High Yield Production Process for Shigella Outer Membrane Particles

Scorza, Francesco Berlanda; Colucci, Anna; Maggiore, Luana; Sanzone, Silvia; Rossi, Omar; Ferlenghi, Ilaria; Pesce, Isabella; Caboni, Mariaelena; Norais, Nathalie; Cioccio, Vito Di; Saul, Allan; Gerke, Christiane

doi:10.1371/journal.pone.0035616

Cited by 124 publications

(158 citation statements)

References 43 publications

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“…This was somewhat surprising, considering that cytoplasmic and IM proteins are generally not present or are present only at very low levels in OMVs (4,56). However, GroEL and members of the PDH complex have frequently been detected by proteomics in OMV preparations (57)(58)(59)(60). Our analyses showed that GroEL and PDH-E1 were loosely associated with the OMV surface.…”

Section: Discussionmentioning

confidence: 71%

“…Furthermore, OMV-associated GroEL was detected as an ϳ700-kDa complex upon blue native PAGE (data not shown), consistent with its cytoplasmic tetradecameric form (61). Therefore, and given their frequent association with OMVs and bacterial surfaces (57)(58)(59)(60)62), it is likely that GroEL and PDH-E1 leak from cells under stress conditions and reassociate with bacterial outer membranes or vesicles. Indeed, mild stress could be triggered by the overexpression of Hbp, a notion that is supported by the concomitant upregulation of the outer membrane lipoprotein OsmB, which is encoded by a multistress-responsive gene (63), and of OmpX, which is also known to respond to stress conditions (64).…”

Section: Discussionmentioning

confidence: 76%

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Decoration of Outer Membrane Vesicles with Multiple Antigens by Using an Autotransporter Approach

Daleke-Schermerhorn¹,

Félix

Soprova³

et al. 2014

Appl Environ Microbiol

101

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iOuter membrane vesicles (OMVs) are spherical nanoparticles that naturally shed from Gram-negative bacteria. They are rich in immunostimulatory proteins and lipopolysaccharide but do not replicate, which increases their safety profile and renders them attractive vaccine vectors. By packaging foreign polypeptides in OMVs, specific immune responses can be raised toward heterologous antigens in the context of an intrinsic adjuvant. Antigens exposed at the vesicle surface have been suggested to elicit protection superior to that from antigens concealed inside OMVs, but hitherto robust methods for targeting heterologous proteins to the OMV surface have been lacking. We have exploited our previously developed hemoglobin protease (Hbp) autotransporter platform for display of heterologous polypeptides at the OMV surface. One, two, or three of the Mycobacterium tuberculosis antigens ESAT6, Ag85B, and Rv2660c were targeted to the surface of Escherichia coli OMVs upon fusion to Hbp. Furthermore, a hypervesiculating ⌬tolR ⌬tolA derivative of attenuated Salmonella enterica serovar Typhimurium SL3261 was generated, enabling efficient release and purification of OMVs decorated with multiple heterologous antigens, exemplified by the M. tuberculosis antigens and epitopes from Chlamydia trachomatis major outer membrane protein (MOMP). Also, we showed that delivery of Salmonella OMVs displaying Ag85B to antigen-presenting cells in vitro results in processing and presentation of an epitope that is functionally recognized by Ag85B-specific T cell hybridomas. In conclusion, the Hbp platform mediates efficient display of (multiple) heterologous antigens, individually or combined within one molecule, at the surface of OMVs. Detection of antigen-specific immune responses upon vesicle-mediated delivery demonstrated the potential of our system for vaccine development.

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Section: Discussionmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 76%

Decoration of Outer Membrane Vesicles with Multiple Antigens by Using an Autotransporter Approach

Daleke-Schermerhorn¹,

Félix

Soprova³

et al. 2014

Appl Environ Microbiol

101

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“…Two subunit parenteral vaccines are in development to prevent invasive NTS infections among young children in sub-Saharan Africa, including a conjugate consisting of core plus O polysaccharide linked to phase 1 flagellin subunits of the homologous serovar and another consisting of outer membrane particles, called Generalized Module for Membrane Antigens (GMMA) (Gendrivax [http://www.gendrivax.org]) (31,33). Administered parenterally, these vaccines are expected to protect by eliciting functional (likely IgG) antibodies that intercept the invading bacteria during primary bacteremia.…”

