Evaluation of different promoter sequences and antigen sorting signals on the immunogenicity of Bacillus subtilis vaccine vehicles

Paccez, Juliano D.; Nguyen, Hoang Duc; Luiz, Wilson Barros; Ferreira, Rita de Cássia Café; Sbrogio-Almeida, Maria E.; Schuman, Wolfgang; Ferreira, Luís Carlos de Souza

doi:10.1016/j.vaccine.2007.04.021

Cited by 36 publications

(46 citation statements)

References 23 publications

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“…Under our experimental conditions Stx2B is expressed by vegetative cells in amounts lower than previously detected with the ETEC LTB (16,17). The anti-Stx2B sera employed to monitor the antigen in bacterial extracts detected a minimum of 120 ng of Stx2B toxin, while other sera, such as those previously used to quantify LTB, had a higher sensitivity (16,17). Additionally, the low immunogenicity of Stx2B, under different vaccine formulations, has been previously reported (1,2,3,11,12).…”

Section: Discussionmentioning

confidence: 48%

“…This finding may be reasoned by the reduced in vitro expression of Stx2B by recombinant B. subtilis cells and the relatively low sensitivity of the detection method. Under our experimental conditions Stx2B is expressed by vegetative cells in amounts lower than previously detected with the ETEC LTB (16,17). The anti-Stx2B sera employed to monitor the antigen in bacterial extracts detected a minimum of 120 ng of Stx2B toxin, while other sera, such as those previously used to quantify LTB, had a higher sensitivity (16,17).…”

Section: Discussionmentioning

confidence: 58%

“…Bacillus subtilis strains have been engineered to express different antigens as live recombinant vaccine vehicles delivered via the oral route either as spores or vegetative cells (5,6,8,16,17,21). Previous attempts to express the B subunit of heat labile toxin (LTB), produced by enterotoxigenic E. coli (ETEC) strains, have demonstrated that orally delivered B. subtilis vaccine strains may elicit protective systemic antibody responses to a lethal toxin challenge in mice (17).…”

Section: Shiga-like Toxin (Stx)-producing Enterohaemorragicmentioning

confidence: 99%

“…Previous attempts to express the B subunit of heat labile toxin (LTB), produced by enterotoxigenic E. coli (ETEC) strains, have demonstrated that orally delivered B. subtilis vaccine strains may elicit protective systemic antibody responses to a lethal toxin challenge in mice (17). These results suggest that EHEC vaccines based on Stx2B-expressing B. subtilis strains may be feasible.…”

Section: Shiga-like Toxin (Stx)-producing Enterohaemorragicmentioning

confidence: 99%

“…SDS-PAGE and immunoblots of recombinant B. subtilis strains, as well as purified Stx2B, were carried out by previously reported procedures (16,17). Detection of Stx2B in immunoblots was carried out with a mouse anti-Stx2B serum generated in the laboratory with recombinant affinity purified Stx2B produced in E. coli and an anti-mouse IgG peroxidase conjugate (Sigma).…”

Section: Detection Of Stx2b By Sds-page and Immunoblotsmentioning

confidence: 99%

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Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7

Gomes¹,

Bentancor²,

Paccez³

et al. 2009

Braz. J. Microbiol.

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No effective vaccine or immunotherapy is presently available for patients with the hemolytic uremic syndrome (HUS) induced by Shiga-like toxin (Stx) produced by enterohaemorragic Escherichia coli (EHEC) strains, such as those belonging to the O157:H7 serotype. In this work we evaluated the performance of Bacillus subtilis strains, a harmless spore former gram-positive bacterium species, as a vaccine vehicle for the expression of Stx2B subunit (Stx2B). A recombinant B. subtilis vaccine strain expressing Stx2B under the control of a stress inducible promoter was delivered to BALB/c mice via oral, nasal or subcutaneous routes using both vegetative cells and spores. Mice immunized with vegetative cells by the oral route developed low but specific anti-Stx2B serum IgG and fecal IgA responses while mice immunized with recombinant spores developed anti-Stx2B responses only after administration via the parenteral route. Nonetheless, serum anti-Stx2B antibodies raised in mice immunized with the recombinant B. subtilis strain did not inhibit the toxic effects of the native toxin, both under in vitro and in vivo conditions, suggesting that either the quantity or the quality of the induced immune response did not support an effective neutralization of Stx2 produced by EHEC strains.

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Section: Discussionmentioning

confidence: 48%

Section: Discussionmentioning

confidence: 58%

Section: Shiga-like Toxin (Stx)-producing Enterohaemorragicmentioning

confidence: 99%

Section: Shiga-like Toxin (Stx)-producing Enterohaemorragicmentioning

confidence: 99%

Section: Detection Of Stx2b By Sds-page and Immunoblotsmentioning

confidence: 99%

See 3 more Smart Citations

Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7

Gomes¹,

Bentancor²,

Paccez³

et al. 2009

Braz. J. Microbiol.

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Targeting ideal oral vaccine vectors based on probiotics: a systematical view

Jiang

et al. 2019

Appl Microbiol Biotechnol

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Probiotics have great potential to be engineered into oral vaccine delivery systems, which can facilitate elicitation of mucosal immunity without latent risks of pathogenicity. Combined with the progressive understanding of probiotics and the mucosal immune system as well as the advanced biotechniques of genetic engineering, the development of promising oral vaccine vectors based on probiotics is available while complicated and demanding. Therefore, a systematical view on the design of practical probiotic vectors is necessary, which will help to logically analyze and resolve the problems that might be neglected during our exploration. Here, we attempt to systematically summarize several fundamental issues vital to the effectiveness of the vector of probiotics, including the stability of the engineered vectors, the optimization of antigen expression, the improvement of colonization, and the enhancement of immunoreactivity. We also compared the existent strategies and some developing ones, attempting to figure out an optimal strategy that might deserve to be referred in the future development of oral vaccine vectors based on probiotics.

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Bacillus subtilis Endospores at High Purity and Recovery Yields: Optimization of Growth Conditions and Purification Method

et al. 2012

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Bacillus subtilis endospores have applications in different fields including their use as probiotics and antigen delivery vectors. Such specialized applications frequently require highly purified spore preparations. Nonetheless, quantitative data regarding both yields and purity of B. subtilis endospores after application of different growth conditions and purification methods are scarce or poorly reported. In the present study, we conducted several quantitative and qualitative analyses of growth conditions and purification procedures aiming generation of purified B. subtilis spores. Based on two growth media and different incubations conditions, sporulation frequencies up to 74.2 % and spore concentrations up to 7 × 10(9) spores/ml were achieved. Application of a simplified spore isolation method, in which samples were incubated with lysozyme and a detergent, resulted in preparations with highly purified spores at the highest yields. The present study represents, therefore, an important contribution for those working with B. subtilis endospores for different biotechnological purposes.

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Evaluation of different promoter sequences and antigen sorting signals on the immunogenicity of Bacillus subtilis vaccine vehicles

Cited by 36 publications

References 23 publications

Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7

Antibody responses elicited in mice immunized with Bacillus subtilis vaccine strains expressing Stx2B subunit of enterohaemorragic Escherichia coli O157:H7

Targeting ideal oral vaccine vectors based on probiotics: a systematical view

Bacillus subtilis Endospores at High Purity and Recovery Yields: Optimization of Growth Conditions and Purification Method

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