Colistin resistance mutations in phoQ can sensitize Klebsiella pneumoniae to IgM-mediated complement killing

Sjors P A van der Lans; Manon Janet-Maitre; Frerich M Masson; Kimberly A Walker; Dennis J Doorduijn; Axel B Janssen; Willem van Schaik; Ina Attrée; Suzan H M Rooijakkers; Bart W Bardoel

doi:10.1038/s41598-023-39613-5

Colistin resistance mutations in phoQ can sensitize Klebsiella pneumoniae to IgM-mediated complement killing

Sjors P A van der Lans, Manon Janet-Maitre, Frerich M Masson, Kimberly A Walker, Dennis J Doorduijn, Axel B Janssen, Willem van Schaik, Ina Attrée, Suzan H M Rooijakkers, Bart W Bardoel

Microbiology and Infection

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Abstract

Due to multi-drug resistance, physicians increasingly use the last-resort antibiotic colistin to treat infections with the Gram-negative bacterium Klebsiella pneumoniae. Unfortunately, K. pneumoniae can also develop colistin resistance. Interestingly, colistin resistance has dual effects on bacterial clearance by the immune system. While it increases resistance to antimicrobial peptides, colistin resistance has been reported to sensitize certain bacteria for killing by human serum. Here we investigate the mechanisms underlying this increased serum sensitivity, focusing on human complement which kills Gram-negatives via membrane attack complex (MAC) pores. Using in vitro evolved colistin resistant strains and a fluorescent MAC-mediated permeabilization assay, we showed that two of the three tested colistin resistant strains, Kp209_CSTR and Kp257_CSTR, were sensitized to MAC. Transcriptomic and mechanistic analyses focusing on Kp209_CSTR revealed that a mutation in the phoQ gene locked PhoQ in an active state, making Kp209_CSTR colistin resistant and MAC sensitive. Detailed immunological assays showed that complement activation on Kp209_CSTR in human serum required specific IgM antibodies that bound Kp209_CSTR but did not recognize the wild-type strain. Together, our results show that developing colistin resistance affected recognition of Kp209_CSTR and its killing by the immune system.

Original language	English
Article number	12618
Number of pages	14
Journal	Scientific Reports
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41598-023-39613-5
Publication status	Published - 3 Aug 2023

Bibliographical note

© 2023. Springer Nature Limited.

Funding
This work was supported by the Netherlands Organization for Scientific Research (NWO) through a TTW-NACTAR Grant #16442 (to SvdL and SHMR) and the European Union's Horizon 2020 research programs 787 H2020-EU-ITN-EJD (CORVOS #860044 to FM and SHMR). The funders had no role in the study design, data collection and analysis, or preparation of the manuscript.

Keywords

Humans
Colistin/pharmacology
Klebsiella pneumoniae/genetics
Bacterial Proteins/pharmacology
Drug Resistance, Bacterial/genetics
Anti-Bacterial Agents/pharmacology
Mutation
Immunoglobulin M/genetics
Klebsiella Infections/drug therapy
Microbial Sensitivity Tests

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van der Lans, S. P. A., Janet-Maitre, M., Masson, F. M., Walker, K. A., Doorduijn, D. J., Janssen, A. B., van Schaik, W., Attrée, I., Rooijakkers, S. H. M., & Bardoel, B. W. (2023). Colistin resistance mutations in phoQ can sensitize Klebsiella pneumoniae to IgM-mediated complement killing. Scientific Reports, 13(1), Article 12618. https://doi.org/10.1038/s41598-023-39613-5

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abstract = "Due to multi-drug resistance, physicians increasingly use the last-resort antibiotic colistin to treat infections with the Gram-negative bacterium Klebsiella pneumoniae. Unfortunately, K. pneumoniae can also develop colistin resistance. Interestingly, colistin resistance has dual effects on bacterial clearance by the immune system. While it increases resistance to antimicrobial peptides, colistin resistance has been reported to sensitize certain bacteria for killing by human serum. Here we investigate the mechanisms underlying this increased serum sensitivity, focusing on human complement which kills Gram-negatives via membrane attack complex (MAC) pores. Using in vitro evolved colistin resistant strains and a fluorescent MAC-mediated permeabilization assay, we showed that two of the three tested colistin resistant strains, Kp209_CSTR and Kp257_CSTR, were sensitized to MAC. Transcriptomic and mechanistic analyses focusing on Kp209_CSTR revealed that a mutation in the phoQ gene locked PhoQ in an active state, making Kp209_CSTR colistin resistant and MAC sensitive. Detailed immunological assays showed that complement activation on Kp209_CSTR in human serum required specific IgM antibodies that bound Kp209_CSTR but did not recognize the wild-type strain. Together, our results show that developing colistin resistance affected recognition of Kp209_CSTR and its killing by the immune system.",

