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      Structural characterisation of the capsular polysaccharide expressed by Burkholderia thailandensis strain E555:: wbiI (pKnock-KmR) and assessment of the significance of the 2-O-acetyl group in immune protection

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      Date
      2017-09-21
      Author
      Bayliss, M
      Donaldson, MI
      Nepogodiev, SA
      Pergolizzi, G
      Scott, AE
      Harmer, NJ
      Field, RA
      Prior, JL
      Date issued
      2017-09-21
      Journal
      Carbohydrate Research
      Type
      Article
      Language
      en
      Publisher
      Elsevier
      Links
      https://www.ncbi.nlm.nih.gov/pubmed/29024844
      Rights
      Open Access funded by Biotechnology and Biological Sciences Research Council under a Creative Commons license: https://creativecommons.org/licenses/by/4.0/. Content includes material subject to © Crown copyright (2017), Dstl. This material is licensed under the terms of the Open Government Licence except where otherwise stated. To view this licence, visit http://www.nationalarchives.gov.uk/doc/opengovernment-licence/version/3 or write to the Information Policy Team, The National Archives, Kew, London TW9 4DU, or email: psi@nationalarchives.gsi.gov.uk.
      Abstract
      Burkholderia pseudomallei and its close relative B. mallei are human pathogens that are classified as Tier 1 bio-threat agents. Both organisms have previously been shown to constitutively produce a capsular polysaccharide (CPS) that is both a virulence determinant and protective antigen. Extraction and purification of CPS for use as a potential vaccine candidate requires containment level 3 laboratories which is expensive and time-consuming. B. thailandensis strain E555 is closely related to B. pseudomallei and B. mallei, but is non-pathogenic to humans and based on immunological cross-reactivity has previously been shown to express a B. pseudomallei-like CPS. In this study, capsular polysaccharide isolated from an O-antigen deficient strain of B. thailandensis E555 was identified by 1H and 13C NMR spectroscopy as -3-)-2-O-acetyl-6-deoxy-β-d-manno-heptopyranose-(-1, and identical to that produced by B. pseudomallei. This was further substantiated by anti-CPS monoclonal antibody binding. In connection with the production of CPS fragments for use in glycoconjugate vaccines, we set out to assess the importance or otherwise of the CPS 2-OAc groups in immune protection. To this end conjugates of the native and de-O-acetylated CPS with the Hc fragment of tetanus toxin (TetHc) were used as vaccines in a mouse model of melioidosis. The level of protection provided by deacetylated CPS was significantly lower than that from native, acetylated CPS. In addition, sera from mice vaccinated with the deacetylated CPS conjugate did not recognise native CPS. This suggests that CPS extracted from B. thailandensis can be used as antigen and that the acetyl group is essential for protection.
      Funders/Sponsor
      This research was funded by the US Defence Threat Reduction Agency (DTRA), grant CBBAA12-VAXBT2-1-0032 and the United Kingdom Ministry of Defence. Studies at the JIC were supported by the UK BBSRC Institute Strategic Programme on Understanding and Exploiting Metabolism (MET) [BB/J004561/1] and the John Innes Foundation. MID was supported by a BBSRC Industrial CASE award with Mologic Ltd.
      Description
      This is the author accepted manuscript. The final version is available from Elsevier via the DOI in this record.
      Citation
      Vol. 452, pp. 17 - 24
      DOI
      https://doi.org/10.1016/j.carres.2017.09.011
      URI
      http://hdl.handle.net/10871/30662
      Collections
      • Biosciences
      Place of publication
      Netherlands

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