• Journal of Microbiology and Biotechnology
• /
• v.27 no.8
• /
• pp.1519-1528
• /
• 2017

Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.12
• /
• pp.2095-2105
• /
• 2018

In our previous studies, we have identified several in vivo-induced antigens and evaluated their potential as subunit vaccine candidates in a murine model, in which the recombinant protein GalT showed the most potent immunogenicity and immunoprotective efficacy against Actinobacillus pleuropneumoniae. To exploit a more efficient way of delivering GalT proteins, in this study, we employed the widely studied E. coli outer membrane vesicles (OMVs) as a platform to deliver GalT protein and performed the vaccine trial using the recombinant GalT-OMVs in the murine model. Results revealed that GalT-OMVs could elicit a highly-specific, IgG antibody titer that was comparable with the adjuvant GalT group. Significantly higher lymphocyte proliferation and cytokines secretion levels were observed in the GalT-OMVs group. 87.5% and 50% of mice were protected from a lethal dose challenge using A. pleuropneumoniae in active or passive immunization, respectively. Histopathologic and immunohistochemical analyses showed remarkably reduced pathological changes and infiltration of neutrophils in the lungs of mice immunized with GalT-OMVs after the challenge. Taken together, these findings confirm that OMVs can be used as a platform to deliver GalT protein and enhance its immunogenicity to induce both humoral and cellular immune responses in mice.

• Journal of Microbiology and Biotechnology
• /
• v.32 no.5
• /
• pp.621-629
• /
• 2022

Bacterial outer membrane vesicles (OMVs) typically contain multiple immunogenic molecules that include antigenic proteins, making them good candidates for vaccine development. In animal models, vaccination with OMVs has been shown to confer protective immune responses against many bacterial diseases. It is possible to genetically introduce heterologous protein antigens to the bacterial host that can then be produced and relocated to reside within the OMVs by means of the host secretion mechanisms. Accordingly, in this study we sought to develop a novel platform for recombinant OMV (rOMV) production in the widely used bacterial expression host species, Escherichia coli. Three different lipoprotein signal peptides including their Lol signals and tether sequences-from Neisseria meningitidis fHbp, Leptospira interrogans LipL32, and Campylobactor jejuni JlpA-were combined upstream to the GFPmut2 model protein, resulting in three recombinant plasmids. Pilot expression studies showed that the fusion between fHbp and GFPmut2 was the only promising construct; therefore, we used this construct for large-scale expression. After inducing recombinant protein expression, the nanovesicles were harvested from cell-free culture media by ultrafiltration and ultracentrifugation. Transmission electron microscopy demonstrated that the obtained rOMVs were closed, circular single-membrane particles, 20-200 nm in size. Western blotting confirmed the presence of GFPmut2 in the isolated vesicles. Collectively, although this is a non-optimized, proof-of-concept study, it demonstrates the feasibility of this platform in directing target proteins into the vesicles for OMV-based vaccine development.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.8
• /
• pp.1343-1347
• /
• 2016

Outer membrane vesicles (OMVs) are spherical nanostructures that are ubiquitously shed from gram-negative bacteria both in vitro and in vivo. Recent findings revealed that OMVs, which contain diverse components derived from the parent bacterium, play an important role in communication with neighboring bacteria and the environment. Furthermore, nanoscale proteoliposomes decorated with pathogen-associated molecules attract considerable attention as a non-replicative carrier for vaccines and drug materials. This review introduces recent advances in OMV biogenesis and discusses the roles of OMVs in the context of bacterial communication and virulence regulation. It also describes the remarkable accomplishments in OMV engineering for diverse therapeutic applications.

• Journal of Food Hygiene and Safety
• /
• v.16 no.4
• /
• pp.333-341
• /
• 2001

The actual overall migration data obtained from plastic food packaging materials into food simulants under high temperature testing conditions as described in the regulations of European Union, USA, and Korea or Japan were compared. Overall migration values(OMVs) with non-fatty food simulants under high temperature conditions were observed to be generally below 2.5 mg/dm$^2$ except polyamides(CPA and PA 6,6) which were tested at 121$^{\circ}C$ for 2 hrs. As for the fatty food simulants, the OMVs with soybean oil were higher than other simulants. Among the films tested, PVC wrap showed higher OMVs ranging between 23.9 and 54.6 mg/dm$^2$than others. The OMVs were measured at higher level with the elevation of contact temperature and the extension of contact time, and in fatty food simulants rather than in non-fatty simulants. Under similar testing temperature and time conditions. the OMVs tended to be increased in polar films like PA with polar simulants, and contrarily in non-polar films like PO with non-polar simulants. It is noteworthy that a discrepancy with regard to the result of OMVs was observed for some films as a result of different migration testing methods and conditions of each country areas.

