Characterization of Outer Membrane Vesicles from<i>Brucella melitensis</i>and Protection Induced in Mice

Ávila-Calderón, Eric Daniel; López-Merino, Ahidé; Jain, Neeta; Peralta, Humberto; López-Villegas, Edgar Óliver; Sriranganathan, Nammalwar; Boyle, Stephen M.; Witonsky, Sharon G.; Contreras-Rodríguez, Araceli

doi:10.1155/2012/352493

Cited by 60 publications

(75 citation statements)

References 60 publications

(81 reference statements)

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“…The potential for OMV to serve as protective vaccines has been demonstrated for diverse bacteria (72)(73)(74)(75)(76). In addition, Pierson et al showed that vaccination with their OMV provided limited protection against subsequent F. novicida infection (32).…”

Section: Discussionmentioning

confidence: 99%

“…OMV have proved effective as vaccines for a number of bacterial pathogens (72)(73)(74)(75)(76), and Pierson et al showed that vaccination of mice with OMV isolated from late-stage F. novicida cultures provided limited protection against subsequent bacterial challenge (32). We vaccinated mice by intranasal administration of 20 g of purified OMV/T isolated from early-stationary-phase U112 cultures, or PBS as a control.…”

Section: Identification Of F Novicida Omv/t-associated Proteinsmentioning

confidence: 99%

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Production of Outer Membrane Vesicles and Outer Membrane Tubes by Francisella novicida

McCaig

Koller

Thanassi

2012

Journal of Bacteriology

128

148

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Francisella spp. are highly infectious and virulent bacteria that cause the zoonotic disease tularemia. Knowledge is lacking for the virulence factors expressed by Francisella and how these factors are secreted and delivered to host cells. Gram-negative bacteria constitutively release outer membrane vesicles (OMV), which may function in the delivery of virulence factors to host cells. We identified growth conditions under which Francisella novicida produces abundant OMV. Purification of the vesicles revealed the presence of tube-shaped vesicles in addition to typical spherical OMV, and examination of whole bacteria revealed the presence of tubes extending out from the bacterial surface. Recently, both prokaryotic and eukaryotic cells have been shown to produce membrane-enclosed projections, termed nanotubes, which appear to function in cell-cell communication and the exchange of molecules. In contrast to these previously characterized structures, the F. novicida tubes are produced in liquid as well as on solid medium and are derived from the OM rather than the cytoplasmic membrane. The production of the OMV and tubes (OMV/T) by F. novicida was coordinately regulated and responsive to both growth medium and growth phase. Proteomic analysis of purified OMV/T identified known Francisella virulence factors among the constituent proteins, suggesting roles for the vesicles in pathogenesis. In support of this, production of OM tubes by F. novicida was stimulated during infection of macrophages and addition of purified OMV/T to macrophages elicited increased release of proinflammatory cytokines. Finally, vaccination with purified OMV/T protected mice from subsequent challenge with highly lethal doses of F. novicida.

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Section: Discussionmentioning

confidence: 99%

Section: Identification Of F Novicida Omv/t-associated Proteinsmentioning

confidence: 99%

Production of Outer Membrane Vesicles and Outer Membrane Tubes by Francisella novicida

McCaig

Koller

Thanassi

2012

Journal of Bacteriology

128

148

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“…Considering the oral vaccine delivery, protection may be occurring through a highly localized response in the mucosa. A recent paper characterizing the protein content of Brucella melitensis outer membrane vesicles demonstrated the presence of a Dps protein in the vesicles (2). Since the protein is in outer membrane vesicles, this is probably the reason that protection against C. jejuni was partially generated following expression of the protein to the intestinal mucosa by the Salmonella vector.…”

Section: Discussionmentioning

confidence: 99%

“…To circumvent this problem, bacteria employ a variety of ferritins to meet their iron needs. In Campylobacter, this includes the genes dps, feoB, cfrA, ceuE, and Cj0178, which are involved with various aspects of iron acquisition (2,11,22,24,34).…”

mentioning

confidence: 99%

The Campylobacter jejuni Dps Homologue Is Important forIn VitroBiofilm Formation and Cecal Colonization of Poultry and May Serve as a Protective Antigen for Vaccination

