Topologically correct synthetic reconstruction of pathogen social behavior found during Yersinia growth in deep tissue sites

  1. Stacie A Clark
  2. Derek Thibault
  3. Lauren M Shull
  4. Kimberly M Davis
  5. Emily Aunins
  6. Tim van Opijnen  Is a corresponding author
  7. Ralph Isberg  Is a corresponding author
  1. Tufts University, United States
  2. Boston College, United States
  3. Johns Hopkins Bloomberg School of Public Health, United States

Abstract

Within deep tissue sites, extracellular bacterial pathogens often replicate in clusters that are surrounded by immune cells. Disease is modulated by interbacterial interactions as well as bacterial-host cell interactions resulting in microbial growth, phagocytic attack and secretion of host antimicrobial factors. To overcome the limited ability to manipulate these infection sites, we established a system for Yersinia pseudotuberculosis (Yptb) growth in microfluidics-driven microdroplets that regenerates microbial social behavior in tissues. Chemical generation of nitric oxide (NO) in the absence of immune cells was sufficient to reconstruct microbial social behavior, as witnessed by expression of the NO-inactivating protein Hmp on the extreme periphery of microcolonies, mimicking spatial regulation in tissues. Similarly, activated macrophages that expressed inducible NO synthase (iNOS) drove peripheral expression of Hmp, allowing regeneration of social behavior observed in tissues. These results argue that topologically correct microbial tissue growth and associated social behavior can be reconstructed in culture.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Scripts described have been deposited with GitHub and the appropriate link is provided in manuscript.

Article and author information

Author details

  1. Stacie A Clark

    Molecular Biology and Microbioloty, Tufts University, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Derek Thibault

    Biology, Boston College, Chestnut Hill, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Lauren M Shull

    Department of Molecular Biology and Microbiology, Tufts University, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Kimberly M Davis

    Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Emily Aunins

    Molecular Biology and Microbiology, Tufts University, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Tim van Opijnen

    Biology, Boston College, Chestnut Hill, United States
    For correspondence
    vanopijn@bc.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6895-6795
  7. Ralph Isberg

    Molecular Biology and Microbiology, Tufts University, Boston, United States
    For correspondence
    ralph.isberg@tufts.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8330-3554

Funding

National Institute of Allergy and Infectious Diseases (R01 AI110684)

  • Ralph Isberg

National Institute of Allergy and Infectious Diseases (R21 151593)

  • Ralph Isberg

National Institute of Allergy and Infectious Diseases (U01 AI124302)

  • Tim van Opijnen

National Institute of Allergy and Infectious Diseases (R01 AI110724)

  • Tim van Opijnen

National Institute of General Medical Sciences (T32 TM007310)

  • Stacie A Clark

National Institute of General Medical Sciences (T32 TM007310)

  • Derek Thibault

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Sophie Helaine, Imperial College London, United Kingdom

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#B2016-21 and B2019-03 ) of Tufts University. No surgeries were performed.

Version history

  1. Received: April 21, 2020
  2. Accepted: June 9, 2020
  3. Accepted Manuscript published: June 16, 2020 (version 1)
  4. Version of Record published: June 25, 2020 (version 2)

Copyright

© 2020, Clark et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Stacie A Clark
  2. Derek Thibault
  3. Lauren M Shull
  4. Kimberly M Davis
  5. Emily Aunins
  6. Tim van Opijnen
  7. Ralph Isberg
(2020)
Topologically correct synthetic reconstruction of pathogen social behavior found during Yersinia growth in deep tissue sites
eLife 9:e58106.
https://doi.org/10.7554/eLife.58106

Share this article

https://doi.org/10.7554/eLife.58106

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