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Suppression of autophagy and antigen presentation by Mycobacterium tuberculosis PE_PGRS47

Nature Microbiology volume 1, Article number: 16133 (2016) Cite this article

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Abstract

Suppression of major histocompatibility complex (MHC) class II antigen presentation is believed to be among the major mechanisms used by Mycobacterium tuberculosis to escape protective host immune responses. Through a genome-wide screen for the genetic loci of M. tuberculosis that inhibit MHC class II-restricted antigen presentation by mycobacteria-infected dendritic cells, we identified the PE_PGRS47 protein as one of the responsible factors. Targeted disruption of the PE_PGRS47 (Rv2741) gene led to attenuated growth of M. tuberculosis in vitro and in vivo, and a PE_PGRS47 mutant showed enhanced MHC class II-restricted antigen presentation during in vivo infection of mice. Analysis of the effects of deletion or over-expression of PE_PGRS47 implicated this protein in the inhibition of autophagy in infected host phagocytes. Our findings identify PE_PGRS47 as a functionally relevant, non-redundant bacterial factor in the modulation of innate and adaptive immunity by M. tuberculosis, suggesting strategies for improving antigen presentation and the generation of protective immunity during vaccination or infection.

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Figure 1: Identification of Mtb cosmid clones that inhibit MHC class II antigen presentation.
Figure 2: Inhibition of autophagy by PE_PGRS47.
Figure 3: Increased acidification and lysosomal fusion of phagosomes containing Mtb ΔPE_PGRS47.
Figure 4: PE_PGRS47 is required for full virulence at late stages of Mtb infection.
Figure 5: Autophagy-dependent enhancement of MHC class II antigen presentation during in vitro infection by the Mtb ΔPE_PGRS47 mutant.
Figure 6: Suppression of CD4+ T-cell responses by PE_PGRS47.

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Acknowledgements

The authors thank the staff of the Flow Cytometry Core Facility of the Albert Einstein College of Medicine (supported by the Einstein Cancer Center grant NIH/NCI CA13330). The authors acknowledge the NIH Tetramer Core Facility (contract no. HHSN272201300006C) for provision of the I-Ab/TB9.8 tetramers. This work was supported by NIH grants AI093649 to S.A.P. and AI063537 to S.A.P., W.R.J. and J.C. The authors thank N. Mizushima, Tokyo Medical and Dental University, for providing the GFP-LC3 lentivirus construct and A.Y. Rudensky for providing the Y-Ae hybridoma. The authors also thank P. Jain and P. A. Gonzalez for their suggestions on the construction of the ΔPE_PGRS47 mutant strain and R. Sellers for assistance in analysis of histopathology. A.B. acknowledges support from ‘Estrategia de Sostenibilidad Universidad de Antioquia’. The authors thank R. Prados-Rosales for assistance with EM studies and E. Bejarano for advice on autophagy analysis.

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Author notes

  1. Andres Baena, Manjunatha M. Venkataswamy & Leandro J. Carreño

    Present address: ‡Present addresses: Grupo de Inmunología Celular e Inmunogenética (GICIG), Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad de Antioquia, Calle 70 No. 52-21, Medellín, Colombia (A.B.); Department of Neurovirology, NIMHANS, 560029 Bangalore, India (M.M.V.); Millenium Institute on Immunology and Immunotherapy, Programa Disciplinario de Inmunologia, Universidad de Chile, Chile (L.J.C.).,

  2. Neeraj K. Saini and Andres Baena: †These authors contributed equally to this work

Authors and Affiliations

  1. Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, 10461, New York, USA

    Neeraj K. Saini, Andres Baena, Tony W. Ng, Manjunatha M. Venkataswamy, Steven C. Kennedy, Shajo Kunnath-Velayudhan, Leandro J. Carreño, Michelle H. Larsen, William R. Jacobs Jr & Steven A. Porcelli

  2. Department of Medicine, Albert Einstein College of Medicine, Bronx, 10461, New York, USA

    Jiayong Xu, John Chan & Steven A. Porcelli

  3. Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, 10461, New York, USA

    Michelle H. Larsen & William R. Jacobs Jr

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Contributions

N.K.S., A.B. and T.W.N. designed and carried out experiments. S.A.P. supervised the design and execution of all experiments. M.M.V., S.C.K., S.K.-V., L.J.C. and J.X. assisted in the execution of selected experiments. J.C., M.H.L. and W.R.J. provided input on the design of experiments and data interpretation. All authors contributed to writing and editing the manuscript.

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Correspondence to Steven A. Porcelli.

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Saini, N., Baena, A., Ng, T. et al. Suppression of autophagy and antigen presentation by Mycobacterium tuberculosis PE_PGRS47. Nat Microbiol 1, 16133 (2016). https://doi.org/10.1038/nmicrobiol.2016.133

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