Theoret

Cooper

Zekarias

et al. 2012

Clin Vaccine Immunol

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ABSTRACTIn this work, we investigated theCampylobacter jejuni dps(DNA binding protein from starved cells) gene for a role in biofilm formation and cecal colonization in poultry.In vitrobiofilm formation assays were conducted with stationary-phase cells in cell culture plates under microaerophilic conditions. These studies demonstrated a significant (>50%) reduction in biofilm formation by theC. jejuni dpsmutant compared to that by the wild-type strain. Studies in poultry also demonstrated the importance of thedpsgene in host colonization byC. jejuni. Real-time PCR analysis of mRNA extracted from the cecal contents of poultry infected with wild-typeC. jejuniindicated that thedpsgene is upregulated 20-fold during poultry colonization. Cecal colonization was greater than 5 log CFU lower in chicks infected with thedpsmutant than chicks infected with the wild-typeC. jejunistrain. Moreover, thedpsmutant failed to colonize 75% of the chicks following challenge with 105CFU. Preliminary studies were conducted in chicks by parenteral vaccination with a recombinant Dps protein or through oral vaccination with a recombinant attenuatedSalmonella entericastrain synthesizing theC. jejuniDps protein. No reduction inC. jejuniwas noted in chicks vaccinated with the parenteral recombinant protein, whereas, a 2.5-log-unit reduction ofC. jejuniwas achieved in chicks vaccinated with the attenuatedSalmonellavector after homologous challenge. Taken together, this work demonstrated the importance of Dps for biofilm formation and poultry colonization, and the study also provides a basis for continued work using the Dps protein as a vaccine antigen when delivered through aSalmonellavaccine vector.

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“…Recovery of crude exo-polysaccharide (exo-PS; fraction B1) followed the method published for B. melitensis (13), which sheds much of its outer membrane (14). Briefly, the previously noted supernatant was collected into a flask, and glacial acetic acid was added to 3% (vol/vol).…”

mentioning

confidence: 99%

Thermostable Cross-Protective Subunit Vaccine against Brucella Species

Cherwonogrodzky

Barabé

Grigat

et al. 2014

Clin Vaccine Immunol

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A subunit vaccine candidate was produced from Brucella suis 145 (biovar 4; expressing both the A antigen of Brucella abortus and the M antigen of Brucella melitensis). The preparation consisted mostly of polysaccharide (PS; >90% [wt/wt]; both cell-associated PS and exo-PS were combined) and a small amount of protein (1 to 3%) with no apparent nucleic acids. Vaccinated mice were protected (these had a statistically significant reduction in bacterial colonization compared to that of unvaccinated controls) when challenged with representative strains of three Brucella species most pathogenic for humans, i.e., B. abortus, B. melitensis, and B. suis. As little as 1 ng of the vaccine, without added adjuvant, protected mice against B. suis 145 infection (5 ؋ 10 5 CFU), and a single injection of 1 g of this subunit vaccine protected mice from B. suis 145 challenge for at least 14 months. A single immunization induced a serum IgG response to Brucella antigens that remained elevated for up to 9 weeks. The use of heat (i.e., boiling-water bath, autoclaving) in the vaccine preparation showed that it was thermostable. This method also ensured safety and security. The vaccine produced was immunogenic and highly protective against multiple strains of Brucella and represents a promising candidate for further evaluation.

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Characterization of Outer Membrane Vesicles fromBrucella melitensisand Protection Induced in Mice

Cited by 60 publications

References 60 publications

Production of Outer Membrane Vesicles and Outer Membrane Tubes by Francisella novicida

Production of Outer Membrane Vesicles and Outer Membrane Tubes by Francisella novicida

The Campylobacter jejuni Dps Homologue Is Important forIn VitroBiofilm Formation and Cecal Colonization of Poultry and May Serve as a Protective Antigen for Vaccination

Thermostable Cross-Protective Subunit Vaccine against Brucella Species

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