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Neutrophil extracellular traps may have a dual role in Pseudomonas aeruginosa keratitis

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European Journal of Clinical Microbiology & Infectious Diseases Aims and scope Submit manuscript

Abstract

Pseudomonas aeruginosa (P. aeruginosa) keratitis is a sight-threatening and rapidly progressive corneal disease. Neutrophils and neutrophil extracellular traps (NETs) are widely thought to play a vital role in hosts’ immune defenses against bacteria, such as P. aeruginosa. The present study aimed to investigate the dynamics of the formation and the role of NETs in P. aeruginosa keratitis. First, scratched corneas of mice models were treated with 1 × 108 colony-forming units (CFU)/ml of P. aeruginosa suspension or normal saline (NS). Second, after 48 h postinfection, the infected corneas were treated with TobraDex, Tobrex, 0.1% dexamethasone, or NS four times a day, respectively. Clinical examination, hematoxylin and eosin (H&E) staining, immunofluorescence staining, scanning electron microscopy, and bacterial burden testing were performed on the corneas. Tobrex reduced neutrophil infiltration and corneal P. aeruginosa burden. Dexamethasone reduced NETs, bacterial burden, and severe neutrophil infiltration. TobraDex produced a greater reduction in the amount of neutrophils, NETs, and bacterial burden and the results of Tobrex-treated group were between them. These findings corresponded with the clinical findings that TobraDex- and Tobrex-treated mice exhibited slight corneal damage, while dexamethasone-treated mice exhibited very severe corneal damage. Cumulatively, our data suggest that NETs may play a dual role of infection control and corneal damage in P. aeruginosa keratitis. Furthermore, combination treatment targeting NET formation and bacteria may serve as a way of improving the clinical outcomes of bacterial keratitis.

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Data availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

CFU:

colony-forming units

H&E:

hematoxylin and eosin

MPO:

myeloperoxidase

NETs:

neutrophil extracellular traps

NS:

normal saline

P. aeruginosa :

Pseudomonas aeruginosa

PBS:

phosphate-buffered saline

PFA:

paraformaldehyde

SEM:

scanning electron microscope

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Funding

This study was funded by the National Natural Science Foundation of China (Grant number: 81270974; 81870624) and major science and technology projects of Zhejiang province (Grant number: 2017C03046).

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Authors and Affiliations

  1. Eye Center, Second Affiliated Hospital, School of Medicine, Zhejiang University, 88 Jiefang Road, Hangzhou, 310009, China

    Binbin Zhu, Lu Zhang, Kelan Yuan, Xiaodan Huang, Renjian Hu & Xiuming Jin

Authors
  1. Binbin Zhu
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  2. Lu Zhang
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  3. Kelan Yuan
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  4. Xiaodan Huang
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  5. Renjian Hu
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  6. Xiuming Jin
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Contributions

BZ, XH, and XJ initially designed the concept of this work, BZ, LZ, YK, and RH collected and analyzed the data, BZ and XJ wrote the manuscript, and BZ and LZ revised the manuscript. All authors have read and approved the manuscript.

Corresponding author

Correspondence to Xiuming Jin.

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Competing interests

The authors declare that they have no competing interests.

Ethics approval

This study was approved by the Ethics Committee of the Second Affiliated Hospital at the School of Medicine, Zhejiang University, China. Procedures involving animals were in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research.

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ESM 1

Supplementary Figure: Purified human neutrophils were stained using SytoxGreen to label the DNA in vitro. A: Human neutrophils were stimulated by P. aeruginosa. B: Human neutrophils were pretreated with dexamethasone for 30 minutes and then stimulated with P. aeruginosa. C: As controls, human neutrophils were untreated with P. aeruginosa or dexamethasone. Magnification, ×200. (JPG 41 kb)

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Zhu, B., Zhang, L., Yuan, K. et al. Neutrophil extracellular traps may have a dual role in Pseudomonas aeruginosa keratitis. Eur J Clin Microbiol Infect Dis 40, 169–180 (2021). https://doi.org/10.1007/s10096-020-04023-2

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  • DOI: https://doi.org/10.1007/s10096-020-04023-2

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