Decoration of outer membrane vesicles with multiple antigens using an autotransporter approach.

M.H. Daleke, T. Felix, Z. Soprova, C.M. ten Hagen-Jongman ten, D. Vikström, L. Majlessi, J. Beskers, F. Follman, K. de Punder, N.N. van der Wel, T. Baumgarten, T.V. Pham, S.R. Piersma, C.R. Jimenez, P. van Ulsen, J.W. de Gier, C. Leclerc, W.S. Jong, S. Luirink

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Outer membrane vesicles (OMVs) are spherical nanoparticles that naturally shed from Gram-negative bacteria. They are rich in immunostimulatory proteins and lipopolysaccharide but do not replicate, which increases their safety profile and renders them attractive vaccine vectors. By packaging foreign polypeptides in OMVs, specific immune responses can be raised toward heterologous antigens in the context of an intrinsic adjuvant. Antigens exposed at the vesicle surface have been suggested to elicit protection superior to that from antigens concealed inside OMVs, but hitherto robust methods for targeting heterologous proteins to the OMV surface have been lacking. We have exploited our previously developed hemoglobin protease (Hbp) autotransporter platform for display of heterologous polypeptides at the OMV surface. One, two, or three of the Mycobacterium tuberculosis antigens ESAT6, Ag85B, and Rv2660c were targeted to the surface of Escherichia coli OMVs upon fusion to Hbp. Furthermore, a hypervesiculating ΔtolR ΔtolA derivative of attenuated Salmonella enterica serovar Typhimurium SL3261 was generated, enabling efficient release and purification of OMVs decorated with multiple heterologous antigens, exemplified by the M. tuberculosis antigens and epitopes from Chlamydia trachomatis major outer membrane protein (MOMP). Also, we showed that delivery of Salmonella OMVs displaying Ag85B to antigen-presenting cells in vitro results in processing and presentation of an epitope that is functionally recognized by Ag85B-specific T cell hybridomas. In conclusion, the Hbp platform mediates efficient display of (multiple) heterologous antigens, individually or combined within one molecule, at the surface of OMVs. Detection of antigen- specific immune responses upon vesicle-mediated delivery demonstrated the potential of our system for vaccine development. © 2014, American Society for Microbiology.
LanguageEnglish
Pages5854-5864
JournalApplied and Environmental Microbiology
Volume80
Issue number18
DOIs
Publication statusPublished - 2014

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vesicle
antigen
membrane
antigens
Antigens
Membranes
Heterophile Antigens
hemoglobin
proteinases
Mycobacterium tuberculosis
Hemoglobins
Peptide Hydrolases
epitopes
polypeptides
tuberculosis
immune response
Chlamydia trachomatis
vaccine
Epitopes
vector vaccines

