Expression of a Porphyromonas gingivalis hemagglutinin on the surface of a Salmonella vaccine vector

Isoda, Ryutaro; Simanski, Scott; Pathangey, Latha B.; Stone, Amy E.S.; Brown, Thomas A.

doi:10.1016/j.vaccine.2006.06.085

Cited by 25 publications

(29 citation statements)

References 31 publications

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“…Overexpression of foreign proteins can also result in mutations in the antigen gene promoter or coding sequence, thus reducing or compromising the desired immune response. Several approaches have been used to address these problems, including truncating the antigen, reducing plasmid copy number, chromosomal expression, reducing promoter activity (33), codon optimization of the antigen sequence (72), or exporting the antigen to an extracellular compartment (25). Another popular approach is the use of in vivo inducible promoters, including pagC (31), nirB (10), and spv and dps promoters (44).…”

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confidence: 99%

SalmonellaVaccine Vectors Displaying Delayed Antigen SynthesisInVivoTo Enhance Immunogenicity

Wang

Scarpellini

et al. 2010

Infect Immun

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We have developed a regulated delayed antigen synthesis (RDAS) system for use in recombinant attenuated Salmonella vaccine (RASV) strains to enhance immune responses by reducing the adverse effects of high-level antigen synthesis. This system includes a chromosomal repressor gene, lacI, expressed from the arabinoseregulated araC P BAD promoter. LacI serves to regulate expression from a plasmid promoter, P trc , that directs antigen synthesis. In the presence of arabinose LacI is produced, which binds to P trc , blocking antigen synthesis. In vivo, an arabinose-poor environment, the concentration of LacI decreases with each cell division, allowing increased antigen synthesis. To optimize the system and for comparison, we altered the lacI ribosomebinding site, start codon, and/or codon content to construct RDAS strains 9095, 9959, and 9241, synthesizing from low to high levels of LacI, respectively, and non-RDAS strain 9555 as a control. We evaluated this system with two test antigens, the green fluorescent protein for initial in vitro assessment and the Streptococcus pneumoniae PspA protein for validation of our system in mice. All RASV strains expressing PspA generated high antilipopolysaccharide antibody titers, indicating that expression of lacI did not interfere with the capacity to induce an immune response. Strain 9241 induced significantly higher anti-PspA IgG and IgA antibody titers than strain 9555, which expressed PspA constitutively. Anti-PspA antibody titers were inversely correlated to the level of LacI synthesis. Strain 9241 also induced significantly greater protective efficacy against challenge with virulent S. pneumoniae. These results suggest that regulated delayed antigen synthesis is useful for improving immunogenicity of RASV strains.

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confidence: 99%

SalmonellaVaccine Vectors Displaying Delayed Antigen SynthesisInVivoTo Enhance Immunogenicity

Wang

Scarpellini

et al. 2010

Infect Immun

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“…The use of plasmid-based expression systems to elevate foreign antigen levels in Salmonella live vaccine vectors is ideal, but these systems have the serious disadvantages of instability and selective requirements in vivo (10,14,17,38). The problem of plasmid instability can be overcome by integrating the genes for the foreign antigens into the Salmonella chromosome, but the level of expression of the antigens from a single locus is usually below the desired levels (33).…”

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confidence: 99%

Oral Vaccination with AttenuatedSalmonella entericaStrains Encoding T-Cell Epitopes from Tumor Antigen NY-ESO-1 Induces Specific Cytotoxic T-Lymphocyte Responses

Meng

Dong

Huang

et al. 2010

Clin Vaccine Immunol

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Bacterial fimbriae can accept foreign peptides and display them on the cell surface. A highly efficient gene replacement method was used to generate peptide vaccines based on Salmonella enterica serovar Typhimurium SL3261. The T-cell epitopes (NY-ESO-1 p157-165 and p157-167) from NY-ESO-1, which is a promising target antigen in patients for the specific immune recognition of cancer, were incorporated into the gene encoding AgfA (the major subunit protein of thin aggregative fimbriae of Salmonella) by replacing an equal length of the DNA segment. To improve cytotoxic T-lymphocyte recognition, both termini of the peptide were flanked by double alanine (AA) residues. Immunofluorescence microscopy with AgfA-specific antiserum verified the expression of chimeric AgfA, which was also proved by a Congo red binding assay. Modulating the immune system to control cancer has been a challenge for cancer immunotherapists for many years. A number of clinical trials have indeed demonstrated that objective tumor regressions can occur in the setting of cancer vaccinations (1,19,31

