Abstract
Background
Otitis media (OM), a prevalent pediatric infectious disease, is mainly caused by Streptococcus pneumoniae (S.pn). Neutrophil extracellular traps (NETs), a novel antimicrobial strategy, were reported in 2004. We found that NETs formed in the middle ear with acute otitis media (AOM) induced by S.pn. However, the mechanisms of NETs formation are not entirely clear.
Methods
We stimulated neutrophils isolated from mouse bone marrow with S.pn clinical stain 19F in vitro, and established mouse model of AOM via transbullar injection with S.pn. NETs formation, reactive oxygen species (ROS) production, autophagy activation and bacterial load were analyzed in TLR4−/− and wild-type neutrophils stimulated in vitro with S.pn and in vivo during AOM.
Results
We found that autophagy and ROS were required for S.pn-induced NETs formation. Moreover, TLR4 partly mediated NETs formation in response to S.pn in vitro and in vivo during AOM. We also showed that attenuated NETs formation in TLR4−/− neutrophils correlated with an impaired ROS production and autophagy activation in vitro and in vivo. In addition, both the in vivo and in vitro-produced NETs were able to engulf and kill S.pn.
Conclusions
TLR4 regulates ROS and autophagy to control NETs formation against S.pn in the course of AOM.
Impact
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S.pn can induce NETs formation in vitro and in vivo; TLR4 regulates NETs formation by ROS and autophagy; NETs contribute to the clearance of bacteria in acute otitis media.
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In this study, we firstly found that autophagy and ROS were required for S.pn-induced NETs formation in the model of acute otitis media (AOM). And to some extent, TLR4 mediated NETs formation during AOM.
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Our research might provide a potential strategy for the treatment of otitis media.
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Acknowledgements
This work was supported by the National Natural Science Foundation Grants of China (No. csfc81373151) and Natural Science Foundation Project of CQCSTC (Nos. cstc2012jjA0035, cstc2018jcyjAX0257).
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Y.H. and Y.D. conceived of and designed the research. Y.D., C.J., Y.Z., Q.H., X.Z. and Z.D. performed the experiments and analyzed the data. Y.H., Z.D. and Y.D. wrote the manuscript. R.A. and Y.Y. interpreted the data.
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Dong, Y., Jin, C., Ding, Z. et al. TLR4 regulates ROS and autophagy to control neutrophil extracellular traps formation against Streptococcus pneumoniae in acute otitis media. Pediatr Res 89, 785–794 (2021). https://doi.org/10.1038/s41390-020-0964-9
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DOI: https://doi.org/10.1038/s41390-020-0964-9
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