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Environmental influences on Streptococcus sanguinis membrane vesicle biogenesis

The ISME Journal volume 17, pages 1430–1444 (2023)Cite this article

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Abstract

Membrane vesicles are produced by Gram-negative and Gram-positive bacteria. While membrane vesicles are potent elicitors of eukaryotic cells and involved in cell-cell communication, information is scarce about their general biology in the context of community members and the environment. Streptococcus sanguinis, a Gram-positive oral commensal, is prevalent in the oral cavity and well-characterized for its ability to antagonize oral pathobionts. We have found that production and dissemination of membrane vesicles by S. sanguinis is dependent on environmental and community factors. Co-culture with interacting commensal Corynebacterium durum, as well as with the periodontal pathobiont Filifactor alocis had no effect on S. sanguinis vesicle number and size, whereas the periodontal pathobiont Porphyromonas gingivalis abolished S. sanguinis vesicle production. Using both correlation and differential expression analyses to examine the transcriptomic changes underlying vesicle production, we found that differential expression of genes encoding proteins related to the cytoplasmic membrane and peptidoglycan correlate with the abundance of membrane vesicles. Proteomic characterizations of the vesicle cargo identified a variety of proteins, including those predicted to influence host interactions or host immune responses. Cell culture studies of gingival epithelial cells demonstrated that both crude and highly purified membrane vesicles could induce the expression of IL-8, TNF-α, IL-1β, and Gro-α within 6 hours of inoculation at levels comparable to whole cells. Our findings suggest that production of membrane vesicles by S. sanguinis is heavily influenced by community and environmental factors and plays an important role in communication with host cells.

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Fig. 1: S. sanguinis produces membrane vesicles in a dual culture with C. durum in media supplemented with glucose.
Fig. 2: S. sanguinis vesicle production is varied in the presence of pathobionts F. alocis or P. gingivalis.
Fig. 3: Transcriptomic changes associated with MV number.
Fig. 4: Comparison of differential expression and correlation analysis.
Fig. 5: Proteomic characterization of S. sanguinis MVs.
Fig. 6: Functional associations of proteins predicted in S. sanguinis MV cargo.
Fig. 7: Inoculation of gingival epithelial cells with S. sanguinis membrane vesicles induced expression of cytokines.
Fig. 8: Gingival epithelial cell visualization and MV internalization.

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Data availability

All raw and normalized RNA sequence data and associated metadata are available on NCBI Gene Expression Omnibus (GEO), under accession GSE225861. The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [1] partner repository with the dataset identifier PXD041791.

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Acknowledgements

We would like to thank the following: OHSU Massive Parallel Sequencing Core (Robert Searles, Amy Carlos), OHSU Microscropy Facility (Claudia Lopez, Steven Adamou), OHSU Imaging Facility (Stefanie Kaech Petrie) and Oregon State University Proteomics Core Facility (Stanislau Stanisheuski). We are grateful to Özlem Yilmaz (Medical University of South Carolina) for sharing F. alocis, Richard J. Lamont (University of Louisville) for sharing P. gingivalis gingipain mutants, and Delaney Shea for assistance with nanoparticle tracking analysis. This work was supported by an NIH-NIDCR grant DE021726, DE029492 and DE029612 to JK and NIH-NIDCR grant DE028252 to J.M.

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Authors and Affiliations

  1. Department of Restorative Dentistry, School of Dentistry, Oregon Health & Science University (OHSU), Portland, OR, USA

    Emily Helliwell, Justin Merritt & Jens Kreth

  2. Department of Community Dentistry, School of Dentistry, Oregon Health & Science University (OHSU), Portland, OR, USA

    Dongseok Choi

  3. School of Public Health, Oregon Health & Science University (OHSU), Portland, OR, USA

    Dongseok Choi

  4. Department of Molecular Microbiology and Immunology, Oregon Health & Science University (OHSU), Portland, OR, USA

    Justin Merritt & Jens Kreth

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E.H., J.M., and J.K .developed and designed the research. EH performed the experiments. E.H., D.C., and JK analyzed data. E.H. drafted the manuscript. E.H., D.C., J.M. and J.K. edited and revised the manuscript. All authors reviewed and approved the results and the revisions made to the manuscript.

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Helliwell, E., Choi, D., Merritt, J. et al. Environmental influences on Streptococcus sanguinis membrane vesicle biogenesis. ISME J 17, 1430–1444 (2023). https://doi.org/10.1038/s41396-023-01456-3

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