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Synergistic bactericidal activities of tobramycin with ciprofloxacin and azithromycin against Klebsiella pneumoniae

Abstract

Trans-translation is a unique bacterial ribosome rescue system that plays important roles in the tolerance to environmental stresses. It is composed of an ssrA-encoded tmRNA and a protein SmpB. In this study, we examined the role of trans-translation in antibiotic tolerance in Klebsiella pneumoniae and explored whether the inhibition of this mechanism could enhance the bactericidal activities of antibiotics. We found that deletion of the ssrA gene reduced the survival of K. pneumoniae after treatment with kanamycin, tobramycin, azithromycin, and ciprofloxacin, indicating an important role of the trans-translation in bacterial antibiotic tolerance. By using a modified ssrA gene with a 6×His tag we demonstrated that tobramycin suppressed the azithromycin and ciprofloxacin-elicited activation of trans-translation. The results were further confirmed with a trans-translation reporter system that is composed of a normal mCherry gene and a gfp gene without the stop codon. Compared to each individual antibiotic, combination of tobramycin with azithromycin or ciprofloxacin synergistically enhanced the killing activities against planktonic K. pneumoniae cells and improved bacterial clearance in a murine cutaneous abscess infection model. In addition, the combination of tobramycin and ciprofloxacin increased the bactericidal activities against biofilm-associated cells. Overall, our results suggest that the combination of tobramycin with azithromycin or ciprofloxacin is a promising strategy in combating K. pneumoniae infections.

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Acknowledgements

We would like to thank Dr. Robert E. W. Hancock at the Centre for Microbial Diseases and Immunity Research, Department of Microbiology and Immunology, University of British Columbia for the teaching of the mouse cutaneous abscess model. We would like to thank Dr. Quanjiang Ji at the School of Physical Science and Technology, ShanghaiTech University for kindly providing the K. pneumoniae CRISPR system. This work was supported by National Key Research and Development Project of China (2021YFE0101700, 2017YFE0125600), National Science Foundation of China (31970680, 31970179 and 31870130), Fundamental Research Funds for the Central Universities, Nankai University (63201093), and the Tianjin Municipal Science and Technology Commission (19JCYBJC24700). The funders had no role in study design, data collection, and interpretation, or the decision to submit the work for publication.

Author contributions

W.W., H.R., and D.P. conceived and designed the experiments. H.R., Y.Z., J.Z., C.X., Z.F., X.P., and S.L. performed the experiments. H.R., Y.J., F.B., Z.C., D.P., and W.W. analyzed the data. Y.Z., S.C., J.X., P.W., Yh.Z., G.Z., H.L. provided and characterized the clinical isolates. H.R., D.P., and W.W. wrote the paper.

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Correspondence to Weihui Wu.

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The animal experiments were conducted following the national guidelines on the use of animals in research. The protocol was approved by the Animal Care and Use Committee of the College of Life Sciences, Nankai University (permission number: NK-04-2012).

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Ren, H., Zhang, J., Zhou, J. et al. Synergistic bactericidal activities of tobramycin with ciprofloxacin and azithromycin against Klebsiella pneumoniae. J Antibiot 74, 528–537 (2021). https://doi.org/10.1038/s41429-021-00427-0

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