Construction of a recombinant food-grade Lactococcus lactis expressing P23 protein of Cryptosporidium parvum

Liu, Xuehan; Deng, Lei; Li, Wei; Zhong, Zhijun; Zhou, Ziyao; Peng, Guangneng

doi:10.1007/s12223-021-00923-8

Cited by 2 publications

(2 citation statements)

References 28 publications

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“…The washing processes were repeated after incubation (1 h, 37 °C). Ultimately, the membrane was subjected to development using diaminobenzidine tetrahydrochloride (DAB) (Liu et al 2022 ).…”

Section: Methodsmentioning

confidence: 99%

Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Kazemi-Roudsari,

Doosti,

Jami

2024

AMB Expr

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Brucellosis is regarded as one of the world’s most severe zoonotic diseases. This study aimed to investigate the possibility of using recombinant Lactococcus lactis (L. lactis) as a live vector to produce recombinant Brucella abortus (B. abortus) Omp10. The gene sequences were obtained from GenBank. The proteins’ immunogenicity was assessed using Vaxijen. After confirming the cloning of the Omp10 gene in the pNZ8148 vector by enzymatic digestion and PCR, transformation into L. lactis was done. SDS-PAGE and western blot methods evaluated omp10 protein expression. Mice received oral recombinant L. lactis vaccines. IgG antibodies against Omp10 were tested using ELISA. Real-time PCR and ELISA were used to analyze cytokine responses. Survival rate and histopathological changes were evaluated after the challenge. Omp10 was chosen for its 1.5524 antigenicity score. Enzymatic digestion and PCR identified a 381-bp gene fragment. A 10 kDa band indicated the success of L. lactis transformation. Mice administered the L. lactis-pNZ8148-Omp10-Usp45 vaccination 14 days after priming showed significantly higher Omp10-specific total IgG and IgG1 (P < 0.001) than the PBS control group. The mice who received the L. lactis-pNZ8148-Omp10-Usp45 and IRBA vaccines had significantly elevated levels of IFN-γ, TNFα, IL-4, and IL-10 in samples collected on days 14 and 28 (P < 0.001). Inflammatory response, morphological damage, alveolar edema, and lymphocyte infiltration were reduced in the target group. A recombinant L. lactis expressing the Omp10 protein was constructed as an oral Lactococcus-based vaccine and compared to live attenuated vaccines for future brucellosis investigations.

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Section: Methodsmentioning

confidence: 99%

Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Kazemi-Roudsari,

Doosti,

Jami

2024

AMB Expr

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“…The ligation fragment was prepared using seamless clone PreMix (Monad, China) and then electrotransformed into the NZ3900 strain. 22 The vectors containing the ligation fragments were constructed.…”

Section: Construction Of the Recombinant Plasmid And L Lactis Strainmentioning

confidence: 99%

Development of secretory and cell wall anchor recombinant Lactococcus lactis vaccine expressing the porcine enterotoxigenic Escherichia coli K88ac or K99-987P protein

Zhang

Zhao

et al. 2023

Preprint

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Background: Porcine enterotoxigenic Escherichia coli (ETEC) is one of the primary pathogens causing severe diarrhea in piglets. The main pathogenic factors of this strain are K88ac, K99, and 987P proteins. Methods: The K88ac or K99-987P sequence was used to replace the central part of the AcmA gene, and the cell wall anchor-expressing recombinant pNZ8149-MD/Lactobacillus lactisstrain was constructed. The Usp45 signal peptide, dendritic cell-targeting peptide, and TAT sequence were added to the terminal, and the secretory protein-expressing recombinant pNZ8149-XDT /L.lactis strain was constructed. Mice were immunized with the recombinant strains, and mouse K88, K99, 987P, IgG, sIgA antibodies; IFN-γ; and IL-4 levels were quantified. Results: The experimental group showed increased levels of these immune proteins. sIgA level increased drastically, and the levels of K88, K99, 987P, IgG, and IL-4 also increased significantly. Immunized mice were challenged with a strain of ETEC. The XDT feeding group showed the lowest mortality rate, lower symptom score, lower levels of inflammatory cytokines IL-6 and TNF-α, and milder pathological changes, thus adequately protecting the mice from ETEC infection. Limitations: It remains unclear whether the vaccine can protect pigs from infection with ETEC. Conclusion: The XDT feeding vaccine with K88 or K99-987P antigen was developed.

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Construction of a recombinant food-grade Lactococcus lactis expressing P23 protein of Cryptosporidium parvum

Cited by 2 publications

References 28 publications

Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Design of an oral vaccine using Lactococcus lactis against brucellosis: an in vitro and in vivo study

Development of secretory and cell wall anchor recombinant Lactococcus lactis vaccine expressing the porcine enterotoxigenic Escherichia coli K88ac or K99-987P protein

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