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Mycobacteria form viable cell wall-deficient cells that are undetectable by conventional diagnostics

Noortje Dannenberg, Victor J. Carrion Bravo, Tom Weijers, Herman P. Spaink, Tom H. M. Ottenhoff, Ariane Briegel, Dennis Claessen
doi: https://doi.org/10.1101/2022.11.16.516772
Noortje Dannenberg
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands
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Victor J. Carrion Bravo
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands
2Departamento de Microbiología, Instituto de Hortofruticultura Subtropical y Mediterránea ‘La Mayora’, Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA-CSIC), Universidad de Málaga, Málaga, Spain
3Department of Microbial Ecology, Netherlands Institute of Ecology (NIOO-KNAW), Wageningen, the Netherlands
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Tom Weijers
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands
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Herman P. Spaink
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands
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Tom H. M. Ottenhoff
4Infectious Diseases, Leiden University Medical Center, Albinusdreef 2, 2333 ZA Leiden, The Netherlands
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Ariane Briegel
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands
5Netherlands Centre for Electron Nanoscopy (NeCEN), Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands
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Dennis Claessen
1Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands
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  • For correspondence: d.claessen@biology.leidenuniv.nl
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ABSTRACT

The cell wall is a unifying trait in bacteria and provides protection against environmental insults. Therefore, the wall is considered essential for most bacteria. Despite this critical role, many bacteria can transiently shed their cell wall and recent observations suggest a link of such wall-deficient cells to chronic infections. Whether shedding the cell wall also occurs in mycobacteria has not been established unambiguously. Here we provide compelling evidence that a wide range of mycobacterial species, including clinical and non-clinical isolates, form viable cell wall-deficient cells in response to environmental stressors. Using cryo-transmission electron micrography we show that the complex multi-layered wall is largely lost in such cells. Notably, we show that their formation in Mycobacterium marinum and BCG vaccine strains of Mycobacterium bovis is stimulated by exposure to cell wall-targeting antibiotics. Given that these wall-deficient mycobacteria are undetectable using conventional diagnostic methods, such cells have likely been overlooked in clinical settings. Altogether, these results indicate that mycobacteria can readily switch between a walled and wall-deficient lifestyle, which provides a plausible explanation for enabling persistence of infections caused by members of this genus.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted November 16, 2022.
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Mycobacteria form viable cell wall-deficient cells that are undetectable by conventional diagnostics
Noortje Dannenberg, Victor J. Carrion Bravo, Tom Weijers, Herman P. Spaink, Tom H. M. Ottenhoff, Ariane Briegel, Dennis Claessen
bioRxiv 2022.11.16.516772; doi: https://doi.org/10.1101/2022.11.16.516772
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Mycobacteria form viable cell wall-deficient cells that are undetectable by conventional diagnostics
Noortje Dannenberg, Victor J. Carrion Bravo, Tom Weijers, Herman P. Spaink, Tom H. M. Ottenhoff, Ariane Briegel, Dennis Claessen
bioRxiv 2022.11.16.516772; doi: https://doi.org/10.1101/2022.11.16.516772

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