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Characterization of outer membrane vesicles released by clinical isolates of Neisseria gonorrhoeae

ABSTRACT: The sexually transmitted pathogen Neisseria gonorrhoeae releases outer membrane vesicles (OMVs) during infections. OMVs traffic the major porin PorB, other membrane proteins and lipo-oligosaccharide (LOS) into host innate immune cells and activate programmed cell death pathways and inflammation. Little is known, however, about the proteome and LOS content of OMVs released by clinical strains isolated from different infection sites, and whether this affects immune responses. Here, we characterized OMVs from four N. gonorrhoeae isolates and determined their size, abundance, proteome and activation of inflammatory responses in macrophages. The overall proteome of the OMVs was conserved between the four different isolates, included major outer membrane, periplasm, cytoplasmic membrane proteins. Despite this, we observed differences in the rate of OMV biogenesis and the relative abundance of major outer membrane proteins and LOS. Consequently, OMVs from clinical isolates induced varying rates of macrophage cell death and the secretion of interleukin-1 family members, such as Il-1andIl-1. Overall, these findings demonstrate that clinical isolates of N. gonorrhoeae utilize OMVs to release major proteins and lipids, which affects innate immune responses.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Neisseria Gonorrhoeae

TISSUE(S): Cell Culture

SUBMITTER: Ralf Schittenhelm

LAB HEAD: Thomas Naderer

PROVIDER: PXD042787 | Pride | 2024-08-27

REPOSITORIES: Pride

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Project description:Tenacibaculum maritimum, the etiological agent of tenacibaculosis in marine fish, constitutively secretes extracellular products (ECPs), which play a main role in virulence. However, their protein content has not been yet comprehensively studied. In this study a collection of 64 T. maritimum strains belonging to the four serotypes described so far (O1 to O4) was used to analyse the prevalence of extracellular proteolytic and lipolytic activities related to virulence. Results showed the existence of a large heterogeneity of the enzymatic capacity among serotype O4 isolates. Most notably, the ECPs of T. maritimum SP9.1 belonging to serotype O4 contain a large amount of outer membrane vesicles (OMVs), which were characterized by electron microscopy and purified by successive steps of filtration and centrifugation. Total protein content of ECPs, and the proteins associated to OMVs and soluble fraction of the ECPs (S-ECPs) were identified by nLC-TIMS-QTOF. A total of 641 proteins were identified in ECPs including some virulence related factors, which were mainly found in one of the fractions, either in OMVs or S-ECPs. Outer membrane proteins such as TonB-dependent transporters and the T9SS-related proteins PorP, PorT and SprA appeared to be mainly associated with OMVs. By contrast, putative virulence factors such as sialidase SiaA, chondroitinase CslA, sphingomyelinase Sph, ceramidase Cer and collagenase Col were found only in the S-ECPs. These findings demonstrate that T. maritimum release, through surface blebbing, outer membrane vesicles that are enriched specifically in TonB-dependent siderophore transporters and proteins of the type IX secretion system. Interestingly, our results also showed that OMVs could play a role in virulence by promoting surface adhesion, biofilm formation, and maximizing cytotoxic effects of the ECPs. The T. maritimum secretome analysis provides insights into the extracellular products function and can constitute the basis for future studies aimed to elucidate the role of OMVs in the pathogenesis of fish tenacibaculosis.

			Action	DRS
	P1CH20190308_DDA_14A.raw	Raw
	P1CH20190308_DDA_14B.raw	Raw
	P1CH20190308_DDA_17A.raw	Raw
	P1CH20190308_DDA_17B.raw	Raw
	P1CH20190308_DDA_20A.raw	Raw

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Characterization of outer membrane vesicles released by clinical isolates of Neisseria gonorrhoeae

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