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A recombinant Mycobacterium smegmatis induces potent bactericidal immunity against Mycobacterium tuberculosis

Nature Medicine volume 17, pages 12611268 (2011) | Download Citation

Abstract

We report the involvement of an evolutionarily conserved set of mycobacterial genes, the esx-3 region, in evasion of bacterial killing by innate immunity. Whereas high-dose intravenous infections of mice with the rapidly growing mycobacterial species Mycobacterium smegmatis bearing an intact esx-3 locus were rapidly lethal, infection with an M. smegmatis Δesx-3 mutant (here designated as the IKE strain) was controlled and cleared by a MyD88-dependent bactericidal immune response. Introduction of the orthologous Mycobacterium tuberculosis esx-3 genes into the IKE strain resulted in a strain, designated IKEPLUS, that remained susceptible to innate immune killing and was highly attenuated in mice but had a marked ability to stimulate bactericidal immunity against challenge with virulent M. tuberculosis. Analysis of these adaptive immune responses indicated that the highly protective bactericidal immunity elicited by IKEPLUS was dependent on CD4+ memory T cells and involved a distinct shift in the pattern of cytokine responses by CD4+ cells. Our results establish a role for the esx-3 locus in promoting mycobacterial virulence and also identify the IKE strain as a potentially powerful candidate vaccine vector for eliciting protective immunity to M. tuberculosis.

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Acknowledgements

We acknowledge the support of US National Institutes of Health (NIH) grants AI063537, AI093649, AI092448, AI26170 and AI051519 (Einstein Center for AIDS Research), and the Bill and Melinda Gates Foundation Collaboration for AIDS Vaccine Discovery. K.A.S. acknowledges support from the Albert Einstein College of Medicine's Institutional AIDS Training Grant T32-AI007501. We thank C. Harding, L. Ramachandra, G. Lauvau and S. Morris for advice and suggestions, as well as C. Colon-Berezin and S. Tiwari for helpful editing of the manuscript. Flow cytometry studies were carried out using core facilities supported by the Einstein Cancer Center (NIH/National Cancer Institute CA013330) and the Einstein Center for AIDS Research (NIH AI051519).

Author information

Author notes

    • Dee N Dao
    •  & Michael F Goldberg

    These authors contributed equally to this work.

Affiliations

  1. Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, New York, USA.

    • Kari A Sweeney
    • , Dee N Dao
    • , Tsungda Hsu
    • , Paras Jain
    • , Bing Chen
    • , Mei Chen
    • , John Kim
    • , Regy Lukose
    •  & William R Jacobs Jr
  2. Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, New York, USA.

    • Kari A Sweeney
    • , Dee N Dao
    • , Michael F Goldberg
    • , Tsungda Hsu
    • , Manjunatha M Venkataswamy
    • , Paras Jain
    • , Bing Chen
    • , Mei Chen
    • , John Kim
    • , Regy Lukose
    • , John Chan
    • , Steven A Porcelli
    •  & William R Jacobs Jr
  3. Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA.

    • Marcela Henao-Tamayo
    • , Diane Ordway
    •  & Ian M Orme
  4. Department of Pathology, Albert Einstein College of Medicine, Bronx, New York, USA.

    • Rani S Sellers
  5. Department of Medicine, Albert Einstein College of Medicine, Bronx, New York, USA.

    • John Chan
    • , Steven A Porcelli
    •  & William R Jacobs Jr

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Contributions

K.A.S. constructed bacterial strains, performed or contributed to the design of most experiments, and analyzed and interpreted data. D.N.D. carried out portions of the infection and challenge experiments. M.F.G. contributed to the T cell adoptive transfer studies and designed, performed and analyzed all flow cytometry analyses. T.H. participated in design and construction of bacterial strains and in the performance of mouse infection and challenge experiments. P.J. contributed to construction of bacterial strains. M.M.V. and M.H.-T. assisted with experiments analyzing responding T cell populations. R.S.S. analyzed and scored the histopathology samples. B.C., M.C., J.K. and R.L. carried out mouse infections, organ harvesting and quantification of bacilli in tissues. D.O., J.C., I.M.O., S.A.P. and W.R.J. Jr. designed and interpreted experiments. K.A.S., S.A.P. and W.R.J. Jr. wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to William R Jacobs Jr.

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DOI

https://doi.org/10.1038/nm.2420

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