Comparative proteomic analysis of outer membrane vesicles from Brucella suis, Brucella ovis, Brucella canis and Brucella neotomae

Ruiz-Palma, María Del Socorro; Ávila-Calderón, Eric Daniel; Aguilera‐Arreola, Ma. Guadalupe; López-Merino, Ahidé; Ruíz, Enrico A.; Morales-García, María del Rosario J.; López-Villegas, Edgar Óliver; Gómez-Lunar, Zulema; Arellano-Reynoso, Beatriz; Contreras-Rodríguez, Araceli

doi:10.1007/s00203-020-02170-w

Cited by 12 publications

(9 citation statements)

References 74 publications

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“…Other interesting structures promoting adhesiveness to solid surfaces in Brucella are the OM vesicles (OMVs), commonly called blebs (Godefroid et al, 2010). Blebs are nanovesicles released from the OM whose composition (Gamazo and Moriyón, 1987;Avila-Calderón et al, 2020;Ruiz-Palma et al, 2021) and amount (Solanki et al, 2021) have been associated with the lack of O-PS, and they are overproduced in strains that auto-agglutinate (Godefroid et al, 2010). As shown by TEM (Figure 3G), the mutants showed abundant OMVs as outward bulges protruding from the surface or as already detached vesicles, while the OM maintained its integrity.…”

Section: Blocking O-polysaccharide Transport Causes Pleiotropic Chang...mentioning

confidence: 99%

Brucella melitensis Wzm/Wzt System: Changes in the Bacterial Envelope Lead to Improved Rev1Δwzm Vaccine Properties

et al. 2022

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The lipopolysaccharide (LPS) O-polysaccharide (O-PS) is the main virulence factor in Brucella. After synthesis in the cytoplasmic membrane, O-PS is exported to the periplasm by the Wzm/Wzt system, where it is assembled into a LPS. This translocation also engages a bactoprenol carrier required for further biosynthesis pathways, such as cell wall biogenesis. Targeting O-PS export by blockage holds great potential for vaccine development, but little is known about the biological implications of each Wzm/Wzt moiety. To improve this knowledge and to elucidate its potential application as a vaccine, we constructed and studied wzm/wzt single- and double-deletion mutants, using the attenuated strain Brucella melitensis Rev1 as the parental strain. This allowed us to describe the composition of Brucella peptidoglycan for the first time. We observed that these mutants lack external O-PS yet trigger changes in genetic transcription and in phenotypic properties associated with the outer membrane and cell wall. Both mutants are highly attenuated; unexpectedly, Rev1Δwzm also excels as an immunogenic and effective vaccine against B. melitensis and Brucella ovis in mice, revealing that low persistence is not at odds with efficacy. Rev1Δwzm is attenuated in BeWo trophoblasts, does not infect mouse placentas, and is safe in pregnant ewes. Overall, these attributes and the minimal serological interference induced in sheep make Rev1Δwzm a highly promising vaccine candidate.

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Section: Blocking O-polysaccharide Transport Causes Pleiotropic Chang...mentioning

confidence: 99%

Brucella melitensis Wzm/Wzt System: Changes in the Bacterial Envelope Lead to Improved Rev1Δwzm Vaccine Properties

et al. 2022

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“…BamD deletion causes lethality in E. coli and Neisseria meningitidis , and Bam has a role in the production of OMPs for survival and pathogenesis [59]. Proteins Omp25, Omp31 and SodC were also identified: these proteins have been well characterized as virulence factors or immunogenic proteins in Brucella ; further these proteins were identified in outer membrane vesicles (OMVs) in B. canis [60]. The protein Sod (Superoxide dismutase [Cu‐Zn]) is associated to virulence in a number of microorganisms [31].…”

Section: Discussionmentioning

confidence: 99%

Proteomics and bioinformatics investigations to improve serological diagnosis of canine brucellosis

