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A bacterial cyclic dinucleotide activates the cytosolic surveillance pathway and mediates innate resistance to tuberculosis

Nature Medicine volume 21, pages 401–406 (2015)Cite this article

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Abstract

Detection of cyclic-di-adenosine monophosphate (c-di-AMP), a bacterial second messenger, by the host cytoplasmic surveillance pathway (CSP) is known to elicit type I interferon (IFN) responses, which are crucial to antimicrobial defense1,2,3. However, the mechanisms and role of c-di-AMP signaling in Mycobacterium tuberculosis virulence remain unclear. Here we show that resistance to tuberculosis requires CSP-mediated detection of c-di-AMP produced by M. tuberculosis and that levels of c-di-AMP modulate the fate of infection. We found that a di-adenylate cyclase (disA or dacA)4-overexpressing M. tuberculosis strain that secretes excess c-di-AMP activates the interferon regulatory factor (IRF) pathway with enhanced levels of IFN-β, elicits increased macrophage autophagy, and exhibits substantial virulence attenuation in mice. We show that c-di-AMP-mediated IFN-β induction during M. tuberculosis infection requires stimulator of interferon genes (STING)5-signaling. We observed that c-di-AMP induction of IFN-β is independent of the cytosolic nucleic acid receptor cyclic GMP-AMP (cGAMP) synthase (cGAS)6,7, but cGAS nevertheless contributes substantially to the overall IFN-β response to M. tuberculosis infection. In sum, our results reveal c-di-AMP to be a key mycobacterial pathogen-associated molecular pattern (PAMP) driving host type I IFN responses and autophagy. These findings suggest that modulating the levels of this small molecule may lead to novel immunotherapeutic strategies against tuberculosis.

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Figure 1: Modulation of host cytokine responses and intracellular growth of M. tuberculosis by c-di-AMP.
Figure 2: c-di-AMP produced by M. tuberculosis induces autophagy in macrophage cells.
Figure 3: Attenuation of virulence and pathogenicity in a c-di-AMP–overproducing M. tuberculosis strain.
Figure 4: Contribution of STING and cytosolic DNA receptor cGAS to c-di-AMP mediated activation of IFN-β during M. tuberculosis infection.

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Acknowledgements

The support of grants AI037856, AI097138 and AI036973 from the US National Institute of Allergy and Infectious Diseases and the Howard Hughes Medical Institute to W.R.B. is gratefully acknowledged. We thank MicFac facility members at Johns Hopkins Medical Institutions (JHMI) for helping with the fluorescence and confocal microscopy. We thank H.W. Virgin IV and D. MacDuff (Washington University School of Medicine in St. Louis), for generously providing bone marrow cells from cGAS-KO mice. We thank L.H. Moulton (Bloomberg School of Public Health, Johns Hopkins University) for generous assistance with the statistical analysis. We thank A. Tyagi (University of Delhi South Campus, New Delhi) and W. Jacobs, Jr. (Albert Einstein College of Medicine, New York) for mycobacterial cloning vectors.

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Authors and Affiliations

  1. Center for Tuberculosis Research, Johns Hopkins University, Baltimore, Maryland, USA

    Bappaditya Dey, Ruchi Jain Dey, Laurene S Cheung, Supriya Pokkali, Haidan Guo, Jong-Hee Lee & William R Bishai

  2. Howard Hughes Medical Institute, Chevy Chase, Maryland, USA

    Bappaditya Dey & William R Bishai

  3. Department of Pediatrics, Johns Hopkins University, Baltimore, Maryland, USA

    Supriya Pokkali

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Contributions

B.D., R.J.D. and W.R.B. designed the research. B.D., R.J.D. and L.S.C. performed the experiments. H.G. contributed to mouse experiments. S.P. contributed to mouse, BMDM and DMDC experiments. J.-H.L. contributed to LC-MS analysis. B.D., R.J.D. and W.R.B. analyzed the data and wrote the paper. W.R.B. provided overall supervision of the study.

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Correspondence to William R Bishai.

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Dey, B., Dey, R., Cheung, L. et al. A bacterial cyclic dinucleotide activates the cytosolic surveillance pathway and mediates innate resistance to tuberculosis. Nat Med 21, 401–406 (2015). https://doi.org/10.1038/nm.3813

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