Letter | Published:

Catapult-like release of mitochondrial DNA by eosinophils contributes to antibacterial defense

Nature Medicine volume 14, pages 949953 (2008) | Download Citation

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Abstract

Although eosinophils are considered useful in defense mechanisms against parasites, their exact function in innate immunity remains unclear. The aim of this study is to better understand the role of eosinophils within the gastrointestinal immune system. We show here that lipopolysaccharide from Gram-negative bacteria activates interleukin-5 (IL-5)- or interferon-γ–primed eosinophils to release mitochondrial DNA in a reactive oxygen species–dependent manner, but independent of eosinophil death. Notably, the process of DNA release occurs rapidly in a catapult-like manner—in less than one second. In the extracellular space, the mitochondrial DNA and the granule proteins form extracellular structures able to bind and kill bacteria both in vitro and under inflammatory conditions in vivo. Moreover, after cecal ligation and puncture, Il5-transgenic but not wild-type mice show intestinal eosinophil infiltration and extracellular DNA deposition in association with protection against microbial sepsis. These data suggest a previously undescribed mechanism of eosinophil-mediated innate immune responses that might be crucial for maintaining the intestinal barrier function after inflammation-associated epithelial cell damage, preventing the host from uncontrolled invasion of bacteria.

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Acknowledgements

We thank M. Kohler for critical review of the confocal microscopy and imaging data. We also thank D. Simon for histological examination of the mouse tissue sections. This work was supported by the Swiss National Science Foundation (grants 310000-112078 and 310000-107526) and Stanley Thomas Johnson Foundation, Bern, Switzerland.

Author information

Affiliations

  1. Institute of Pharmacology, University of Bern, Friedbühlstrasse 49, CH-3010 Bern, Switzerland.

    • Shida Yousefi
    • , Nicola Andina
    • , Evelyne Kozlowski
    • , Inès Schmid
    •  & Hans-Uwe Simon
  2. Division of Pulmonary and Critical Care, Oregon Health and Sciences University, 3181 Southwest Sam Jackson Park Road, Portland, Oregon 97239, USA.

    • Jeffrey A Gold
    •  & Ann M Kelly
  3. Department of Biochemistry and Molecular Biology, Mayo Clinic Arizona, 13400 East Shea Boulevard, Scottsdale, Arizona 85259, USA.

    • James J Lee
  4. Department of Gastroenterology, Kantonsspital Olten, Römerstrasse 7, CH-4600 Olten, Switzerland.

    • Alex Straumann
  5. University Children's Hospital, Steinwiesstrasse 75, CH-8032 Zurich, Switzerland.

    • Janine Reichenbach
  6. Department of Dermatology, University of Utah, 30 North 1900 East, Salt Lake City, Utah 84132-2409, USA.

    • Gerald J Gleich

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Contributions

S.Y. developed techniques and conducted most of the experiments. J.A.G., N.A., A.M.K., E.K. and I.S. performed experiments. A.S. and J.R. cared for the involved human subjects and provided clinical samples. J.J.L. provided mice and helped design the mouse experiments. G.J.G. provided reagents and suggestions on the project. S.Y. and H.-U.S. designed the project. H.-U.S. supervised the project and wrote most of the manuscript.

Corresponding authors

Correspondence to Shida Yousefi or Hans-Uwe Simon.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figs. 1–6 and Suppementary Methods

Videos

  1. 1.

    Supplementary Movie 1 Part 1

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vivo conditions (Crohn's disease). DNA was stained with PI and ECP with a specific antibody. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 10 (left) and 15 (right) slices per stack in order to show extracellular DNA in their full extension (location is indicated by arrows).

  2. 2.

    Supplementary Movie 1 Part 2

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vivo conditions (Crohn's disease). DNA was stained with PI and ECP with a specific antibody. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 10 (left) and 15 (right) slices per stack in order to show extracellular DNA in their full extension (location is indicated by arrows).

  3. 3.

    Supplementary Movie 2 Part 1

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vivo conditions (Schistosoma infection). DNA was stained with PI and ECP/MBP with specific antibodies. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 50 (left) and 20 (right) slices per stack in order to show extracellular DNA in their full extension (the location of areas with large extracellular DNA deposition are indicated by dashed circles).

  4. 4.

    Supplementary Movie 2 Part 2

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vivo conditions (Schistosoma infection). DNA was stained with PI and ECP/MBP with specific antibodies. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 50 (left) and 20 (right) slices per stack in order to show extracellular DNA in their full extension (the location of areas with large extracellular DNA deposition are indicated by dashed circles).

  5. 5.

    Supplementary Movie 3 Part 1

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vitro conditions. Highly purified blood eosinophils were stimulated with IL-5/LPS. DNA was stained with PI and granule proteins with specific antibodies. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 10 slices per stack in order to show extracellular DNA in their full extension.

  6. 6.

    Supplementary Movie 3 Part 2

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vitro conditions. Highly purified blood eosinophils were stimulated with IL-5/LPS. DNA was stained with PI and granule proteins with specific antibodies. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 10 slices per stack in order to show extracellular DNA in their full extension.

  7. 7.

    Supplementary Movie 3 Part 3

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vitro conditions. Highly purified blood eosinophils were stimulated with IL-5/LPS. DNA was stained with PI and granule proteins with specific antibodies. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 10 slices per stack in order to show extracellular DNA in their full extension.

  8. 8.

    Supplementary Movie 3 Part 4

    Demonstration of extracellular DNA and eosinophil granule protein deposition under in vitro conditions. Highly purified blood eosinophils were stimulated with IL-5/LPS. DNA was stained with PI and granule proteins with specific antibodies. With an interval of 0.1 μm, slices were taken throughout the z axis (z stacks) with 10 slices per stack in order to show extracellular DNA in their full extension.

  9. 9.

    Supplementary Movie 4

    Release of DNA from activated eosinophils as assessed by live cell imaging analysis. DNA was stained with SYTO 13. One particular eosinophil (arrow) was followed in a time-dependent manner. Release of DNA occurred with less than 1 second.

  10. 10.

    Supplementary Movie 5

    Release of DNA from activated eosinophils as assessed by live cell imaging analysis. DNA was stained with SYTO 13. It appears that the DNA is released from perinuclear but not nuclear structures.

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DOI

https://doi.org/10.1038/nm.1855

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