Genetically modified Bifidobacterium displaying Salmonella-antigen protects mice from lethal challenge of Salmonella Typhimurium in a murine typhoid fever model

Yamamoto, Sakura; Wada, Jun; Katayama, Takane; Jikimoto, Takumi; Nakamura, M.; Kinoshita, Shohiro; Lee, Kyung Mi; Kawabata, Masato; Shirakawa, Toshiro

doi:10.1016/j.vaccine.2010.08.007

Cited by 30 publications

(17 citation statements)

References 39 publications

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“…These include the SPs of the galacto-N-biose/lacto-N-biose I-binding protein [44] and exo-xylanase [45–47] of B . longum , the β-galactosidase of B .…”

Section: Discussionmentioning

confidence: 99%

A Phytase-Based Reporter System for Identification of Functional Secretion Signals in Bifidobacteria

et al. 2015

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Health-promoting effects have been attributed to a number of Bifidobacterium sp. strains. These effects as well as the ability to colonise the host depend on secreted proteins. Moreover, rational design of protein secretion systems bears the potential for the generation of novel probiotic bifidobacteria with improved health-promoting or therapeutic properties. To date, there is only very limited data on secretion signals of bifidobacteria available. Using in silico analysis, we demonstrate that all bifidobacteria encode the major components of Sec-dependent secretion machineries but only B. longum strains harbour Tat protein translocation systems. A reporter plasmid for secretion signals in bifidobacteria was established by fusing the coding sequence of the signal peptide of a sialidase of Bifidobacterium bifidum S17 to the phytase gene appA of E. coli. The recombinant strain showed increased phytase activity in spent culture supernatants and reduced phytase levels in crude extracts compared to the control indicating efficient phytase secretion. The reporter plasmid was used to screen seven predicted signal peptides in B. bifidum S17 and B. longum E18. The tested signal peptides differed substantially in their efficacy to mediate protein secretion in different host strains. An efficient signal peptide was used for expression and secretion of a therapeutically relevant protein in B. bifidum S17. Expression of a secreted cytosine deaminase led to a 100-fold reduced sensitivity of B. bifidum S17 to 5-fluorocytosine compared to the non-secreted cytosine deaminase suggesting efficient conversion of 5-fluorocytosine to the cytotoxic cancer drug 5-fluorouracil by cytosine deaminase occurred outside the bacterial cell. Selection of appropriate signal peptides for defined protein secretion might improve therapeutic efficacy as well as probiotic properties of bifidobacteria.

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“…These include the SPs of the galacto-N-biose/lacto-N-biose I-binding protein [44] and exo-xylanase [45–47] of B . longum , the β-galactosidase of B .…”

Section: Discussionmentioning

confidence: 99%

A Phytase-Based Reporter System for Identification of Functional Secretion Signals in Bifidobacteria

et al. 2015

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“…pTB6 harbors four open reading frames, including a gene encoding a RepB protein, suggesting a replication by the rolling circle mechanism (82). This replicon has been successfully used in B. longum, B. breve, and B. animalis for expression of E. coli cytosine deaminase or the Salmonella FliC protein (26,27,53,80,87). The replicon of plasmid pMB1 was shown to replicate stably in B. longum, B. animalis, and B. adolescentis by the theta mechanism and was used to express endostatin, tumor necrosis factor (TNF)-related apoptosisinducing ligand, and human interleukin 10 (19, 28, 41, 68, 71, 86).…”

Section: Plasmidsmentioning

confidence: 99%

“…The fusion protein was successfully expressed, as shown by Western blotting and fluorescence microscopy. However, correct location of the fusion protein on the surface was not verified by testing different fractions, e.g., cytoplasm membrane and cell wall, of the recombinant strain (87).…”

Section: Localization Signalsmentioning

confidence: 99%

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Accessing the Inaccessible: Molecular Tools for Bifidobacteria

Sun

Baur

Zhurina

et al. 2012

Appl Environ Microbiol

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ABSTRACTBifidobacteria are an important group of the human intestinal microbiota that have been shown to exert a number of beneficial probiotic effects on the health status of their host. Due to these effects, bifidobacteria have attracted strong interest in health care and food industries for probiotic applications and several species are listed as so-called “generally recognized as safe” (GRAS) microorganisms. Moreover, recent studies have pointed out their potential as an alternative or supplementary strategy in tumor therapy or as live vaccines. In order to study the mechanisms by which these organisms exert their beneficial effects and to generate recombinant strains that can be used as drug delivery vectors or live vaccines, appropriate molecular tools are indispensable. This review provides an overview of the currently available methods and tools to generate recombinant strains of bifidobacteria. The currently used protocols for transformation of bifidobacteria, as well as replicons, selection markers, and determinants of expression, will be summarized. We will further discuss promoters, terminators, and localization signals that have been used for successful generation of expression vectors.

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“…BI-LTB appears to be one of the best forms of LTB adjuvant delivery system [9]. Because of the same reason, the several strains LAB has been genetically modified as a promising oral recombinant vaccine delivery system for inducing efficient mucosal immunity as well as systematic immunity [9,[22][23][24][25][26].…”

Section: Discussionmentioning

confidence: 99%

Oral Administration Recombinant Bifidobacterium-LTB (B Subunit of Heat-Labile Enterotoxin) Enhances the OVA (Ovalbumin)-Specific sIgA in Jejunal Mucosa of Sprague-Dawley (SD) Rat

Ma¹,

Hao²,

Tang³

et al. 2012

IJCM

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The LTB of enterotoxigenic Escherichia coli (ETEC) expressed in Bifidobacterium infantis (BI) has been testified as mucosal adjuvant with co-vaccination BI-CfaB (the major fimbrial subunit) together in vivo in our previous study. In order to investigate the mucosal adjuvant effect of BI-LTB to purified antigens, we oral vaccinated SD rats with recombinant BI-LTB plus OVA (rBI-LTB + OVA), and wild type BI plus OVA (wBI + OVA), OVA and PBS (Phosphate buffered saline) were vaccinated as controls, respectively. The OVA-specific sIgA in jejunal mucosa and specific IgG in serium were measured with ELISA (Enzyme-linked immunosorbent assay) and the sIgA producing cells were detected with immunohistochemistry technology (IHC) and Qwin image manipulation tools subsequently. The results shown rBI-LTB could stimulate SD rats produce high titer OVA-specific sIgA in rBI-LTB + OVA group and the OVA-specific sIgA titer in rBI-LTB + OVA group was found significant greater than that of the wBI + OVA group or OVA single group (p < 0.05). However no such significant difference was detected between the group wBI + OVA and OVA. IHC results suggested that intestinal mucosa and submucosa was the main field of sIgA secretion. These results suggested that recombinant LTB expression in BI could be used as a wide range mucosal adjuvant with different form antigens.

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Genetically modified Bifidobacterium displaying Salmonella-antigen protects mice from lethal challenge of Salmonella Typhimurium in a murine typhoid fever model

Cited by 30 publications

References 39 publications

A Phytase-Based Reporter System for Identification of Functional Secretion Signals in Bifidobacteria

A Phytase-Based Reporter System for Identification of Functional Secretion Signals in Bifidobacteria

Accessing the Inaccessible: Molecular Tools for Bifidobacteria

Oral Administration Recombinant Bifidobacterium-LTB (B Subunit of Heat-Labile Enterotoxin) Enhances the OVA (Ovalbumin)-Specific sIgA in Jejunal Mucosa of Sprague-Dawley (SD) Rat

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