The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles
Intramuscular injection of DNA vectors expressing the extracellular vesicle (EV)-anchoring protein Nef<sup>mut</sup> fused at its C-terminus to viral and tumor antigens elicit a potent, effective, and anti-tolerogenic CD8<sup>+</sup> T cell immunity against the heterologous a...
| Glavni autori: | , , , , , , |
|---|---|
| Format: | Članak |
| Jezik: | English |
| Izdano: |
MDPI AG
2021-04-01
|
| Serija: | Vaccines |
| Teme: | |
| Online pristup: | https://www.mdpi.com/2076-393X/9/4/373 |
| _version_ | 1827695353248350208 |
|---|---|
| author | Chiara Chiozzini Francesco Manfredi Flavia Ferrantelli Patrizia Leone Andrea Giovannelli Eleonora Olivetta Maurizio Federico |
| author_facet | Chiara Chiozzini Francesco Manfredi Flavia Ferrantelli Patrizia Leone Andrea Giovannelli Eleonora Olivetta Maurizio Federico |
| author_sort | Chiara Chiozzini |
| collection | DOAJ |
| description | Intramuscular injection of DNA vectors expressing the extracellular vesicle (EV)-anchoring protein Nef<sup>mut</sup> fused at its C-terminus to viral and tumor antigens elicit a potent, effective, and anti-tolerogenic CD8<sup>+</sup> T cell immunity against the heterologous antigen. The immune response is induced through the production of EVs incorporating Nef<sup>mut</sup>-derivatives released by muscle cells. In the perspective of a possible translation into the clinic of the Nef<sup>mut</sup>-based vaccine platform, we aimed at increasing its safety profile by identifying the minimal part of Nef<sup>mut</sup> retaining the EV-anchoring protein property. We found that a C-terminal deletion of 29-amino acids did not affect the ability of Nef<sup>mut</sup> to associate with EVs. The EV-anchoring function was also preserved when antigens from both HPV16 (i.e., E6 and E7) and SARS-CoV-2 (i.e., S1 and S2) were fused to its C-terminus. Most important, the Nef<sup>mut</sup> C-terminal deletion did not affect levels, quality, and diffusion at distal sites of the antigen-specific CD8<sup>+</sup> T immunity. We concluded that the C-terminal Nef<sup>mut</sup> truncation does not influence stability, EV-anchoring, and CD8<sup>+</sup> T cell immunogenicity of the fused antigen. Hence, the C-terminal deleted Nef<sup>mut</sup> may represent a safer alternative to the full-length isoform for vaccines in humans. |
| first_indexed | 2024-03-10T12:25:13Z |
| format | Article |
| id | doaj.art-863f59beec024b64840311d45176b793 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-03-10T12:25:13Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-863f59beec024b64840311d45176b7932023-11-21T15:07:51ZengMDPI AGVaccines2076-393X2021-04-019437310.3390/vaccines9040373The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular VesiclesChiara Chiozzini0Francesco Manfredi1Flavia Ferrantelli2Patrizia Leone3Andrea Giovannelli4Eleonora Olivetta5Maurizio 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, 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, ItalyIntramuscular injection of DNA vectors expressing the extracellular vesicle (EV)-anchoring protein Nef<sup>mut</sup> fused at its C-terminus to viral and tumor antigens elicit a potent, effective, and anti-tolerogenic CD8<sup>+</sup> T cell immunity against the heterologous antigen. The immune response is induced through the production of EVs incorporating Nef<sup>mut</sup>-derivatives released by muscle cells. In the perspective of a possible translation into the clinic of the Nef<sup>mut</sup>-based vaccine platform, we aimed at increasing its safety profile by identifying the minimal part of Nef<sup>mut</sup> retaining the EV-anchoring protein property. We found that a C-terminal deletion of 29-amino acids did not affect the ability of Nef<sup>mut</sup> to associate with EVs. The EV-anchoring function was also preserved when antigens from both HPV16 (i.e., E6 and E7) and SARS-CoV-2 (i.e., S1 and S2) were fused to its C-terminus. Most important, the Nef<sup>mut</sup> C-terminal deletion did not affect levels, quality, and diffusion at distal sites of the antigen-specific CD8<sup>+</sup> T immunity. We concluded that the C-terminal Nef<sup>mut</sup> truncation does not influence stability, EV-anchoring, and CD8<sup>+</sup> T cell immunogenicity of the fused antigen. Hence, the C-terminal deleted Nef<sup>mut</sup> may represent a safer alternative to the full-length isoform for vaccines in humans.https://www.mdpi.com/2076-393X/9/4/373extracellular vesiclesHIV-1 NefDNA immunizationCD8<sup>+</sup> T cell immunityHPV vaccinesSARS-CoV-2 vaccines |
| spellingShingle | Chiara Chiozzini Francesco Manfredi Flavia Ferrantelli Patrizia Leone Andrea Giovannelli Eleonora Olivetta Maurizio Federico The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles Vaccines extracellular vesicles HIV-1 Nef DNA immunization CD8<sup>+</sup> T cell immunity HPV vaccines SARS-CoV-2 vaccines |
| title | The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles |
| title_full | The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles |
| title_fullStr | The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles |
| title_full_unstemmed | The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles |
| title_short | The C-Terminal Domain of Nef<sup>mut</sup> Is Dispensable for the CD8<sup>+</sup> T Cell Immunogenicity of In Vivo Engineered Extracellular Vesicles |
| title_sort | c terminal domain of nef sup mut sup is dispensable for the cd8 sup sup t cell immunogenicity of in vivo engineered extracellular vesicles |
| topic | extracellular vesicles HIV-1 Nef DNA immunization CD8<sup>+</sup> T cell immunity HPV vaccines SARS-CoV-2 vaccines |
| url | https://www.mdpi.com/2076-393X/9/4/373 |
| work_keys_str_mv | AT chiarachiozzini thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT francescomanfredi thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT flaviaferrantelli thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT patrizialeone thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT andreagiovannelli thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT eleonoraolivetta thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT mauriziofederico thecterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT chiarachiozzini cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT francescomanfredi cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT flaviaferrantelli cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT patrizialeone cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT andreagiovannelli cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT eleonoraolivetta cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles AT mauriziofederico cterminaldomainofnefsupmutsupisdispensableforthecd8supsuptcellimmunogenicityofinvivoengineeredextracellularvesicles |