Cite this

Daleke, M.H. ; Felix, T. ; Soprova, Z. ; ten Hagen-Jongman ten, C.M. ; Vikström, D. ; Majlessi, L. ; Beskers, J. ; Follman, F. ; de Punder, K. ; van der Wel, N.N. ; Baumgarten, T. ; Pham, T.V. ; Piersma, S.R. ; Jimenez, C.R. ; van Ulsen, P. ; de Gier, J.W. ; Leclerc, C. ; Jong, W.S. ; Luirink, S. / Decoration of outer membrane vesicles with multiple antigens using an autotransporter approach. In: Applied and Environmental Microbiology. 2014 ; Vol. 80, No. 18. pp. 5854-5864.
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title = "Decoration of outer membrane vesicles with multiple antigens using an autotransporter approach.",
abstract = "Outer membrane vesicles (OMVs) are spherical nanoparticles that naturally shed from Gram-negative bacteria. They are rich in immunostimulatory proteins and lipopolysaccharide but do not replicate, which increases their safety profile and renders them attractive vaccine vectors. By packaging foreign polypeptides in OMVs, specific immune responses can be raised toward heterologous antigens in the context of an intrinsic adjuvant. Antigens exposed at the vesicle surface have been suggested to elicit protection superior to that from antigens concealed inside OMVs, but hitherto robust methods for targeting heterologous proteins to the OMV surface have been lacking. We have exploited our previously developed hemoglobin protease (Hbp) autotransporter platform for display of heterologous polypeptides at the OMV surface. One, two, or three of the Mycobacterium tuberculosis antigens ESAT6, Ag85B, and Rv2660c were targeted to the surface of Escherichia coli OMVs upon fusion to Hbp. Furthermore, a hypervesiculating ΔtolR ΔtolA derivative of attenuated Salmonella enterica serovar Typhimurium SL3261 was generated, enabling efficient release and purification of OMVs decorated with multiple heterologous antigens, exemplified by the M. tuberculosis antigens and epitopes from Chlamydia trachomatis major outer membrane protein (MOMP). Also, we showed that delivery of Salmonella OMVs displaying Ag85B to antigen-presenting cells in vitro results in processing and presentation of an epitope that is functionally recognized by Ag85B-specific T cell hybridomas. In conclusion, the Hbp platform mediates efficient display of (multiple) heterologous antigens, individually or combined within one molecule, at the surface of OMVs. Detection of antigen- specific immune responses upon vesicle-mediated delivery demonstrated the potential of our system for vaccine development. {\circledC} 2014, American Society for Microbiology.",
author = "M.H. Daleke and T. Felix and Z. Soprova and {ten Hagen-Jongman ten}, C.M. and D. Vikstr{\"o}m and L. Majlessi and J. Beskers and F. Follman and {de Punder}, K. and {van der Wel}, N.N. and T. Baumgarten and T.V. Pham and S.R. Piersma and C.R. Jimenez and {van Ulsen}, P. and {de Gier}, J.W. and C. Leclerc and W.S. Jong and S. Luirink",
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Daleke, MH, Felix, T, Soprova, Z, ten Hagen-Jongman ten, CM, Vikström, D, Majlessi, L, Beskers, J, Follman, F, de Punder, K, van der Wel, NN, Baumgarten, T, Pham, TV, Piersma, SR, Jimenez, CR, van Ulsen, P, de Gier, JW, Leclerc, C, Jong, WS & Luirink, S 2014, 'Decoration of outer membrane vesicles with multiple antigens using an autotransporter approach.', Applied and Environmental Microbiology, vol. 80, no. 18, pp. 5854-5864. https://doi.org/10.1128/AEM.01941-14

Decoration of outer membrane vesicles with multiple antigens using an autotransporter approach. / Daleke, M.H.; Felix, T.; Soprova, Z.; ten Hagen-Jongman ten, C.M.; Vikström, D.; Majlessi, L.; Beskers, J.; Follman, F.; de Punder, K.; van der Wel, N.N.; Baumgarten, T.; Pham, T.V.; Piersma, S.R.; Jimenez, C.R.; van Ulsen, P.; de Gier, J.W.; Leclerc, C.; Jong, W.S.; Luirink, S.

In: Applied and Environmental Microbiology, Vol. 80, No. 18, 2014, p. 5854-5864.

Research output: Contribution to JournalArticleAcademicpeer-review

TY - JOUR

T1 - Decoration of outer membrane vesicles with multiple antigens using an autotransporter approach.

AU - Daleke, M.H.

AU - Felix, T.

AU - Soprova, Z.

AU - ten Hagen-Jongman ten, C.M.

AU - Vikström, D.

AU - Majlessi, L.

AU - Beskers, J.

AU - Follman, F.

AU - de Punder, K.

AU - van der Wel, N.N.

AU - Baumgarten, T.

AU - Pham, T.V.

AU - Piersma, S.R.

AU - Jimenez, C.R.

AU - van Ulsen, P.

AU - de Gier, J.W.

AU - Leclerc, C.

