Article | Published:

Nod2-mediated recognition of the microbiota is critical for mucosal adjuvant activity of cholera toxin

Nature Medicine volume 22, pages 524530 (2016) | Download Citation

  • A Corrigendum to this article was published on 04 August 2016

This article has been updated

Abstract

Cholera toxin (CT) is a potent adjuvant for inducing mucosal immune responses. However, the mechanism by which CT induces adjuvant activity remains unclear. Here we show that the microbiota is critical for inducing antigen-specific IgG production after intranasal immunization. After mucosal vaccination with CT, both antibiotic-treated and germ-free (GF) mice had reduced amounts of antigen-specific IgG, smaller recall-stimulated cytokine responses, impaired follicular helper T (TFH) cell responses and reduced numbers of plasma cells. Recognition of symbiotic bacteria via the nucleotide-binding oligomerization domain containing 2 (Nod2) sensor in cells that express the integrin CD11c (encoded by Itgax) was required for the adjuvanticity of CT. Reconstitution of GF mice with a Nod2 agonist or monocolonization with Staphylococcus sciuri, which has high Nod2-stimulatory activity, was sufficient to promote robust CT adjuvant activity, whereas bacteria with low Nod2-stimulatory activity did not. Mechanistically, CT enhanced Nod2-mediated cytokine production in dendritic cells via intracellular cyclic AMP. These results show a role for the microbiota and the intracellular receptor Nod2 in promoting the mucosal adjuvant activity of CT.

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Change history

  • 26 May 2016

    In the version of this article initially published, Mathias Chamaillard was inadvertently omitted from the list of authors and from the Author Contributions section. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

This work was supported by US National Institutes of Health (NIH) grant R01 DK61707 (G.N.), the University of Michigan's Cancer Center Support grant 5 P30 CA46592 (G.N.), the DFG Cluster of Excellence 'Inflammation at Interfaces' (P.R.) and the BMBF grant TP5 (P.R.). We thank M. Zeng for critical review of the manuscript, L. Haynes for animal husbandry and the University of Michigan Germ-Free Animal Core Facility and Host Microbiome Initiative (G.N.) for support.

Author information

Affiliations

  1. Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Donghyun Kim
    • , Yun-Gi Kim
    • , Sang-Uk Seo
    • , Dong-Jae Kim
    • , Naohiro Inohara
    •  & Gabriel Núñez
  2. Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Donghyun Kim
    • , Yun-Gi Kim
    • , Sang-Uk Seo
    • , Dong-Jae Kim
    • , Naohiro Inohara
    •  & Gabriel Núñez
  3. Department of Internal Medicine, Division of Gastroenterology, University of Michigan Medical School, Ann Arbor, Michigan, USA.

    • Nobuhiko Kamada
  4. Department of Immunology, University of Toronto, Toronto, Ontario, Canada.

    • Dave Prescott
    •  & Dana J Philpott
  5. Center for Infection and Immunity of Lille, Institute Pasteur of Lille, Lille, France.

    • Mathias Chamaillard
  6. Institute of Clinical Molecular Biology, University of Kiel, Kiel, Germany.

    • Philip Rosenstiel

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Contributions

D.K., Y.-G.K. and G.N. conceived the study. D.K. performed most of the experiments. Y.-G.K. performed several experiments. S.-U.S. and D.-J.K. helped with experiments. D.P., D.J.P. and P.R. generated and performed initial characterization of Itgax-Cre;Nod2fl/fl mice. M.C. provided critical materials. N.K., D.J.P., P.R. and N.I. helped in the design of several experiments and provided critical advice. D.K., Y.-G.K. and G.N. wrote the manuscript, with contributions from all of the authors.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Yun-Gi Kim or Gabriel Núñez.

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DOI

https://doi.org/10.1038/nm.4075

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