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AU - van der Lans, Sjors P A

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AU - Walker, Kimberly A

AU - Doorduijn, Dennis J

AU - Janssen, Axel B

AU - van Schaik, Willem

AU - Attrée, Ina

AU - Rooijakkers, Suzan H M

AU - Bardoel, Bart W

N1 - © 2023. Springer Nature Limited. Funding This work was supported by the Netherlands Organization for Scientific Research (NWO) through a TTW-NACTAR Grant #16442 (to SvdL and SHMR) and the European Union's Horizon 2020 research programs 787 H2020-EU-ITN-EJD (CORVOS #860044 to FM and SHMR). The funders had no role in the study design, data collection and analysis, or preparation of the manuscript.

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N2 - Due to multi-drug resistance, physicians increasingly use the last-resort antibiotic colistin to treat infections with the Gram-negative bacterium Klebsiella pneumoniae. Unfortunately, K. pneumoniae can also develop colistin resistance. Interestingly, colistin resistance has dual effects on bacterial clearance by the immune system. While it increases resistance to antimicrobial peptides, colistin resistance has been reported to sensitize certain bacteria for killing by human serum. Here we investigate the mechanisms underlying this increased serum sensitivity, focusing on human complement which kills Gram-negatives via membrane attack complex (MAC) pores. Using in vitro evolved colistin resistant strains and a fluorescent MAC-mediated permeabilization assay, we showed that two of the three tested colistin resistant strains, Kp209_CSTR and Kp257_CSTR, were sensitized to MAC. Transcriptomic and mechanistic analyses focusing on Kp209_CSTR revealed that a mutation in the phoQ gene locked PhoQ in an active state, making Kp209_CSTR colistin resistant and MAC sensitive. Detailed immunological assays showed that complement activation on Kp209_CSTR in human serum required specific IgM antibodies that bound Kp209_CSTR but did not recognize the wild-type strain. Together, our results show that developing colistin resistance affected recognition of Kp209_CSTR and its killing by the immune system.

AB - Due to multi-drug resistance, physicians increasingly use the last-resort antibiotic colistin to treat infections with the Gram-negative bacterium Klebsiella pneumoniae. Unfortunately, K. pneumoniae can also develop colistin resistance. Interestingly, colistin resistance has dual effects on bacterial clearance by the immune system. While it increases resistance to antimicrobial peptides, colistin resistance has been reported to sensitize certain bacteria for killing by human serum. Here we investigate the mechanisms underlying this increased serum sensitivity, focusing on human complement which kills Gram-negatives via membrane attack complex (MAC) pores. Using in vitro evolved colistin resistant strains and a fluorescent MAC-mediated permeabilization assay, we showed that two of the three tested colistin resistant strains, Kp209_CSTR and Kp257_CSTR, were sensitized to MAC. Transcriptomic and mechanistic analyses focusing on Kp209_CSTR revealed that a mutation in the phoQ gene locked PhoQ in an active state, making Kp209_CSTR colistin resistant and MAC sensitive. Detailed immunological assays showed that complement activation on Kp209_CSTR in human serum required specific IgM antibodies that bound Kp209_CSTR but did not recognize the wild-type strain. Together, our results show that developing colistin resistance affected recognition of Kp209_CSTR and its killing by the immune system.

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KW - Mutation

KW - Immunoglobulin M/genetics

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DO - 10.1038/s41598-023-39613-5

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C2 - 37537263

SN - 2045-2322

VL - 13

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 12618

ER -

Colistin resistance mutations in phoQ can sensitize Klebsiella pneumoniae to IgM-mediated complement killing

Abstract

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Keywords

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