Article

Microscale arrays for the profiling of start and stop signals coordinating human-neutrophil swarming

  • Nature Biomedical Engineering 1, Article number: 0094 (2017)
  • doi:10.1038/s41551-017-0094
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Abstract

Neutrophil swarms protect healthy tissues by sealing off sites of infection. In the absence of swarming, microbial invasion of surrounding tissues can result in severe infections. Recent observations in animal models have shown that swarming requires rapid neutrophil responses and well-choreographed neutrophil migration patterns. However, in animal models, physical access to the molecular signals coordinating neutrophil activities during swarming is limited. Here, we report the development and validation of large microscale arrays of zymosan particle clusters for the study of human neutrophils during swarming ex vivo. We characterized the synchronized swarming of human neutrophils under the guidance of neutrophil-released chemokines, and measured the mediators released at different phases of human-neutrophil swarming against targets simulating infections. We found that the network of mediators coordinating human-neutrophil swarming includes start and stop signals, proteolytic enzymes and enzyme inhibitors, as well as modulators of activation of other immune and non-immune cells. We also show that the swarming behaviour of neutrophils from patients following major trauma is deficient and gives rise to smaller swarms than those of neutrophils from healthy individuals.

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Acknowledgements

We thank B. Hamza and J. M. Martel of the BioMEMS Resource Center for help with microfabrication and stimulating discussions. This work was supported by a grant from the National Institute of General Medical Sciences (GM092804) and funding from the Harvard Medical School Tools and Technology Fund. Microfabrication was conducted at the BioMEMS Resource Center at Massachusetts General Hospital, supported by a grant from the National Institute of Biomedical Imaging and Bioengineering (EB002503). Work in the CNS laboratories was supported by the National Institutes of Health (GM095467 and GM38765).

Author information

Affiliations

  1. BioMEMS Resource Center, Department of Surgery, Massachusetts General Hospital, Boston, Massachusetts 02129, USA.

    • Eduardo Reátegui
    • , Fatemeh Jalali
    • , Aimal H. Khankhel
    • , Elisabeth Wong
    • , Hansang Cho
    •  & Daniel Irimia
  2. Massachusetts General Hospital Cancer Center, Boston, Massachusetts 02129, USA.

    • Eduardo Reátegui
    •  & Aimal H. Khankhel
  3. Harvard Medical School, Boston, Massachusetts 02115, USA.

    • Eduardo Reátegui
    • , Hansang Cho
    • , Jarone Lee
    • , Charles N. Serhan
    • , Jesmond Dalli
    • , Hunter Elliott
    •  & Daniel Irimia
  4. Department of Surgery, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.

    • Jarone Lee
  5. Center for Experimental Therapeutics, Department of Anesthesiology, Perioperative, and Pain Medicine, Brigham and Women’s Hospital, Boston, Massachusetts 02115, USA.

    • Charles N. Serhan
    •  & Jesmond Dalli
  6. Image and Data Analysis Core (IDAC), Boston, Massachusetts 02115, USA.

    • Hunter Elliott
  7. Shriners Burns Hospital, Boston, Massachusetts 02114, USA.

    • Daniel Irimia

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Contributions

E.R. and D.I. designed the research; E.R., A.H.K., F.J., H.C. and E.W. designed and prepared the microfluidic devices; E.R. conducted the experiments; E.R., F.J., H.E., J.L. and D.I. analysed the results; C.N.S. and J.D. performed the metabololipidomic analysis; H.E. developed the biophysical model for cell swarming; and E.R. and D.I. prepared the manuscript, with significant contributions from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Daniel Irimia.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary figures, tables and video captions.

Videos

  1. 1.

    Supplementary Video 1

    Human-neutrophil swarming on arrays of clusters of zymosan particles.

  2. 2.

    Supplementary Video 2

    Human-neutrophil swarming on a single cluster of zymosan particles.

  3. 3.

    Supplementary Video 3

    Human neutrophils do not swarm on solitary zymosan particles.

  4. 4.

    Supplementary Video 4

    Human-neutrophil tracking during swarming.

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    Supplementary Video 5

    Human-neutrophil tracking during swarming in the presence of LTB4 receptor.