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Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein
We propose an innovative anti-SARS-CoV-2 immune strategy based on extracellular vesicles (EVs) inducing an anti-SARS-CoV-2 N CD8(+) T cytotoxic lymphocyte (CTL) immune response. We previously reported that the SARS-CoV-2 N protein can be uploaded at high levels in EVs upon fusion with Nef(mut), i.e....
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9318727/ https://www.ncbi.nlm.nih.gov/pubmed/35891224 http://dx.doi.org/10.3390/vaccines10071060 |
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author | Manfredi, Francesco Chiozzini, Chiara Ferrantelli, Flavia Leone, Patrizia Giovannelli, Andrea Sanchez, Massimo Federico, Maurizio |
author_facet | Manfredi, Francesco Chiozzini, Chiara Ferrantelli, Flavia Leone, Patrizia Giovannelli, Andrea Sanchez, Massimo Federico, Maurizio |
author_sort | Manfredi, Francesco |
collection | PubMed |
description | We propose an innovative anti-SARS-CoV-2 immune strategy based on extracellular vesicles (EVs) inducing an anti-SARS-CoV-2 N CD8(+) T cytotoxic lymphocyte (CTL) immune response. We previously reported that the SARS-CoV-2 N protein can be uploaded at high levels in EVs upon fusion with Nef(mut), i.e., a biologically inactive HIV-1 Nef mutant incorporating into EVs at quite high levels. Here, we analyze the immunogenic properties in human cells of EVs engineered with SARS-CoV-2 N fused at the C-terminus of either Nef(mut) or a deletion mutant of Nef(mut) referred to as Nef(mut)PL. The analysis of in vitro-produced EVs has supported the uploading of N protein when fused with truncated Nef(mut). Mice injected with DNA vectors expressed each fusion protein developed robust SARS-CoV-2 N-specific CD8(+) T cell immune responses. When ex vivo human dendritic cells were challenged with EVs engineered with either fusion products, the induction of a robust N-specific CTL activity, as evaluated by both CD107a and trogocytosis assays, was observed. Through these data we achieved the proof-of-principle that engineered EVs can be instrumental to elicit anti-SARS-CoV-2 CTL immune response in human cells. This achievement represents a mandatory step towards the upcoming experimentations in pre-clinical models focused on intranasal administration of N-engineered EVs. |
format | Online Article Text |
id | pubmed-9318727 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-93187272022-07-27 Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein Manfredi, Francesco Chiozzini, Chiara Ferrantelli, Flavia Leone, Patrizia Giovannelli, Andrea Sanchez, Massimo Federico, Maurizio Vaccines (Basel) Article We propose an innovative anti-SARS-CoV-2 immune strategy based on extracellular vesicles (EVs) inducing an anti-SARS-CoV-2 N CD8(+) T cytotoxic lymphocyte (CTL) immune response. We previously reported that the SARS-CoV-2 N protein can be uploaded at high levels in EVs upon fusion with Nef(mut), i.e., a biologically inactive HIV-1 Nef mutant incorporating into EVs at quite high levels. Here, we analyze the immunogenic properties in human cells of EVs engineered with SARS-CoV-2 N fused at the C-terminus of either Nef(mut) or a deletion mutant of Nef(mut) referred to as Nef(mut)PL. The analysis of in vitro-produced EVs has supported the uploading of N protein when fused with truncated Nef(mut). Mice injected with DNA vectors expressed each fusion protein developed robust SARS-CoV-2 N-specific CD8(+) T cell immune responses. When ex vivo human dendritic cells were challenged with EVs engineered with either fusion products, the induction of a robust N-specific CTL activity, as evaluated by both CD107a and trogocytosis assays, was observed. Through these data we achieved the proof-of-principle that engineered EVs can be instrumental to elicit anti-SARS-CoV-2 CTL immune response in human cells. This achievement represents a mandatory step towards the upcoming experimentations in pre-clinical models focused on intranasal administration of N-engineered EVs. MDPI 2022-06-30 /pmc/articles/PMC9318727/ /pubmed/35891224 http://dx.doi.org/10.3390/vaccines10071060 Text en © 2022 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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Manfredi, Francesco Chiozzini, Chiara Ferrantelli, Flavia Leone, Patrizia Giovannelli, Andrea Sanchez, Massimo Federico, Maurizio Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein |
title | Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein |
title_full | Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein |
title_fullStr | Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein |
title_full_unstemmed | Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein |
title_short | Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein |
title_sort | activation of anti-sars-cov-2 human ctls by extracellular vesicles engineered with the n viral protein |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9318727/ https://www.ncbi.nlm.nih.gov/pubmed/35891224 http://dx.doi.org/10.3390/vaccines10071060 |
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