Heterologous expression of antigenic peptides in Bacillus subtilis biofilms

Vogt, Cédric M.; Schraner, Elisabeth M.; Aguilar, Claudio; Eichwald, Catherine

doi:10.1186/s12934-016-0532-5

Cited by 24 publications

(25 citation statements)

References 45 publications

(57 reference statements)

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“…Interestingly, the expression of B. subtilis tasA sinR lux TasA-mCherry was already significantly increased at 24 h postinoculation when it was incubated at 37°C rather than at 30°C. Consistent with the findings of our previous research (13), the sporulation ability of the B. subtilis tasA sinR lux TasA-mCherry strain was decreased compared to that of wt strain B. subtilis lux when it was grown at either 30°C or 37°C (Fig. 2C).…”

Section: Figsupporting

confidence: 92%

“…For this purpose, we incorporated the luxCDABE operon from Photorhabdus luminescens (25) driven by the tapA promoter into a B. subtilis tasA sinR strain also harboring the tapA-sipW-tasA-mCherry operon (Table 1), generating the B. subtilis tasA sinR lux TasA-mCherry strain. The tasA sinR genetic background in B. subtilis improves the expression of heterologous genes driven by the tapA promoter (13). We hypothesized that this newly engineered strain would permit the easy tracking of B. subtilis by luminescence when it is in a vegetative state.…”

Section: Resultsmentioning

confidence: 99%

“…Next, we wondered whether a specific humoral immune response against the integumental antigens from the parasite E. granulosus (EgTrp and EgA31) could be elicited by using the immunization methodology described above. For this purpose, we used two different recombinant B. subtilis spores engineered to express the fusion proteins TasA plus EgTrp from residues 102 to 207 [TasA-(102-207)EgTrp] or TasA plus EgA31 from residues 370 to 583 [TasA-(370-583)EgA31] ( Table 1) (13). For simplification, we refer to these as B. subtilis TasA-EgTrp and B. subtilis TasA-EgA31, respectively.…”

Section: Figmentioning

confidence: 99%

“…For simplification, we refer to these as B. subtilis TasA-EgTrp and B. subtilis TasA-EgA31, respectively. As described by Vogt et al (13), these strains can display the E. granulosus antigenic peptides (EgTrp and EgA31) fused in frame to the C terminus of TasA on the surface of in vitro biofilms. For this experiment, four groups of two dogs each (Table 2) were employed.…”

Section: Figmentioning

confidence: 99%

“…We recently engineered B. subtilis for the display of heterologous proteins on the surface of the biofilm. To this end, the matrix protein TasA was fused to various peptides via its C terminus, and the fusions were expressed under biofilm-inducing conditions (13). To optimize expression, we used the tasA sinR genetic background.…”

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Oral Application of Recombinant Bacillus subtilis Spores to Dogs Results in a Humoral Response against Specific Echinococcus granulosus Paramyosin and Tropomyosin Antigens

et al. 2018

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Bacillus subtilis is known as an endospore-and biofilm-forming bacterium with probiotic properties. We have recently developed a method for displaying heterologous proteins on the surface of B. subtilis biofilms by introducing the coding sequences of the protein of interest into the bacterial genome to generate a fusion protein linked to the C-terminus of the biofilm matrix protein TasA. Although B. subtilis is a regular component of the gut microflora, we constructed a series of recombinant B. subtilis strains that were tested for their ability to immunize dogs following oral application of the spores. Specifically, we tested recombinant spores of B. subtilis carrying either the fluorescent protein mCherry or else selected antigenic peptides (tropomyosin and paramyosin) from Echinococcus granulosus, a zoonotic intestinal tapeworm of dogs and other carnivores. The application of the recombinant B. subtilis spores led to the colonization of the gut with recombinant B. subtilis but did not cause any adverse effect on the health of the animals. As measured by ELISA and immunoblot, the dogs were able to develop a humoral immune response against mCherry as well as against E. granulosus antigenic peptides. Interestingly, the sera of dogs obtained from immunization with recombinant spores of E. granulosus peptides were able to recognize E. granulosus protoscoleces, which represent the infective form of the head of the tapeworms. These results represent an essential step towards the establishment of B. subtilis as an enteric vaccine agent. The application of the recombinant B. subtilis spores led to the colonization of the gut 32 with recombinant B. subtilis but did not cause any adverse effect on the health of the 33 animals. As measured by ELISA and immunoblot, the dogs were able to develop a 34 humoral immune response against mCherry as well as against E. granulosus antigenic 35 peptides. Interestingly, the sera of dogs obtained from immunization with recombinant 36 spores of E. granulosus peptides were able to recognize E. granulosus protoscoleces, 37 which represent the infective form of the head of the tapeworms. These results represent 38 an essential step towards the establishment of B. subtilis as an enteric vaccine agent. 39 40

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Section: Figsupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

mentioning

confidence: 99%

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