Luciani

Krasteva

Febo

et al. 2023

Proteomics Clinical Apps

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PurposeBrucella canis is pathogenic for dogs and humans. Serological diagnosis is a cost‐effective approach for disease surveillance, but a major drawback of current serological tests is the cross‐reactivity with other bacteria that results in false positive reactions. Development of indirect tests with improved sensitivity and specificity that use selected B. canis proteins instead of the whole antigen remain a priority.Experimental DesignA western blotting assay was developed to define the serum antibody patterns associated to infection using a panel of positive and negative dog sera. B. canis positive sera recognized immunogenic bands ranging from 7 to 30 kDa that were then submitted to ESI–LC‐MS/MS and analyzed by bioinformatics tools.ResultsA total of 398 B. canis proteins were identified. Bioinformatics tools identified 16 non cytoplasmic immunogenic proteins predicted as non‐homologous with the most important Brucella cross‐reactive bacteria and nine B. canis proteins non‐homologous to B. ovis; among the latter, one resulted non‐homologous to B. melitensis. Data are available via ProteomeXchange with identifier PXD042682.Conclusions and Clinical RelevanceThe western blotting test developed was able to distinguish between infected and non‐infected animals and may serve as a confirmatory test for the serological diagnosis of B. canis. The mass spectrometry and in silico results lead to the identification of specific candidate antigens that pave the way for the development of more accurate indirect diagnostic tests.

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“…S. epidermidis clinical strain 983 was isolated from a corneal infection; it is an ST2 genotype and biofilm producer [ 68 ]. EVs from 983 and ATCC12228 strains were obtained according to Avila-Calderón et al (2012) and Ruiz-Palma et al (2021) under the same culture conditions [ 69 , 70 ]. Briefly, both strains were cultured on supplemented LB agar plates and incubated for 24 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

The Extracellular Vesicles from the Commensal Staphylococcus Epidermidis ATCC12228 Strain Regulate Skin Inflammation in the Imiquimod-Induced Psoriasis Murine Model

Gómez‐Chávez

Cedillo-Peláez

Zapi‑Colín

et al. 2021

IJMS

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Extracellular vesicles (EVs) are evaginations of the cytoplasmic membrane, containing nucleic acids, proteins, lipids, enzymes, and toxins. EVs participate in various bacterial physiological processes. Staphylococcus epidermidis interacts and communicates with the host skin. S. epidermidis’ EVs may have an essential role in this communication mechanism, modulating the immunological environment. This work aimed to evaluate if S. epidermidis’ EVs can modulate cytokine production by keratinocytes in vitro and in vivo using the imiquimod-induced psoriasis murine model. S. epidermidis’ EVs were obtained from a commensal strain (ATC12228EVs) and a clinical isolated strain (983EVs). EVs from both origins induced IL-6 expression in HaCaT keratinocyte cultures; nevertheless, 983EVs promoted a higher expression of the pro-inflammatory cytokines VEGF-A, LL37, IL-8, and IL-17F than ATCC12228EVs. Moreover, in vivo imiquimod-induced psoriatic skin treated with ATCC12228EVs reduced the characteristic psoriatic skin features, such as acanthosis and cellular infiltrate, as well as VEGF-A, IL-6, KC, IL-23, IL-17F, IL-36γ, and IL-36R expression in a more efficient manner than 983EVs; however, in contrast, Foxp3 expression did not significantly change, and IL-36 receptor antagonist (IL-36Ra) was found to be increased. Our findings showed a distinctive immunological profile induction that is dependent on the clinical or commensal EV origin in a mice model of skin-like psoriasis. Characteristically, proteomics analysis showed differences in the EVs protein content, dependent on origin of the isolated EVs. Specifically, in ATCC12228EVs, we found the proteins glutamate dehydrogenase, ornithine carbamoyltransferase, arginine deiminase, carbamate kinase, catalase, superoxide dismutase, phenol-soluble β1/β2 modulin, and polyglycerol phosphate α-glucosyltransferase, which could be involved in the reduction of lesions in the murine imiquimod-induced psoriasis skin. Our results show that the commensal ATCC12228EVs have a greater protective/attenuating effect on the murine imiquimod-induced psoriasis by inducing IL-36Ra expression in comparison with EVs from a clinical isolate of S. epidermidis.

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Comparative proteomic analysis of outer membrane vesicles from Brucella suis, Brucella ovis, Brucella canis and Brucella neotomae

Cited by 12 publications

References 74 publications

Brucella melitensis Wzm/Wzt System: Changes in the Bacterial Envelope Lead to Improved Rev1Δwzm Vaccine Properties

Brucella melitensis Wzm/Wzt System: Changes in the Bacterial Envelope Lead to Improved Rev1Δwzm Vaccine Properties

Proteomics and bioinformatics investigations to improve serological diagnosis of canine brucellosis

The Extracellular Vesicles from the Commensal Staphylococcus Epidermidis ATCC12228 Strain Regulate Skin Inflammation in the Imiquimod-Induced Psoriasis Murine Model

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