Expression of single-chain antibody against RgpA protease of Porphyromonas gingivalis in Lactobacillus

Marcotte, Harold; Kõll-Klais, Piret; Hultberg, Anna; Zhao, Yaofeng; Gmür, Rudolf; Mändar, Reet; Mikelsaar, Marika; Hammarström, Lennart

doi:10.1111/j.1365-2672.2005.02786.x

Cited by 40 publications

(29 citation statements)

References 25 publications

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“…Correct insertions of the VHH genes into the plasmids were verified by sequencing of the complete expression cassettes. The expression plasmids were transfected into L. paracasei BL23 by electroporation as previously described (23,32), generating strains of Lactobacillus expressing the anti-TcdB VHH antibodies, either secreted or cell wall displayed (see Table S2). …”

Section: Methodsmentioning

confidence: 99%

Neutralization of Clostridium difficile Toxin B Mediated by Engineered Lactobacilli That Produce Single-Domain Antibodies

et al. 2016

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cClostridium difficile is the primary cause of nosocomial antibiotic-associated diarrhea in the Western world. The major virulence factors of C. difficile are two exotoxins, toxin A (TcdA) and toxin B (TcdB), which cause extensive colonic inflammation and epithelial damage manifested by episodes of diarrhea. In this study, we explored the basis for an oral antitoxin strategy based on engineered Lactobacillus strains expressing TcdB-neutralizing antibody fragments in the gastrointestinal tract. Variable domain of heavy chain-only (VHH) antibodies were raised in llamas by immunization with the complete TcdB toxin. Four unique VHH fragments neutralizing TcdB in vitro were isolated. When these VHH fragments were expressed in either secreted or cell wall-anchored form in Lactobacillus paracasei BL23, they were able to neutralize the cytotoxic effect of the toxin in an in vitro cell-based assay. Prophylactic treatment with a combination of two strains of engineered L. paracasei BL23 expressing two neutralizing anti-TcdB VHH fragments (VHH-B2 and VHH-G3) delayed killing in a hamster protection model where the animals were challenged with spores of a TcdA ؊ TcdB ؉ strain of C. difficile (P < 0.05). Half of the hamsters in the treated group survived until the termination of the experiment at day 5 and showed either no damage or limited inflammation of the colonic mucosa despite having been colonized with C. difficile for up to 4 days. The protective effect in the hamster model suggests that the strategy could be explored as a supplement to existing therapies for patients.

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Section: Methodsmentioning

confidence: 99%

Neutralization of Clostridium difficile Toxin B Mediated by Engineered Lactobacilli That Produce Single-Domain Antibodies

et al. 2016

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“…Lactobacilli are not existing usually in sulcus of gingiva but they can be formulated to show its antibacterial property on P gingivalis. Such a kind of method, organized by means of the innate immunomodulatory properties of probiotics, influence deal remarkable predictions in the coming years [13] .…”

Section: Anticipated Mechanisms Of Probiotic Activity With Respect Tomentioning

confidence: 99%

Probiotic significance in periodontics- A short review

Shetty¹,

Bhandary²,

Thomas³

et al. 2017

Int J Dentistry Res

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Background: Elimination of periodontal pathogen is the main aim of during a periodontal therapy, and scaling root planning is the gold standard. Use of antibiotic for long duration is not advisable due to increased bacterial resistance. Thus adjunctive therapy comes into importance for maintaining good oral flora. Use of probiotic has shown improvement of microbial flora of the gut thus helping to maintain a healthy living. Studies and research on the effect of probiotic on oral microflora will ensure future of periodontal therapy. Aim: To assess the clinical efficacy of probiotics in periodontal health. Material and Methods: A literature survey was conducted by collecting research studies using keywords like "Probiotics, periodontal health, application" using search engines like google search, pubmed. Result: Supporting studies were selected which shows that probiotic is a beneficial formula for improving oral health Conclusion: Earlier work has proved that when probiotics used in a certain amount helps in improving the oral flora which proves to be a promising tool in the treatment of periodontal diseases.

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“…In each cassette, the antibody fragment was also fused to an E tag for detection with an anti-E-tag antibody in Western blotting and enzyme-linked immunosorbent assay (ELISA). The pAF plasmid series (pAF100 to pAF1100) containing the 11 expression cassettes was introduced into L. paracasei (previously known as L. casei or L. zeae ATCC 393 pLZ15 Ϫ ) (7,15) by electroporation as previously described (7,10), generating L. paracasei pAF100 to pAF1100. The most important bacterial strains and plasmids used in this study are listed in Table 1.…”

mentioning

confidence: 99%

Integrative Expression System for Delivery of Antibody Fragments by Lactobacilli

Martín

Pant

Ladero

et al. 2011

Appl Environ Microbiol

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Expression of single-chain antibody against RgpA protease of Porphyromonas gingivalis in Lactobacillus

Cited by 40 publications

References 25 publications

Neutralization of Clostridium difficile Toxin B Mediated by Engineered Lactobacilli That Produce Single-Domain Antibodies

Neutralization of Clostridium difficile Toxin B Mediated by Engineered Lactobacilli That Produce Single-Domain Antibodies

Probiotic significance in periodontics- A short review

Integrative Expression System for Delivery of Antibody Fragments by Lactobacilli

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