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<sup>+</sup> 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 fusio...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Vaccines |
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| Online Access: | https://www.mdpi.com/2076-393X/10/7/1060 |
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| author | Francesco Manfredi Chiara Chiozzini Flavia Ferrantelli Patrizia Leone Andrea Giovannelli Massimo Sanchez Maurizio Federico |
| author_facet | Francesco Manfredi Chiara Chiozzini Flavia Ferrantelli Patrizia Leone Andrea Giovannelli Massimo Sanchez Maurizio Federico |
| author_sort | Francesco Manfredi |
| collection | DOAJ |
| description | We propose an innovative anti-SARS-CoV-2 immune strategy based on extracellular vesicles (EVs) inducing an anti-SARS-CoV-2 N CD8<sup>+</sup> 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<sup>mut</sup>, 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<sup>mut</sup> or a deletion mutant of Nef<sup>mut</sup> referred to as Nef<sup>mut</sup>PL. The analysis of in vitro-produced EVs has supported the uploading of N protein when fused with truncated Nef<sup>mut</sup>. Mice injected with DNA vectors expressed each fusion protein developed robust SARS-CoV-2 N-specific CD8<sup>+</sup> 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. |
| first_indexed | 2024-03-09T10:10:11Z |
| format | Article |
| id | doaj.art-27a49e3cb82f43e88afcf14b2be30919 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-09T10:10:11Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-27a49e3cb82f43e88afcf14b2be309192023-12-01T22:46:15ZengMDPI AGVaccines2076-393X2022-06-01107106010.3390/vaccines10071060Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral ProteinFrancesco Manfredi0Chiara Chiozzini1Flavia Ferrantelli2Patrizia Leone3Andrea Giovannelli4Massimo Sanchez5Maurizio Federico6National Center for Global Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyNational Center for Global Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyNational Center for Global Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyNational Center for Global Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyNational Center for Animal Experimentation and Welfare, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyCore Facilities, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyNational Center for Global Health, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, ItalyWe propose an innovative anti-SARS-CoV-2 immune strategy based on extracellular vesicles (EVs) inducing an anti-SARS-CoV-2 N CD8<sup>+</sup> 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<sup>mut</sup>, 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<sup>mut</sup> or a deletion mutant of Nef<sup>mut</sup> referred to as Nef<sup>mut</sup>PL. The analysis of in vitro-produced EVs has supported the uploading of N protein when fused with truncated Nef<sup>mut</sup>. Mice injected with DNA vectors expressed each fusion protein developed robust SARS-CoV-2 N-specific CD8<sup>+</sup> 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.https://www.mdpi.com/2076-393X/10/7/1060extracellular vesiclesSARS-CoV-2CTL immunityHIV-1 NefCD107a |
| spellingShingle | Francesco Manfredi Chiara Chiozzini Flavia Ferrantelli Patrizia Leone Andrea Giovannelli Massimo Sanchez Maurizio Federico Activation of Anti-SARS-CoV-2 Human CTLs by Extracellular Vesicles Engineered with the N Viral Protein Vaccines extracellular vesicles SARS-CoV-2 CTL immunity HIV-1 Nef CD107a |
| 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 | extracellular vesicles SARS-CoV-2 CTL immunity HIV-1 Nef CD107a |
| url | https://www.mdpi.com/2076-393X/10/7/1060 |
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