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Illuminating vital surface molecules of symbionts in health and disease

Nature Microbiology volume 2, Article number: 17099 (2017) Cite this article

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Abstract

The immunomodulatory surface molecules of commensal and pathogenic bacteria are critical to microorganisms' survival and the host's response1,2. Recent studies have highlighted the unique and important responses elicited by commensal-derived surface macromolecules3–5. However, the technology available to track these molecules in host cells and tissues remains primitive. We report, here, an interdisciplinary approach that uses metabolic labelling combined with bioorthogonal click chemistry (that is, reactions performed in living organisms)6 to specifically tag up to three prominent surface immunomodulatory macromolecules—peptidoglycan, lipopolysaccharide and capsular polysaccharide—either simultaneously or individually in live anaerobic commensal bacteria. Importantly, the peptidoglycan labelling enables, for the first time, the specific labelling of live endogenous, anaerobic bacteria within the mammalian host. This approach has allowed us to image and track the path of labelled surface molecules from live, luminal bacteria into specific intestinal immune cells in the living murine host during health and disease. The chemical labelling of three specific macromolecules within a live organism offers the potential for in-depth visualization of host–pathogen interactions.

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Figure 1: Fluorescent d-amino acid labels PGN in commensal bacteria.
Figure 2: Use of HADA-labelled PGN to track live commensals in the host.
Figure 3: Simultaneous labelling of three cell surface molecules in commensal bacteria.
Figure 4: Simultaneous labelling of three cell surface molecules in commensal bacteria.

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Acknowledgements

The authors acknowledge the Harvard Medical School Center for Immune Imaging for providing instrumentation and aid for two-photon microscopy. The authors thank N. Geva-Zatorsky and F. Gazzaniga for materials, expertise and discussion, D. Erturk-Hasdemir and N. Okan for aid in cell culture and isolation of bone marrow macrophages, and C. Hudak for discussion and manuscript critique. This work was funded by a grant from the US Department of Defense (W81XWH-15-1-0368) and was supported in part by the US National Institutes of Health (grants PO1 AI1112521, RO1 AI111595 (to U.H.v.A.) and 5T32 HL066987 (to D.A.)). Additional support to U.H.v.A. was provided by the Ragon Institute at MGH, MIT and Harvard. J.E.H. was supported by the Cancer Research Institute Irvington Fellowship Program.

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Authors and Affiliations

  1. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Jason E. Hudak, David Alvarez, Ashwin Skelly, Ulrich H. von Andrian & Dennis L. Kasper

  2. The Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology and Harvard, Cambridge, 02139, Massachusetts, USA

    Ulrich H. von Andrian

Authors
  1. Jason E. Hudak
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  2. David Alvarez
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  4. Ulrich H. von Andrian
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Contributions

J.E.H. designed the experiments, analysed the data and wrote the manuscript with help from D.A. and A.S. D.A. provided expertise in two-photon intravital microscopy. D.L.K. supervised the study, edited the manuscript and provided helpful comments, with assistance from U.H.v.A.

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Correspondence to Dennis L. Kasper.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Figures 1–13, Supplementary Notes and Supplementary References. (PDF 18026 kb)

Supplementary Video 1

Intravital two-photon microscopy of the commensal microflora in the mouse colon. (MOV 12295 kb)

Supplementary Video 2

Intravital two-photon microscopy of the commensal microflora in the mouse colon. (MOV 11091 kb)

Supplementary Video 3

Intravital two-photon microscopy of the commensal microbe B. vulgatus in the mouse small intestine. (MOV 4731 kb)

Supplementary Video 4

Intravital two-photon microscopy of the commensal microbe B. vulgatus in the mouse small intestine. (MOV 6610 kb)

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Hudak, J., Alvarez, D., Skelly, A. et al. Illuminating vital surface molecules of symbionts in health and disease. Nat Microbiol 2, 17099 (2017). https://doi.org/10.1038/nmicrobiol.2017.99

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