AU - Jong, W.S.

AU - Luirink, S.

PY - 2014

Y1 - 2014

N2 - Outer membrane vesicles (OMVs) are spherical nanoparticles that naturally shed from Gram-negative bacteria. They are rich in immunostimulatory proteins and lipopolysaccharide but do not replicate, which increases their safety profile and renders them attractive vaccine vectors. By packaging foreign polypeptides in OMVs, specific immune responses can be raised toward heterologous antigens in the context of an intrinsic adjuvant. Antigens exposed at the vesicle surface have been suggested to elicit protection superior to that from antigens concealed inside OMVs, but hitherto robust methods for targeting heterologous proteins to the OMV surface have been lacking. We have exploited our previously developed hemoglobin protease (Hbp) autotransporter platform for display of heterologous polypeptides at the OMV surface. One, two, or three of the Mycobacterium tuberculosis antigens ESAT6, Ag85B, and Rv2660c were targeted to the surface of Escherichia coli OMVs upon fusion to Hbp. Furthermore, a hypervesiculating ΔtolR ΔtolA derivative of attenuated Salmonella enterica serovar Typhimurium SL3261 was generated, enabling efficient release and purification of OMVs decorated with multiple heterologous antigens, exemplified by the M. tuberculosis antigens and epitopes from Chlamydia trachomatis major outer membrane protein (MOMP). Also, we showed that delivery of Salmonella OMVs displaying Ag85B to antigen-presenting cells in vitro results in processing and presentation of an epitope that is functionally recognized by Ag85B-specific T cell hybridomas. In conclusion, the Hbp platform mediates efficient display of (multiple) heterologous antigens, individually or combined within one molecule, at the surface of OMVs. Detection of antigen- specific immune responses upon vesicle-mediated delivery demonstrated the potential of our system for vaccine development. © 2014, American Society for Microbiology.

AB - Outer membrane vesicles (OMVs) are spherical nanoparticles that naturally shed from Gram-negative bacteria. They are rich in immunostimulatory proteins and lipopolysaccharide but do not replicate, which increases their safety profile and renders them attractive vaccine vectors. By packaging foreign polypeptides in OMVs, specific immune responses can be raised toward heterologous antigens in the context of an intrinsic adjuvant. Antigens exposed at the vesicle surface have been suggested to elicit protection superior to that from antigens concealed inside OMVs, but hitherto robust methods for targeting heterologous proteins to the OMV surface have been lacking. We have exploited our previously developed hemoglobin protease (Hbp) autotransporter platform for display of heterologous polypeptides at the OMV surface. One, two, or three of the Mycobacterium tuberculosis antigens ESAT6, Ag85B, and Rv2660c were targeted to the surface of Escherichia coli OMVs upon fusion to Hbp. Furthermore, a hypervesiculating ΔtolR ΔtolA derivative of attenuated Salmonella enterica serovar Typhimurium SL3261 was generated, enabling efficient release and purification of OMVs decorated with multiple heterologous antigens, exemplified by the M. tuberculosis antigens and epitopes from Chlamydia trachomatis major outer membrane protein (MOMP). Also, we showed that delivery of Salmonella OMVs displaying Ag85B to antigen-presenting cells in vitro results in processing and presentation of an epitope that is functionally recognized by Ag85B-specific T cell hybridomas. In conclusion, the Hbp platform mediates efficient display of (multiple) heterologous antigens, individually or combined within one molecule, at the surface of OMVs. Detection of antigen- specific immune responses upon vesicle-mediated delivery demonstrated the potential of our system for vaccine development. © 2014, American Society for Microbiology.

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DO - 10.1128/AEM.01941-14

M3 - Article

VL - 80

SP - 5854

EP - 5864

JO - Applied and Environmental Microbiology

T2 - Applied and Environmental Microbiology

JF - Applied and Environmental Microbiology

SN - 0099-2240

IS - 18

ER -