Bacterial outer membrane vesicles (OMVs) represent an interesting vaccine platform for their built-in adjuvanticity and simplicity of production process. Moreover, OMVs can be decorated with foreign antigens using different synthetic biology approaches. However, the optimal OMV engineering strategy, which should guarantee the OMV compartmentalization of most heterologous antigens in quantities high enough to elicit protective immune responses, remains to be validated. In this work we exploited the lipoprotein transport pathway to engineer OMVs with foreign proteins. Using 5 Staphylococcus aureus protective antigens expressed in Escherichia coli as fusions to a lipoprotein leader sequence, we demonstrated that all 5 antigens accumulated in the vesicular compartment at a concentration ranging from 5 to 20% of total OMV proteins, suggesting that antigen lipidation could be a universal approach for OMV manipulation. Engineered OMVs elicited high, saturating antigen-specific antibody titers when administered to mice in quantities as low as 0.2 μg/dose. Moreover, the expression of lipidated antigens in E. coli BL21(DE3)ΔompAΔmsbBΔpagP was shown to affect the lipopolysaccharide structure, with the result that the TLR4 agonist activity of OMVs was markedly reduced. These results, together with the potent protective activity of engineered OMVs observed in mice challenged with S. aureus Newman strain, makes the 5-combo-OMVs a promising vaccine candidate to be tested in clinics.

Bacterial outer membrane vesicles engineered with lipidated antigens as a platform for Staphylococcus aureus vaccine / Irene, Carmela; Fantappiè, Laura; Caproni, Elena; Zerbini, Francesca; Anesi, Andrea; Tomasi, Michele; Zanella, Ilaria; Stupia, Simone; Prete, Stefano; Valensin, Silvia; Koenig, Enrico; Frattini, Luca; Gagliardi, Assunta; Isaac, Samine J.; Grandi, Alberto; Guella, Graziano; Grandi, Guido. - In: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. - ISSN 0027-8424. - STAMPA. - 116:43(2019), pp. 21780-21788. [10.1073/pnas.1905112116]

Bacterial outer membrane vesicles engineered with lipidated antigens as a platform for Staphylococcus aureus vaccine

Irene, Carmela;Fantappiè, Laura;Caproni, Elena;Zerbini, Francesca;Anesi, Andrea;Tomasi, Michele;Zanella, Ilaria;Koenig, Enrico;Frattini, Luca;Gagliardi, Assunta;Isaac, Samine J.;Grandi, Alberto;Guella, Graziano;Grandi, Guido
2019-01-01

Abstract

Bacterial outer membrane vesicles (OMVs) represent an interesting vaccine platform for their built-in adjuvanticity and simplicity of production process. Moreover, OMVs can be decorated with foreign antigens using different synthetic biology approaches. However, the optimal OMV engineering strategy, which should guarantee the OMV compartmentalization of most heterologous antigens in quantities high enough to elicit protective immune responses, remains to be validated. In this work we exploited the lipoprotein transport pathway to engineer OMVs with foreign proteins. Using 5 Staphylococcus aureus protective antigens expressed in Escherichia coli as fusions to a lipoprotein leader sequence, we demonstrated that all 5 antigens accumulated in the vesicular compartment at a concentration ranging from 5 to 20% of total OMV proteins, suggesting that antigen lipidation could be a universal approach for OMV manipulation. Engineered OMVs elicited high, saturating antigen-specific antibody titers when administered to mice in quantities as low as 0.2 μg/dose. Moreover, the expression of lipidated antigens in E. coli BL21(DE3)ΔompAΔmsbBΔpagP was shown to affect the lipopolysaccharide structure, with the result that the TLR4 agonist activity of OMVs was markedly reduced. These results, together with the potent protective activity of engineered OMVs observed in mice challenged with S. aureus Newman strain, makes the 5-combo-OMVs a promising vaccine candidate to be tested in clinics.
2019
43
Irene, Carmela; Fantappiè, Laura; Caproni, Elena; Zerbini, Francesca; Anesi, Andrea; Tomasi, Michele; Zanella, Ilaria; Stupia, Simone; Prete, Stefano;...espandi
Bacterial outer membrane vesicles engineered with lipidated antigens as a platform for Staphylococcus aureus vaccine / Irene, Carmela; Fantappiè, Laura; Caproni, Elena; Zerbini, Francesca; Anesi, Andrea; Tomasi, Michele; Zanella, Ilaria; Stupia, Simone; Prete, Stefano; Valensin, Silvia; Koenig, Enrico; Frattini, Luca; Gagliardi, Assunta; Isaac, Samine J.; Grandi, Alberto; Guella, Graziano; Grandi, Guido. - In: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. - ISSN 0027-8424. - STAMPA. - 116:43(2019), pp. 21780-21788. [10.1073/pnas.1905112116]
File in questo prodotto:
File Dimensione Formato  
Irene_2019.pdf

accesso aperto

Tipologia: Versione editoriale (Publisher’s layout)
Licenza: Creative commons
Dimensione 1.16 MB
Formato Adobe PDF
1.16 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11572/277653
Citazioni
  • ???jsp.display-item.citation.pmc??? 48
  • Scopus 73
  • ???jsp.display-item.citation.isi??? 68
  • OpenAlex ND
social impact