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⇱ Respiratory protein–generated reactive oxygen species as an antimicrobial strategy | Nature Immunology

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Abstract

The evolution of the host-pathogen relationship comprises a series of invasive-defensive tactics elicited by both participants. The stereotype is that the antimicrobial immune response requires multistep processes. Little is known about the primordial immunosurveillance system, which probably has components that directly link sensors and effectors. Here we found that the respiratory proteins of both the horseshoe crab and human were directly activated by microbial proteases and were enhanced by pathogen-associated molecular patterns, resulting in the production of more reactive oxygen species. Hemolytic virulent pathogens, which produce proteases as invasive factors, are more susceptible to this killing mechanism. This 'shortcut' antimicrobial strategy represents a fundamental and universal mode of immunosurveillance, which has been in existence since before the split of protostomes and deuterostomes and still persists today.

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Figure 1: Activation of HMC-PPO to PO by microbial proteases and PAMPs.
Figure 2: In vitro antimicrobial action of activated PO.
Figure 3: PO triggered by microbial protease contributes to in vivo bacterial clearance.
Figure 4: Pseudoperoxidase activity of metHb is triggered synergistically by microbial proteases and PAMPs.
Figure 5: Functional evaluation of the metHb-mediated ROS production in RBC by various bacteria.

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Acknowledgements

We thank B.H. Iglewski (University of Rochester School of Medicine and Dentistry) for P. aeruginosa strains PAO-Iglewski and PAO-B1A1; S. Arvidson (Karolinska University, Sweden) for S. aureus strains PC1839 and AK3; L. Zhang (Institute of Molecular and Cell Biology, Singapore) for plasmid pDSK-GFP; A. Cheung (Dartmouth Medical School) for plasmid pALC1420; H.C. Ng for bacterial isolation and technical support; S. Leptihn for suggestions on bacterial identification by 16S rRNA sequencing; and B. Halliwell, A. Le Saux and P.M.L. Ng for critical reading of the manuscript. Supported by the Agency for Science Technology and Research (Biomedical Research Council), the Academic Research Fund of the Ministry of Education, and the Faculty Research Committee of the National University of Singapore.

Author information

Author notes

  1. Bow Ho and Jeak Ling Ding: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Sciences, National University of Singapore, Singapore, 117543

    Naxin Jiang & Jeak Ling Ding

  2. School of Biological Sciences, Nanyang Technological University, Singapore, 637551

    Nguan Soon Tan

  3. Department of Microbiology, National University of Singapore, Singapore, 117597

    Bow Ho

Authors
  1. Naxin Jiang
  2. Nguan Soon Tan
  3. Bow Ho
  4. Jeak Ling Ding

Contributions

N.J. designed and did the experiments and prepared the manuscript; N.S.T. provided some suggestions, helped with real-time fluorescence imaging and manuscript preparation; B.H. provided expertise in microbiology, designed and did some of the experiments and participated in manuscript preparation; J.L.D. conceived the ideas, designed the biochemical, cell and molecular biology experiments, and supervised the studies and preparation of the manuscript.

Corresponding author

Correspondence to Jeak Ling Ding.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures (download PDF )

Supplementary Figures 1–11, Methods and Text (PDF 14224 kb)

Supplementary Video 1 (download MOV )

The real-time imaging of the bacterial clearance elicited by PO-catalyzed production of quinone using PAO-Iglewski, the wild type Pseudomonas aeruginosa strain which produces extracellular protease. (MOV 993 kb)

Supplementary Video 2 (download MOV )

The real-time imaging of the bacterial clearance elicited by PO-catalyzed production of quinone using PAO-B1A1, the Pseudomonas aeruginosa mutant strain which is protease-deficient. (MOV 1183 kb)

Supplementary Video 3 (download MOV )

The real-time imaging of the bacterial clearance elicited by PO-catalyzed production of quinone using PAO-Iglewski in the presence of phenylthiourea, a specific inhibitor of the phenoloxidase. (MOV 803 kb)

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Jiang, N., Tan, N., Ho, B. et al. Respiratory protein–generated reactive oxygen species as an antimicrobial strategy. Nat Immunol 8, 1114–1122 (2007). https://doi.org/10.1038/ni1501

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