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Detection and characterization of bacterial polysaccharides in drug-resistant enterococci

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Abstract

Enterococcus faecium (E. faecium) has emerged as one of today’s leading causes of health care-associated infections that is difficult to treat with the available antibiotics. These pathogens produce capsular polysaccharides on the cell surface which play a significant role in adhesion, virulence and evasion. Therefore, we aimed at the identification and characterization of bacterial polysaccharide antigens which are central for the development of vaccine-based prophylactic approaches. The crude cell wall-associated polysaccharides from E. faecium, its mutant and complemented strains were purified and analyzed by a primary antibody raised against lipoteichoic acid (LTA) and diheteroglycan (DHG). The resistant E. faecium strains presumably possess novel capsular polysaccharides that allow them to avoid the evasion from opsonic killing. The E. faecium U0317 strain was very well opsonized by anti-U0317 (~95%), an antibody against the whole bacterial cell. The deletion mutant showed a significantly increased susceptibility to opsonophagocytic killing (90–95%) against the penicillin binding protein (anti-PBP-5). By comparison, in a mouse urinary tract and rat endocarditis infection model, respectively, there were no significant differences in virulence. In this study we explored the biological role of the capsule of E. faecium. Our findings showed that the U0317 strain is not only sensitive to anti-LTA but also to antibodies against other enterococcal surface proteins. Our findings demonstrate that polysaccharides capsule mediated-resistance to opsonophagocytosis. We also found that the capsular polysaccharides do not play an important role in bacterial virulence in urinary tract and infective endocarditis in vivo models.

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Abbreviations

CPS :

Capsular polysaccharides

CFU :

Colony-forming unit

DHG :

Diheteroglycan

E. faecium :

Enterococcus faecium

LTA :

Lipoteichoic Acid

OPA :

Opsonophagocytic Assays

PBP :

Penicillin Binding Protein-5

UTI :

Urinary Tract Infection

VRE :

Vancomycin-Resistant Enterococci

Wt :

Wild-type

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Acknowledgments

LA sincerely thanks the Deutscher Akademischer AustauschDienst (DAAD) for the award of a PhD fellowship and Dominique Wobser for help with the animal experiments.

Funding

This work was supported by grants from the German Ministry of Science and Education (BMBF: UroGenOmics0315833C).

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Authors and Affiliations

  1. Division of Infectious Diseases, Department of Internal Medicine, University Medical Center Freiburg, Hugstetter Straße 55, D-79106, Freiburg, Germany

    Liaqat Ali, Hubert E. Blum & Türkân Sakιnç

  2. Department of Biological Sciences, National University of Medical Sciences (NUMS), Rawalpindi, 46000, Pakistan

    Liaqat Ali

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  1. Liaqat Ali
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  2. Hubert E. Blum
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  3. Türkân Sakιnç
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LA and TS conceived and designed the study. LA analyzed the data and drafted the manuscript. HEB critically reviewed the manuscript. All the authors studied and approved the final manuscript.

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Correspondence to Liaqat Ali.

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The authors declare that they have no competing interests.

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All animal experiments were performed in compliance with the German animal protection law (TierSchG). The mice were housed and handled following good animal practice as defined by FELASA and the national animal welfare body GV-SOLAS. The animal welfare committees of the University of Freiburg (Regierungspräsidium Freiburg Az 35/9185.81/G-11/118 and Az 35/9185.81/G-12/070) approved all animal experiments.

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Ali, L., Blum, H.E. & Sakιnç, T. Detection and characterization of bacterial polysaccharides in drug-resistant enterococci. Glycoconj J 36, 429–438 (2019). https://doi.org/10.1007/s10719-019-09881-3

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