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Updated: Jul 9, 2025
Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
Published on: March 31, 2021
Characterization of outer membrane vesicles released by clinical isolates of Neisseria gonorrhoeae
Subhash Dhital1, Pankaj Deo1, Isabella Stuart1
1Department of Biochemistry & Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, Victoria, Australia.
Abstract:
The sexually transmitted pathogen Neisseria gonorrhoeae releases membrane vesicles including outer membrane vesicles (OMVs) during infections. OMVs traffic outer membrane molecules, such as the porin PorB and lipo-oligosaccharide (LOS), into host innate immune cells, eliciting 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 these vesicles similarly activate immune responses. Here, we characterized OMVs from four N. gonorrhoeae isolates and determined their size, abundance, proteome, LOS content, and activation of inflammatory responses in macrophages. The overall proteome of the OMVs was conserved between the four different isolates, which included major outer membrane and periplasm proteins. Despite this, we observed differences in the rate of OMV biogenesis and the relative abundance of 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-1α and IL-1β. Overall, these findings demonstrate that clinical isolates of N. gonorrhoeae utilize membrane vesicles to release proteins and lipids, which affects innate immune responses.
Insights
Neisseria gonorrhoeae releases outer membrane vesicles (OMVs) carrying molecules that trigger host cell death and inflammation. Differences in OMV composition from various infection sites cause varied immune responses.
Area of Science:
- Microbiology
- Immunology
- Cell Biology
Background:
- Neisseria gonorrhoeae, a sexually transmitted pathogen, releases outer membrane vesicles (OMVs).
- OMVs transport outer membrane molecules like porin PorB and lipo-oligosaccharide (LOS) into host cells.
- OMVs can induce host cell death and inflammation, but their composition and immune activation vary by clinical isolate.
Purpose of the Study:
- To characterize OMVs from different N. gonorrhoeae clinical isolates.
- To determine OMV proteome, LOS content, and their impact on immune responses.
Main Methods:
- Characterization of OMVs from four N. gonorrhoeae isolates.
- Analysis of OMV size, abundance, proteome, and LOS content.
- Assessment of inflammatory responses and cell death in macrophages.
Main Results:
- OMV proteome was conserved across isolates, including major outer membrane and periplasm proteins.
- Differences were observed in OMV biogenesis rates and relative abundance of membrane proteins and LOS.
- OMVs induced varying rates of macrophage cell death and secretion of IL-1α and IL-1β.
Conclusions:
- Clinical isolates of N. gonorrhoeae release OMVs with diverse molecular compositions.
- These OMVs differentially impact innate immune responses, including programmed cell death and cytokine secretion.
- Understanding OMV variations is crucial for gonorrhea pathogenesis and immune evasion strategies.