Section: Discussionmentioning

confidence: 99%

Refined Live Attenuated Salmonella enterica Serovar Typhimurium and Enteritidis Vaccines Mediate Homologous and Heterologous Serogroup Protection in Mice

et al. 2015

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Invasive nontyphoidal Salmonella (NTS) infections constitute a major health problem among infants and toddlers in sub-Saharan Africa; these infections also occur in infants and the elderly in developed countries. We genetically engineered a Salmonella enterica serovar Typhimurium strain of multilocus sequence type 313, the predominant genotype circulating in sub-Saharan Africa. We evaluated the capacities of S. Typhimurium and Salmonella enterica serovar Enteritidis ⌬guaBA ⌬clpX live oral vaccines to protect mice against a highly lethal challenge dose of the homologous serovar and determined protection against other group B and D serovars circulating in sub-Saharan Africa. The vaccines S. Typhimurium CVD 1931 and S. Enteritidis CVD 1944 were immunogenic and protected BALB/c mice against 10,000 50% lethal doses (LD 50 ) of S. Typhimurium or S. Enteritidis, respectively. S. Typhimurium CVD 1931 protected mice against the group B serovar Salmonella enterica serovar Stanleyville (91% vaccine efficacy), and S. Enteritidis CVD 1944 protected mice against the group D serovar Salmonella enterica serovar Dublin (85% vaccine efficacy). High rates of survival were observed when mice were infected 12 weeks postimmunization, indicating that the vaccines elicited long-lived protective immunity. Whereas CVD 1931 did not protect against S. Enteritidis R11, CVD 1944 did mediate protection against S. Typhimurium D65 (81% efficacy). These findings suggest that a bivalent (S. Typhimurium and S. Enteritidis) vaccine would provide broad protection against the majority of invasive NTS infections in sub-Saharan Africa. N on-typhoidal Salmonella (NTS) is a leading cause of bacterial bloodstream infections in febrile children and immunocompromised individuals in sub-Saharan Africa and has been associated with a high case fatality rate of 20 to 25% (1). Although severe malarial anemia and human immunodeficiency virus (HIV) are important risk factors for invasive NTS infection, the disease is also common in low-HIV-prevalence areas (1-4).There are Ͼ2,500 Salmonella serovars that can be differentiated on the basis of the O polysaccharide (OPS) antigens of their lipopolysaccharide (LPS) and their H flagellum antigens, using the Kauffman-White typing scheme (5). For example, Salmonella enterica serovar Typhimurium has O antigens 1, 4, 12, and occasionally 5. Epitope 12 is formed by trisaccharide repeats of mannose, rhamnose, and galactose; glucosylation of the galactose residue forms epitope 1. An abequose linked to each mannose defines it as a serovar within group B and constitutes the immunodominant O4 epitope; epitope 5 results from a phage conversion that introduces an O-acetyl moiety on the abequose. S. Typhimurium also has separate H antigens, i (phase 1) and 1 and 2 (phase 2), expressed alternatively via a process called phase variation. In contrast, Salmonella enterica serovar Enteritidis has O antigen epitope 9, which identifies it as a member of group D. Epitope 9 is formed by a tyvelose residue that is linked to the mannose of the ...

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“…2011,2013,2014). OMVs may also contain toxins and other virulence factors (Kulp & Kuehn, 2010 Studies on outer membrane vesicles after imposition of different stresses were found to be different (Berlanda Scorza et al, 2012). OMVs have been used successfully as delivery vehicles for recombinant proteins (Chen et al, 2010).…”

Section: Proteinsmentioning

confidence: 99%

Biogenesis and multifaceted roles of outer membrane vesicles from Gram-negative bacteria

2014

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Outer membrane vesicles (OMVs) released from Gram-negative bacteria consist of lipids, proteins, lipopolysaccharides and other molecules. OMVs are associated with several biological functions such as horizontal gene transfer, intracellular and intercellular communication, transfer of contents to host cells, and eliciting an immune response in host cells. Although hypotheses have been made concerning the mechanism of biogenesis of these vesicles, research on OMV formation is far from complete. The roles of outer membrane components, bacterial quorum sensing molecules and some specific proteins in OMV biogenesis have been studied. This review discusses the different models that have been proposed for OMV biogenesis, along with details of the biological functions of OMVs and the likely scope of future research.

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High Yield Production Process for Shigella Outer Membrane Particles

Cited by 124 publications

References 43 publications

Decoration of Outer Membrane Vesicles with Multiple Antigens by Using an Autotransporter Approach

Decoration of Outer Membrane Vesicles with Multiple Antigens by Using an Autotransporter Approach

Refined Live Attenuated Salmonella enterica Serovar Typhimurium and Enteritidis Vaccines Mediate Homologous and Heterologous Serogroup Protection in Mice

Biogenesis and multifaceted roles of outer membrane vesicles from Gram-negative bacteria

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