Expression of Helicobacter pylori CagL gene in Lactococcus lactis MG1363 and evaluation of its immunogenicity as an oral vaccine in mice

Aliramaei, Mohammad Reza; Khorasgani, Mohammad Rabbani; Rahmani, Mohammad Reza; Zarkesh-Esfahani, Hamid; Emamzadeh, Rahman

doi:10.1016/j.micpath.2019.103926

Cited by 17 publications

(15 citation statements)

References 38 publications

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“…As happened with the aforementioned replicons, the majority of the studies with pAMβ1-based vectors address the use of L. lactis for live mucosal vaccination, with antigens being expressed intracellularly, anchored at the cell surface or secreted to the host mucosa. Aliramaei et al [ 179 ] cloned the H. pylori cagL gene, which expresses a highly conserved protein located at the tip of a pili, into the pAMJ2008 vector. Although pAMJ2008 is a secretory vector, the expression of the protein was only detected intracellularly.…”

Section: Plasmid Vectors Available For Plasmid Dna and Recombinantmentioning

confidence: 99%

Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens by Lactococcus lactis Cell Factories

Duarte

Monteiro

2021

IJMS

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The Lactococcus lactis bacterium found in different natural environments is traditionally associated with the fermented food industry. But recently, its applications have been spreading to the pharmaceutical industry, which has exploited its probiotic characteristics and is moving towards its use as cell factories for the production of added-value recombinant proteins and plasmid DNA (pDNA) for DNA vaccination, as a safer and industrially profitable alternative to the traditional Escherichia coli host. Additionally, due to its food-grade and generally recognized safe status, there have been an increasing number of studies about its use in live mucosal vaccination. In this review, we critically systematize the plasmid replicons available for the production of pharmaceutical-grade pDNA and recombinant proteins by L. lactis. A plasmid vector is an easily customized component when the goal is to engineer bacteria in order to produce a heterologous compound in industrially significant amounts, as an alternative to genomic DNA modifications. The additional burden to the cell depends on plasmid copy number and on the expression level, targeting location and type of protein expressed. For live mucosal vaccination applications, besides the presence of the necessary regulatory sequences, it is imperative that cells produce the antigen of interest in sufficient yields. The cell wall anchored antigens had shown more promising results in live mucosal vaccination studies, when compared with intracellular or secreted antigens. On the other side, engineering L. lactis to express membrane proteins, especially if they have a eukaryotic background, increases the overall cellular burden. The different alternative replicons for live mucosal vaccination, using L. lactis as the DNA vaccine carrier or the antigen producer, are critically reviewed, as a starting platform to choose or engineer the best vector for each application.

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Section: Plasmid Vectors Available For Plasmid Dna and Recombinantmentioning

confidence: 99%

Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens by Lactococcus lactis Cell Factories

Duarte

Monteiro

2021

IJMS

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“…CagL‐specific IgA was also detected in the faeces. This vaccine is therefore immunogenic and able to induce mucosal responses 37 . Mice orally immunised with recombinant L lactis expressing H pylori Lpp20 antigen 38 also had elevated serum IgG levels and reduced gastric urease activity following H pylori challenges.…”

Section: Vaccinesmentioning

confidence: 99%

Review ‐ Helicobacter, inflammation, immunology and vaccines

Robinson

Lehours

2020

Helicobacter

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Understanding the mechanisms involved in induction and regulation of the immune and inflammatory response to Helicobacter pylori is extremely important in determining disease outcomes. H pylori expresses a plethora of factors that influence the host response. Vaccines against H pylori are desperately needed for the prevention of gastric carcinogenesis, especially with the increasing trends in antimicrobial resistance. This review summarizes some important findings, published between 1 April 2019 and 31 March 2020, in the areas of H pylori‐mediated inflammation, immunity and vaccines.

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“…Western blot analysis was used to determine the level of expression, as described previously. 20 After incubation overnight at 4°C and washing three times using sterile PBS buffer, the membrane was incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG (diluted 1:1000; Abcam) at room temperature for 1 h and then reacted with ECL chemiluminescent reagent (Bio-Rad) under dark conditions for 5 min. Finally, the signal was obtained using a ChemiDoc XRS System (Bio-Rad).…”

Section: Western Blot Analysismentioning

confidence: 99%

“…Therefore, the development and evaluation of potential vaccines to prevent and halt the spread of H. pylori is of high priority. Currently, a range of approaches to the design of an H. pylori vaccine have been developed in preclinical or clinical trials, 7 including dendritic cell‐based vaccines, 8 subunit vaccines, 9–19 bacterial vector vaccines, 20–31 and DNA vaccines 32–34 . Of note, adjuvanted H. pylori vaccines can induce significant immune protection against H. pylori infection 35–42 .…”

Section: Introductionmentioning

confidence: 99%

“…a range of approaches to the design of an H. pylori vaccine have been developed in preclinical or clinical trials, 7 including dendritic cellbased vaccines, 8 subunit vaccines, [9][10][11][12][13][14][15][16][17][18][19] bacterial vector vaccines, [20][21][22][23][24][25][26][27][28][29][30][31] and DNA vaccines. [32][33][34] Of note, adjuvanted H. pylori vaccines can induce significant immune protection against H. pylori infection.…”

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Immune evaluation of aSaccharomyces cerevisiae‐based oral vaccine againstHelicobacter pyloriin mice

et al. 2020

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Background Helicobacter pylori (H. pylori) is a common human pathogenic bacterium that is associated with gastric diseases. The current leading clinical therapy is combination antibiotics, but this treatment has safety issues, especially the induction of drug resistance. Therefore, developing a safe and effective vaccine against H. pylori is one of the best alternatives. Objective To develop Saccharomyces cerevisiae (S. cerevisiae)‐based oral vaccines and then demonstrate the feasibility of this platform for preventing H. pylori infection in the absence of a mucosal adjuvant. Materials and Methods Saccharomyces cerevisiae (S. cerevisiae)‐based oral vaccines, including EBY100/pYD1‐UreB and EBY100/pYD1‐VacA, were generated and analyzed by Western blot, Immunofluorescence analysis, flow cytometric assay, and indirect enzyme‐link immunosorbent assay (ELISA). Further, antibody responses induced by oral administration of EBY100/pYD1‐UreB, EBY100/pYD1‐VacA, or EBY100/pYD1‐UreB + EBY100/pYD1‐VacA were measured in a mouse model. Lastly, the vaccinated mice were infected with H. pylori SS1, and colonization in the stomach were evaluated. Results Saccharomyces cerevisiae‐based H. pylori oral vaccines were successfully constructed. Mice orally administered with EBY100/pYD1‐UreB, EBY100/pYD1‐VacA, or EBY100/pYD1‐UreB + EBY100/pYD1‐VacA exhibited a significant humoral immune response as well as a mucosal immune response. Importantly, S. cerevisiae‐based oral vaccines could effectively reduce bacterial loads with statistical significance after H. pylori infection. Conclusions Our study shows that S. cerevisiae‐based platforms can serve as an alternative approach for the future development of promising bacterial oral vaccine candidates.

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Expression of Helicobacter pylori CagL gene in Lactococcus lactis MG1363 and evaluation of its immunogenicity as an oral vaccine in mice

Cited by 17 publications

References 38 publications

Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens by Lactococcus lactis Cell Factories

Plasmid Replicons for the Production of Pharmaceutical-Grade pDNA, Proteins and Antigens by Lactococcus lactis Cell Factories

Review ‐ Helicobacter, inflammation, immunology and vaccines

Immune evaluation of aSaccharomyces cerevisiae‐based oral vaccine againstHelicobacter pyloriin mice

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