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“…Lpp-OmpA is an efficient hybrid display system developed by Georgiou et al (9). The system combines the outer membrane-targeting function of E. coli lipoprotein (Lpp) and the transmembrane function of E. coli OmpA and succeeded in anchoring a variety of proteins, including some enzymes, onto the cell surface (10,21). In the present study, according to the successful mode of Lpp-OmpA, two lipoproteins (E. coli Lpp and V. anguillarum Wza) and three V. anguillarum outer membrane proteins (Omporf1, OmpU, and Omp26La) were selected, and their outer membrane targeting regions, indicated by bioinformatics analysis, were separately cloned and combined into six hybrid display systems Lpp/Wza-Omporf1, Lpp/ Wza-OmpU, and Lpp/Wza-Omp26La to mediate proper localization of passenger protein via C-terminal fusion strategy.…”

Section: Discussionmentioning

confidence: 99%

“…Lpp-OmpA is an efficient surface display system developed by Georgiou et al (9), which has been used successfully to anchor a variety of proteins, including some enzymes, onto the cell surface (10,21). This display system allows C-terminal fusion of the passenger proteins and consists of two key anchoring motifs: (i) the signal sequence and the first nine amino acids of E. coli lipoprotein (Lpp), to target the proteins to the inner face of the outer membrane, and (ii) the transmembrane region (amino acids 46 to 159) of E. coli outer membrane protein A (OmpA), to conduct the proteins across the outer membrane.…”

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confidence: 99%

Novel Bacterial Surface Display Systems Based on Outer Membrane Anchoring Elements from the Marine Bacterium Vibrio anguillarum

Zhao

Liu

Wang

et al. 2008

Appl Environ Microbiol

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Surface display of heterologous peptides and proteins such as receptors, antigens, and enzymes on live bacterial cells is of considerable value for various biotechnological and industrial applications. In this study, a series of novel cell surface display systems were examined by using Vibrio anguillarum outer membrane protein and outer membrane lipoprotein as anchoring motifs. These display systems consist of (i) the signal sequence and first 11 N-terminal amino acids of V. anguillarum outer membrane lipoprotein Wza, or the signal sequence and first 9 N-terminal amino acids of the mature major Escherichia coli lipoprotein Lpp, and (ii) transmembrane domains of V. anguillarum outer membrane proteins Omporf1, OmpU, or Omp26La. In order to assay the translocation efficiency of constructed display systems in bacteria, green fluorescent protein (GFP) was inserted to the systems and the results of GFP surface localization confirmed that four of the six surface display systems could successfully display GFP on the E. coli surface. For assaying its potential application in live bacteria carrier vaccines, an excellent display system Wza-Omporf1 was fused with the major capsid protein (MCP) of large yellow croaker iridovirus and introduced into attenuated V. anguillarum strain MVAV6203, and subsequent analysis of MCP surface localization proved that the novel display system Wza-Omporf1 could function as a strong tool in V. anguillarum carrier vaccine development.The display of heterologous proteins or peptides on the surface of prokaryotic or eukaryotic cells, especially bacterial and yeast cells (8), enabled by means of recombinant DNA technology, has become an increasingly used strategy in various applications, including live vaccine development, antibody production, peptide library screening, the use of environmental bioadsorbents, and whole-cell biocatalysis (12). A variety of surface anchoring motifs have been used to establish display systems, including various outer membrane proteins, lipoproteins, autotransporters, subunits of surface appendages, and S-layer proteins (12, 16). The efficiency of surface display systems are highly related to the characteristics of the carrier protein, passenger protein, host cell, and fusion method (17).Lpp-OmpA is an efficient surface display system developed by Georgiou et al. (9), which has been used successfully to anchor a variety of proteins, including some enzymes, onto the cell surface (10, 21). This display system allows C-terminal fusion of the passenger proteins and consists of two key anchoring motifs: (i) the signal sequence and the first nine amino acids of E. coli lipoprotein (Lpp), to target the proteins to the inner face of the outer membrane, and (ii) the transmembrane region (amino acids 46 to 159) of E. coli outer membrane protein A (OmpA), to conduct the proteins across the outer membrane. Since the Lpp-OmpA mode combines the anchoring capabilities of lipoprotein and outer membrane protein, it has outstanding advantages in high surface display efficiency and st...

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Expression of a Porphyromonas gingivalis hemagglutinin on the surface of a Salmonella vaccine vector

Cited by 25 publications

References 31 publications

SalmonellaVaccine Vectors Displaying Delayed Antigen SynthesisInVivoTo Enhance Immunogenicity

SalmonellaVaccine Vectors Displaying Delayed Antigen SynthesisInVivoTo Enhance Immunogenicity

Oral Vaccination with AttenuatedSalmonella entericaStrains Encoding T-Cell Epitopes from Tumor Antigen NY-ESO-1 Induces Specific Cytotoxic T-Lymphocyte Responses

Novel Bacterial Surface Display Systems Based on Outer Membrane Anchoring Elements from the Marine Bacterium Vibrio anguillarum

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