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Improved cell surfa...
Improved cell surface display of Salmonella enterica serovar Enteritidis antigens in Escherichia coli
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- Gustavsson, Martin (författare)
- KTH,Industriell bioteknologi
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Do, T. -H (författare)
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- Lüthje, P. (författare)
- Karolinska Institutet
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Tran, N. T. (författare)
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- Brauner, A. (författare)
- Karolinska Institutet
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- Samuelson, Patrik (författare)
- KTH,Industriell bioteknologi
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Truong, N. H. (författare)
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- Larsson, Gen (författare)
- KTH,Industriell bioteknologi
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(creator_code:org_t)
- 2015-04-09
- 2015
- Engelska.
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Ingår i: Microbial Cell Factories. - : Springer Science and Business Media LLC. - 1475-2859. ; 14:1
- Relaterad länk:
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https://microbialcel...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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http://kipublication...
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Abstract
Ämnesord
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- Background: Salmonella enterica serovar Enteritidis (SE) is one of the most potent pathogenic Salmonella serotypes causing food-borne diseases in humans. We have previously reported the use of the β-autotransporter AIDA-I to express the Salmonella flagellar protein H:gm and the SE serotype-specific fimbrial protein SefA at the surface of E. coli as live bacterial vaccine vehicles. While SefA was successfully displayed at the cell surface, virtually no full-length H:gm was exposed to the medium due to extensive proteolytic cleavage of the N-terminal region. In the present study, we addressed this issue by expressing a truncated H:gm variant (H:gmd) covering only the serotype-specific central region. This protein was also expressed in fusion to SefA (H:gmdSefA) to understand if the excellent translocation properties of SefA could be used to enhance the secretion and immunogenicity. Results: H:gmd and H:gmdSefA were both successfully translocated to the E. coli outer membrane as full-length proteins using the AIDA-I system. Whole-cell flow cytometric analysis confirmed that both antigens were displayed and accessible from the extracellular environment. In contrast to H:gm, the H:gmd protein was not only expressed as full-length protein, but it also seemed to promote the display of the protein fusion H:gmdSefA. Moreover, the epitopes appeared to be recognized by HT-29 intestinal cells, as measured by induction of the pro-inflammatory interleukin 8. Conclusions: We believe this study to be an important step towards a live bacterial vaccine against Salmonella due to the central role of the flagellar antigen H:gm and SefA in Salmonella infections and the corresponding immune responses against Salmonella.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
Nyckelord
- AIDA-I
- Autotransport
- Escherichia coli
- Live vaccines
- Salmonella enterica
- Surface expression
- Bacteria (microorganisms)
- Salmonella
- Salmonella enteritidis
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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