Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs

Most advanced vaccines against severe acute respiratory syndrome coronavirus (SARS-CoV)-2 are designed to induce antibodies against spike (S) protein. Differently, we developed an original strategy to induce CD8(+) T cytotoxic lymphocyte (CTL) immunity based on in vivo engineering of extracellular v...

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Autores principales: Ferrantelli, Flavia, Chiozzini, Chiara, Manfredi, Francesco, Giovannelli, Andrea, Leone, Patrizia, Federico, Maurizio
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999804/
https://www.ncbi.nlm.nih.gov/pubmed/33801926
http://dx.doi.org/10.3390/vaccines9030240
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author Ferrantelli, Flavia
Chiozzini, Chiara
Manfredi, Francesco
Giovannelli, Andrea
Leone, Patrizia
Federico, Maurizio
author_facet Ferrantelli, Flavia
Chiozzini, Chiara
Manfredi, Francesco
Giovannelli, Andrea
Leone, Patrizia
Federico, Maurizio
author_sort Ferrantelli, Flavia
collection PubMed
description Most advanced vaccines against severe acute respiratory syndrome coronavirus (SARS-CoV)-2 are designed to induce antibodies against spike (S) protein. Differently, we developed an original strategy to induce CD8(+) T cytotoxic lymphocyte (CTL) immunity based on in vivo engineering of extracellular vesicles (EVs). This is a new vaccination approach based on intramuscular injection of DNA expression vectors coding for a biologically inactive HIV-1 Nef protein (Nef(mut)) with an unusually high efficiency of incorporation into EVs, even when foreign polypeptides are fused to its C-terminus. Nanovesicles containing Nef(mut)-fused antigens released by muscle cells can freely circulate into the body and are internalized by antigen-presenting cells. Therefore, EV-associated antigens can be cross-presented to prime antigen-specific CD8(+) T-cells. To apply this technology to a strategy of anti-SARS-CoV-2 vaccine, we designed DNA vectors expressing the products of fusion between Nef(mut) and different viral antigens, namely N- and C-terminal moieties of S (referred to as S1 and S2), M, and N. We provided evidence that all fusion products are efficiently uploaded in EVs. When the respective DNA vectors were injected in mice, a strong antigen-specific CD8(+) T cell immunity became detectable in spleens and, most important, in lung airways. Co-injection of DNA vectors expressing the diverse SARS-CoV-2 antigens resulted in additive immune responses in both spleen and lungs. Hence, DNA vectors expressing Nef(mut)-based fusion proteins can be proposed for new anti-SARS-CoV-2 vaccine strategies.
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spelling pubmed-79998042021-03-28 Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs Ferrantelli, Flavia Chiozzini, Chiara Manfredi, Francesco Giovannelli, Andrea Leone, Patrizia Federico, Maurizio Vaccines (Basel) Article Most advanced vaccines against severe acute respiratory syndrome coronavirus (SARS-CoV)-2 are designed to induce antibodies against spike (S) protein. Differently, we developed an original strategy to induce CD8(+) T cytotoxic lymphocyte (CTL) immunity based on in vivo engineering of extracellular vesicles (EVs). This is a new vaccination approach based on intramuscular injection of DNA expression vectors coding for a biologically inactive HIV-1 Nef protein (Nef(mut)) with an unusually high efficiency of incorporation into EVs, even when foreign polypeptides are fused to its C-terminus. Nanovesicles containing Nef(mut)-fused antigens released by muscle cells can freely circulate into the body and are internalized by antigen-presenting cells. Therefore, EV-associated antigens can be cross-presented to prime antigen-specific CD8(+) T-cells. To apply this technology to a strategy of anti-SARS-CoV-2 vaccine, we designed DNA vectors expressing the products of fusion between Nef(mut) and different viral antigens, namely N- and C-terminal moieties of S (referred to as S1 and S2), M, and N. We provided evidence that all fusion products are efficiently uploaded in EVs. When the respective DNA vectors were injected in mice, a strong antigen-specific CD8(+) T cell immunity became detectable in spleens and, most important, in lung airways. Co-injection of DNA vectors expressing the diverse SARS-CoV-2 antigens resulted in additive immune responses in both spleen and lungs. Hence, DNA vectors expressing Nef(mut)-based fusion proteins can be proposed for new anti-SARS-CoV-2 vaccine strategies. MDPI 2021-03-10 /pmc/articles/PMC7999804/ /pubmed/33801926 http://dx.doi.org/10.3390/vaccines9030240 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Ferrantelli, Flavia
Chiozzini, Chiara
Manfredi, Francesco
Giovannelli, Andrea
Leone, Patrizia
Federico, Maurizio
Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs
title Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs
title_full Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs
title_fullStr Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs
title_full_unstemmed Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs
title_short Simultaneous CD8(+) T-Cell Immune Response against SARS-Cov-2 S, M, and N Induced by Endogenously Engineered Extracellular Vesicles in Both Spleen and Lungs
title_sort simultaneous cd8(+) t-cell immune response against sars-cov-2 s, m, and n induced by endogenously engineered extracellular vesicles in both spleen and lungs
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999804/
https://www.ncbi.nlm.nih.gov/pubmed/33801926
http://dx.doi.org/10.3390/vaccines9030240
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