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A replication clock for Mycobacterium tuberculosis

Nature Medicine volume 15, pages 211–214 (2009)Cite this article

Abstract

Few tools exist to assess replication of chronic pathogens during infection. This has been a considerable barrier to understanding latent tuberculosis, and efforts to develop new therapies generally assume that the bacteria are very slowly replicating or nonreplicating during latency1,2,3. To monitor Mycobacterium tuberculosis replication within hosts, we exploit an unstable plasmid that is lost at a steady, quantifiable rate from dividing cells in the absence of antibiotic selection. By applying a mathematical model, we calculate bacterial growth and death rates during infection of mice. We show that during chronic infection, the cumulative bacterial burden—enumerating total live, dead and removed organisms encountered by the mouse lung—is substantially higher than estimates from colony-forming units. Our data show that M. tuberculosis replicates throughout the course of chronic infection of mice and is restrained by the host immune system. This approach may also shed light on the replication dynamics of other chronic pathogens.

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Figure 1: In vitro stability of pBP10 in M. smegmatis and Mtb in the absence of antibiotic selection.
Figure 2: Stability of pBP10 in Mtb in the absence of bacterial replication.
Figure 3: Plasmid loss, bacterial replication and cumulative bacterial burden in mouse lungs.

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Acknowledgements

We would like to thank L. Ramakrishnan, J. Chang, K. Guinn, T. Walker and members of the Sherman lab for review of and contributions to this work, C. DeMille for expert help with manuscript preparation and K.G. Papavinasasundaram (University of Massachusetts Medical School) for providing pBP10. This research was supported by grants from the US National Institutes of Health and the Paul G. Allen Family Foundation.

Author information

Author notes

  1. Wendy P Gill

    Present address: Current address: Infectious Disease Consultants, 1601 East Nineteenth Avenue, Suite 3650, Denver, Colorado 80218, USA.,

  2. Wendy P Gill and Nada S Harik: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Allergy and Infectious Diseases, University of Washington Medical Center, 1959 Northeast Pacific Street, Box 356523, Seattle, 98195, Washington, USA

    Wendy P Gill

  2. Division of Pediatric Infectious Diseases, University of Arkansas for Medical Sciences, 800 Marshall Street, Slot 512-11, Little Rock, 72202, Arkansas, USA

    Nada S Harik

  3. Seattle Biomedical Research Institute, 307 Westlake Avenue North, Suite 500, Seattle, 98109, Washington, USA

    Molly R Whiddon, Reiling P Liao & David R Sherman

  4. Department of Global Health, Interdisciplinary Program of Pathobiology, University of Washington, Health Sciences Building H-660, Box 357660, Seattle, 98195, Washington, USA

    Molly R Whiddon & David R Sherman

  5. Department of Microbiology, University of Washington, Box 357242, Seattle, 98195, Washington, USA

    John E Mittler

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Contributions

W.P.G., N.S.H., M.R.W., R.P.L. and D.R.S. designed all experiments. W.P.G., N.S.H., R.P.L. and M.R.W. performed all experiments. W.P.G., N.S.H., M.R.W., J.E.M. and D.R.S. analyzed the data. J.E.M. derived the equations in consultation with W.P.G., and D.R.S. W.P.G. and D.R.S. wrote the paper, which was edited by all authors.

Corresponding author

Correspondence to David R Sherman.

Supplementary information

Supplementary Text and Figures

Supplementary Methods, Supplementary Fig.1 and Supplementary Equations 1 and 2 (PDF 364 kb)

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Gill, W., Harik, N., Whiddon, M. et al. A replication clock for Mycobacterium tuberculosis. Nat Med 15, 211–214 (2009). https://doi.org/10.1038/nm.1915

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