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An antibacterial vaccination strategy based on a glycoconjugate containing the core lipopolysaccharide tetrasaccharide Hep2Kdo2

Nature Chemistry volume 8, pages 242–249 (2016)Cite this article

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Abstract

Certain non-mammalian cell wall sugars are conserved across a variety of pathogenic bacteria. This conservation of structure, combined with their structural differences when compared with mammalian sugars, make them potentially powerful epitopes for immunization. Here, we report the synthesis of a glycoconjugate that displays the so-called ‘inner core’ sugars of Gram-negative bacterial cell walls. We also describe an antibacterial vaccination strategy based on immunization with the glycoconjugate and the subsequent administration of an inhibitor that uncovers the corresponding epitope in pathogenic bacteria. The core tetrasaccharide, Hep2Kdo2, a common motif in bacterial lipopolysaccharides, was synthesized and attached via a chain linker to a diphtheria toxin mutant carrier protein. This glycoconjugate generated titres of antibodies towards the inner core tetrasaccharide of the lipopolysaccharide, which were capable of binding the cell-surface sugars of bacterial pathogenic strains including Neisseria meningitidis, Pseudomonas aeruginosa and Escherichia coli. Exposure of bacterial lipopolysaccharide in in vitro experiments, using an inhibitor of capsular polysaccharide transport, enabled potent bacterial killing with antiserum.

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Figure 1: The target core tetrasaccharide of the lipooligosaccharide (LOS) of Gram-negative bacteria.
Figure 2: Design of glycoconjugate with the core tetrasaccharide of the LOS of Gram-negative bacteria.
Figure 3: Synthesis of glycoconjugate vaccine 1.
Figure 4: Mass spectrometric (MALDI-TOF) analysis of glycoconjugate 1.
Figure 5: Immune responses and antibacterial activity from the putative vaccine glycoconjugate 1.

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Acknowledgements

The authors thank R. Barrow and X. Bai (Public Health England) for performing SBA assays, G. Lipowsky for managing immunization experiments and T. Claridge for assistance with NMR analyses. The authors acknowledge GlycoVaxyn for financial support. B.G.D. was a Royal Society Wolfson Research Merit Award recipient.

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Author notes

  1. Michael Kowarik, Larissa Neiwert & Amirreza Faridmoayer

    Present address: Present address: LimmaTech Biologics AG, Grabenstrasse 3, Schlieren 8952, Switzerland,

  2. Lingbing Kong and Balakumar Vijayakrishnan: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford, OX1 3TA, UK

    Lingbing Kong, Balakumar Vijayakrishnan, Jin Park, Alexandra N. Zakharova & Benjamin G. Davis

  2. GlycoVaxyn AG, Grabenstrasse 3, Schlieren, 8952, Switzerland

    Michael Kowarik, Larissa Neiwert & Amirreza Faridmoayer

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Contributions

L.K., B.V., A.F., M.K. and B.G.D. designed the experiments. L.K., J.P., A.N.Z. and B.V. conducted the carbohydrate syntheses. B.V. generated protein scaffolds and constructed the corresponding glycoconjugates. L.K., B.V. and L.N. performed the immunological experiments. L.K., B.V., M.K. and B.G.D. analysed the results. L.K., B.V. and B.G.D. wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to Benjamin G. Davis.

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Competing interests

M.K., L.N. and A.F. are employed by GlycoVaxyn.

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Kong, L., Vijayakrishnan, B., Kowarik, M. et al. An antibacterial vaccination strategy based on a glycoconjugate containing the core lipopolysaccharide tetrasaccharide Hep2Kdo2. Nature Chem 8, 242–249 (2016). https://doi.org/10.1038/nchem.2432

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