• Journal of Life Science
• /
• v.28 no.4
• /
• pp.496-507
• /
• 2018

Immunization has been performed for centuries and is generally accepted as a sustainable method of controlling bacteria, viruses, and mediated and infectious diseases. Despite many studies having been performed on animal subjects to demonstrate the importance of toxin immunity, the use of toxoid vaccines in humans and animals has been limited for a long time. Recently, the development of the toxoid antigen delivery system has been facilitated using novel nano-medicinal technology. The micro/nano-carrier has been used to improve vaccination coverage as well as reduce vaccine costs. A micro/nano-carrier is a micro/nano-sized material that delivers immune cargo, including recombinant or peptide toxoid antigens. These toxoid antigens are either encapsulated in the interior or displayed on the surface of micro/nano-carriers as a way to protect them from the cellular machinery. In particular, the combination of toxoid antigens and micro/nano-carriers can induce phagocytosis through the specific interactions between GCs and macrophages; thus, the toxoid antigens can be delivered easily into the macrophages. This paper reviews recent achievements of micro/nano-carriers in the field of vaccine delivery systems such as microbial ghost cells (GCs, Bacterial ghost cells and Yeast ghost cells), gene-manipulated outer membrane vesicles (OMVs) and biocompatible, polymer-based nanoparticles (NPs, NP-Carrier and NP-Cage). Finally, this review shows various aspects in terms of the hosts' immune responses.

• Journal of Microbiology and Biotechnology
• /
• v.19 no.10
• /
• pp.1271-1279
• /
• 2009

To develop low endotoxic and multi-immunogenic outer membrane vesicles (OMVs), a deletion mutant of the msbB gene in Salmonella enterica serovar Typhimurium (S. Typhimurium) was used as a source of low endotoxic OMV, and an expression vector of the canine parvovirus (CPV) VP2 epitope fused to the bacterial OmpA protein was constructed and transformed into the Salmonella ${\Delta}msbB$ mutant. In a lethality test, BALB/c mice injected intraperitoneally with the Salmonella ${\Delta}msbB$ mutant survived for 7 days, whereas mice injected intraperitoneally with the wild type survived for 3 days. Moreover, all mice inoculated orally with the ${\Delta}msbB$ mutant survived for 30 days, but 80% of mice inoculated orally with the wild type survived. The OmpA::CPV VP2 epitope fusion protein was expressed successfully and associated with the outer membrane and OMV fractions from the mutant S. Typhimurium transformed with the fusion protein-expressing vector. In immunogenicity tests, sera obtained from the mice immunized with either the Salmonella msbB mutant or its OMVs containing the OmpA::CPV VP2 epitope showed bactericidal activities against wild-type S. Typhimurium and contained specific antibodies to the CPV VP2 epitope. In the hemagglutination inhibition (HI) assay as a measurement of CPV-neutralizing activity in the immune sera, there was an 8-fold increase of HI titer in the OMV-immunized group compared with the control. These results suggested that the CPV-neutralizing antibody response was raised by immunization with OMV containing the OmpA::CPV VP2 epitope, as well as the protective immune response against S. Typhimurium in BALB/c mice.

• Translational and Clinical Pharmacology
• /
• v.26 no.3
• /
• pp.103-110
• /
• 2018

The human microbiome is known to play an essential role in influencing host health. Extracellular vesicles (EVs) have also been reported to act on a variety of signaling pathways, distally transport cellular components such as proteins, lipids, and nucleic acid, and have immunomodulatory effects. Here we shall review the current understanding of the intersectionality of the human microbiome and EVs in the emerging field of microbiota-derived EVs and their pharmacological potential. Microbes secrete several classes of EVs: outer membrane vesicles (OMVs), membrane vesicles (MVs), and apoptotic bodies. EV biogenesis is unique to each cell and regulated by sophisticated signaling pathways. EVs are primarily composed of lipids, proteins, nucleic acids, and recent evidence suggests they may also carry metabolites. These components interact with host cells and control various cellular processes by transferring their constituents. The pharmacological potential of microbiome-derived EVs as vaccine candidates, biomarkers, and a smart drug delivery system is a promising area of future research. Therefore, it is necessary to elucidate in detail the mechanisms of microbiome-derived EV action in host health in a multi-disciplinary manner.

• KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
• /
• v.24 no.3
• /
• pp.97-106
• /
• 2018

This study aimed at examining the suitability of $Tenax^{(R)}$ for the migration testing of food packaging materials, which is currently approved in the EU as a dry food simulant. The results are used as a basis to examine the feasibility of introducing $Tenax^{(R)}$ to Korean regulation. The OMVs of test specimen into various solvents (diethyl ether, ethanol, pentane, and acetone) after exposure to $100^{\circ}C$ for 1 hr were compared. Diethyl ether showed the highest OMV ($1.33mg/dm^2$) among the solvents tested. When the tests were conducted with different amounts of $Tenax^{(R)}$ of 2, 4, or 8 g per specimen, the OMVs were 0.75, 1.33 and $1.40mg/dm^2$, respectively. The OMV obtained with a closed system after wrapping with aluminum foil showed a significantly higher OMV ($1.61mg/dm^2$) than that without aluminum wrapping ($1.318mg/dm^2w$) and an open system without lid ($1.06mg/dm^2$). The specific migration rates of surrogates spiked in the polyethylene test film and paper samples into $Tenax^{(R)}$ were compared with those into liquid food simulants including 95% ethanol and n-heptane, and actual foods such as starch, skim milk, and sugar. In general, the specific migration levels of surrogates into $Tenax^{(R)}$ were similar compared with n-heptane, however those were significantly higher than into actual foods. These results suggest that $Tenax^{(R)}$ may be used as a food simulant for the long-term preservation of dried foods and paper products. However, more studies need to be conducted to investigate the factors influencing the migration into $Tenax^{(R)}$, such as the types of foods and packaging materials tested, migration conditions, and surrogates properties etc.