their spread in human populations and/or reducing the health impact in affected patients. Compared with RNA vaccines, DNA vaccines offer higher stability and amenability to fast development due to tailor-made design of several candidates at a time for (pre)clinical settings. However, their scarce immunogenicity represents an important drawback, requiring technological strategies to enhance cellular uptake, protein expression and increase the ability to induce an immune response.MethodsWe investigated the effects of combining a plant secretory signal sequence of the PolyGalacturonase-Inhibiting Protein (PGIP) from Phaseolus vulgaris with electro-gene transfer (EGT), a technology that increases DNA delivery, on the immune response induced by different SARS-CoV2 experimental DNA vaccines based on domains and peptides of the spike (S), membrane (M) and nucleocapsid (N) proteins.Results and discussionAll the DNA constructs resulted in protein expression in vitro and in the induction of both antibody and CD4 and CD8T cell responses in mice. EGT significantly increased DNA constructs immunogenicity, especially for the induction of antibody response, confirming its potential in DNA vaccination. Remarkably, constructs including the plant secretory signal sequence resulted to be highly expressed and triggered higher antibody and CD4T cell responses, highlighting that the combination of this sequence and EGT can be used to boost the immunogenicity of DNA-vaccine coded proteins, ultimately helping in their design.
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spelling doaj-art-89bfde1d691d4f25819720c447e31ea12025-08-20T03:49:59ZengFrontiers Media S.A.Frontiers in Medical Technology2673-31292025-07-01710.3389/fmedt.2025.15971791597179A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccinesOlivia Costantina Demurtas0Flavia Novelli1Doriana Triggiani2Caterina Merla3Emanuela Pasquali4Silvia Massa5Rosella Franconi6Claudio Pioli7Energy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Sustainable Agrofood Systems (SSPT-AGROS), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyEnergy and Sustainable Economic Development, Division of Biotechnologies (SSPT-BIOTEC), ENEA, Italian National Agency for New Technologies, Rome, ItalyIntroductionAs paradigmatically shown by SARS-CoV-2 vaccines, nucleic acids-based vaccines represent powerful tools to rapidly tackle fast emerging pathogens limiting their spread in human populations and/or reducing the health impact in affected patients. Compared with RNA vaccines, DNA vaccines offer higher stability and amenability to fast development due to tailor-made design of several candidates at a time for (pre)clinical settings. However, their scarce immunogenicity represents an important drawback, requiring technological strategies to enhance cellular uptake, protein expression and increase the ability to induce an immune response.MethodsWe investigated the effects of combining a plant secretory signal sequence of the PolyGalacturonase-Inhibiting Protein (PGIP) from Phaseolus vulgaris with electro-gene transfer (EGT), a technology that increases DNA delivery, on the immune response induced by different SARS-CoV2 experimental DNA vaccines based on domains and peptides of the spike (S), membrane (M) and nucleocapsid (N) proteins.Results and discussionAll the DNA constructs resulted in protein expression in vitro and in the induction of both antibody and CD4 and CD8T cell responses in mice. EGT significantly increased DNA constructs immunogenicity, especially for the induction of antibody response, confirming its potential in DNA vaccination. Remarkably, constructs including the plant secretory signal sequence resulted to be highly expressed and triggered higher antibody and CD4T cell responses, highlighting that the combination of this sequence and EGT can be used to boost the immunogenicity of DNA-vaccine coded proteins, ultimately helping in their design.https://www.frontiersin.org/articles/10.3389/fmedt.2025.1597179/fullDNA vaccinesplant secretory sequenceelectro-gene transfer (EGT)COVID-19SARS-CoV-2receptor-binding domain (RBD)
spellingShingle Olivia Costantina Demurtas
Flavia Novelli
Doriana Triggiani
Caterina Merla
Emanuela Pasquali
Silvia Massa
Rosella Franconi
Claudio Pioli
A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines
Frontiers in Medical Technology
DNA vaccines
plant secretory sequence
electro-gene transfer (EGT)
COVID-19
SARS-CoV-2
receptor-binding domain (RBD)
title A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines
title_full A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines
title_fullStr A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines
title_full_unstemmed A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines
title_short A plant secretory sequence enhances immunogenicity of electroporated COVID-19 DNA vaccines
title_sort plant secretory sequence enhances immunogenicity of electroporated covid 19 dna vaccines
topic DNA vaccines
plant secretory sequence
electro-gene transfer (EGT)
COVID-19
SARS-CoV-2
receptor-binding domain (RBD)
url https://www.frontiersin.org/articles/10.3389/fmedt.2025.1597179/full
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