Lipopolysaccharide associated with β-2,6 fructan mediates TLR4-dependent immunomodulatory activity in vitro

Young, Ian D., Nepogodiev, Sergey A., Black, Ian M., Le Gall, Gwenaelle, Wittmann, Alexandra, Latousakis, Dimitrios, Visnapuu, Triinu, Azadi, Parastoo, Field, Robert A., Juge, Nathalie and Kawasaki, Norihito (2021) Lipopolysaccharide associated with β-2,6 fructan mediates TLR4-dependent immunomodulatory activity in vitro. Carbohydrate Polymers. ISSN 0144-8617

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Abstract

Levan, a β-2,6 fructofuranose polymer produced by microbial species, has been reported for its immunomodulatory properties via interaction with toll-like receptor 4 (TLR4) which recognises lipopolysaccharide (LPS). However, the molecular mechanisms underlying these interactions remain elusive. Here, we investigated the immunomodulatory properties of levan using thoroughly-purified and characterised samples from Erwinia herbicola and other sources. E. herbicola levan was purified by gel-permeation chromatography and LPS was removed from the levan following a novel alkali treatment developed in this study. E. herbicola levan was then characterised by gas chromatography–mass spectrometry and NMR. We found that levan containing LPS, but not LPS-depleted levan, induced TLR4-mediated cytokine production by bone marrow-derived dendritic cells and/or activated TLR4 reporter cells. These data indicated that the immunomodulatory properties of the levan toward TLR4-expressing immune cells were mediated by the LPS. This work also demonstrates the importance of LPS removal when assessing the immunomodulatory activity of polysaccharides.

Item Type: Article
Additional Information: Funding Information: The authors gratefully acknowledge the support of the Biotechnology and Biological Sciences Research Council (BBSRC); this research was funded by the BBSRC Institute Strategic Programme Grant Food and Health BBS/E/F/00044486, Food Innovation and Health BB/R012512/1, Gut Microbes and Health BB/R012490/1 and its constituent project(s), and Molecules from Nature - Products and Pathways BBS/E/J/000PR9790. Ian D. Young held a BBSRC Doctoral Training Partnership studentship (Project ID: BB/JO14524/1). Norihito Kawasaki would like to thank the Marie-Curie International Incoming Fellowship from the European Union 7th Framework Programme (Project ID: 628043). This work was also funded by a research grant from the Mishima Kaiun Memorial Foundation. We would also like to acknowledge the Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences and Biosciences Division, under award #DE-SC0015662. We would like to thank Jordan Hindes, John Innes Centre, UK for his technical expertise on LPS carbohydrate chemistry, and Adeline Wingertsmann-Cusano for her help with B. subtilis production. We also would like to thank Dr. Tiina Alam?e, University of Tartu, Estonia for our collaboration with using ES levan, and Dr. Harry Gilbert, Dr. Julia-Stefanie Frick, and Dr. Ewa Katzenellenbogen for providing the B. subtilis 168 strain, TLR4 KO cells, and H. alvei LPS, respectively. Funding Information: The authors gratefully acknowledge the support of the Biotechnology and Biological Sciences Research Council (BBSRC); this research was funded by the BBSRC Institute Strategic Programme Grant Food and Health BBS/E/F/00044486 , Food Innovation and Health BB/R012512/1 , Gut Microbes and Health BB/R012490/1 and its constituent project(s), and Molecules from Nature - Products and Pathways BBS/E/J/000PR9790 . Ian D. Young held a BBSRC Doctoral Training Partnership studentship (Project ID: BB/JO14524/1). Norihito Kawasaki would like to thank the Marie-Curie International Incoming Fellowship from the European Union 7th Framework Programme (Project ID: 628043). This work was also funded by a research grant from the Mishima Kaiun Memorial Foundation . We would also like to acknowledge the Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences and Biosciences Division, under award #DE-SC0015662. We would like to thank Jordan Hindes, John Innes Centre, UK for his technical expertise on LPS carbohydrate chemistry, and Adeline Wingertsmann-Cusano for her help with B. subtilis production. We also would like to thank Dr. Tiina Alamäe, University of Tartu, Estonia for our collaboration with using ES levan, and Dr. Harry Gilbert, Dr. Julia-Stefanie Frick, and Dr. Ewa Katzenellenbogen for providing the B. subtilis 168 strain, TLR4 KO cells, and H. alvei LPS, respectively. Publisher Copyright: © 2021 The Authors
Uncontrolled Keywords: fructan,immunomodulatory exopolysaccharide,levan,lipopolysaccharide,tlr4,organic chemistry,polymers and plastics,materials chemistry ,/dk/atira/pure/subjectarea/asjc/1600/1605
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Depositing User: LivePure Connector
Date Deposited: 20 Nov 2021 01:40
Last Modified: 03 Dec 2021 01:43
URI: https://ueaeprints.uea.ac.uk/id/eprint/82248
DOI: 10.1016/j.carbpol.2021.118606

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