Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Autophagy enhances the efficacy of BCG vaccine by increasing peptide presentation in mouse dendritic cells

Nature Medicine volume 15, pages 267–276 (2009)Cite this article

A Corrigendum to this article was published on 01 July 2010

This article has been updated

Abstract

The variable efficacy of Bacille Calmette Guerin (BCG) vaccination against tuberculosis has prompted efforts to improve the vaccine. In this study, we used autophagy to enhance vaccine efficacy against tuberculosis in a mouse model. We examined the effect of autophagy on the processing of the immunodominant mycobacterial antigen Ag85B by antigen presenting cells (APCs), macrophages and dendritic cells (DCs). We found that rapamycin-induced autophagy enhanced Ag85B presentation by APCs infected with wild-type Mycobacterium tuberculosis H37Rv, H37Rv-derived ΔfbpA attenuated candidate vaccine or BCG. Furthermore, rapamycin enhanced localization of mycobacteria with autophagosomes and lysosomes. Rapamycin-enhanced antigen presentation was attenuated when autophagy was suppressed by 3-methyladenine or by small interfering RNA against beclin-1. Notably, mice immunized with rapamycin-treated DCs infected with either ΔfbpA or BCG showed enhanced T helper type 1–mediated protection when challenged with virulent Mycobacterium tuberculosis. Finally, overexpression of Ag85B in BCG induced autophagy in APCs and enhanced immunogenicity in mice, suggesting that vaccine efficacy can be enhanced by augmenting autophagy-mediated antigen presentation.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Rapamycin enhances processing and presentation of mycobacterial antigen 85B (Ag85B) in macrophages.
Figure 2: Rapamycin enhances colocalization of mycobacteria with autophagosomes.
Figure 3: Rapamycin-induced autophagy leads to colocalization of mycobacteria with lysosomes and enrichment of antigen-processing components.
Figure 4: Rapamycin enhances the ability of dendritic cells to prime T cells for Ag85B.
Figure 5: Pretreatment of mycobacteria-infected DCs with rapamycin enhances TH1 responses in mice and increases vaccine efficacy.
Figure 6: BCG vaccine overexpressing antigen 85B enhances in vivo protection against Mtb when compared to wild-type BCG.

Similar content being viewed by others

Change history

  • 07 July 2010

     In the version of this article initially published online, Figure 1b was a copy of Figure 4b, and Figure 3d did not show all the correct bands. Both figures have been corrected for the PDF and HTML versions of this article. The legends and interpretation of the data remain the same.

References

  1. Flynn, J.L. & Chan, J. Immunology of tuberculosis. Annu. Rev. Immunol. 19, 93–129 (2001).

    Article  CAS  Google Scholar 

  2. Nathan, C. & Shiloh, M.U. Reactive oxygen and nitrogen intermediates in the relationship between mammalian hosts and microbial pathogens. Proc. Natl. Acad. Sci. USA 97, 8841–8848 (2000).

    Article  CAS  Google Scholar 

  3. Miller, B.H. et al. Mycobacteria inhibit nitric oxide synthase recruitment to phagosomes during macrophage infection. Infect. Immun. 72, 2872–2878 (2004).

    Article  CAS  Google Scholar 

  4. Vergne, I., Chua, J. & Deretic, V. Mycobacterium tuberculosis phagosome maturation arrest: selective targeting of PI3P-dependent membrane trafficking. Traffic 4, 600–606 (2003).

    Article  CAS  Google Scholar 

  5. Rohde, K., Yates, R.M., Purdy, G.E. & Russell, D.G. Mycobacterium tuberculosis and the environment within the phagosome. Immunol. Rev. 219, 37–54 (2007).

    Article  CAS  Google Scholar 

  6. Dean, R.T. Lysosomes and protein degradation. Acta Biol. Med. Ger. 36, 1815–1820 (1977).

    CAS  PubMed  Google Scholar 

  7. Hmama, Z., Gabathuler, R., Jefferies, W.A., de Jong, G. & Reiner, N.E. Attenuation of HLA-DR expression by mononuclear phagocytes infected with Mycobacterium tuberculosis is related to intracellular sequestration of immature class II heterodimers. J. Immunol. 161, 4882–4893 (1998).

    CAS  PubMed  Google Scholar 

  8. Noss, E.H., Harding, C.V. & Boom, W.H. Mycobacterium tuberculosis inhibits MHC class II antigen processing in murine bone marrow macrophages. Cell. Immunol. 201, 63–74 (2000).

    Article  CAS  Google Scholar 

  9. Noss, E.H. et al. Toll-like receptor 2–dependent inhibition of macrophage class II MHC expression and antigen processing by 19-kDa lipoprotein of Mycobacterium tuberculosis. J. Immunol. 167, 910–918 (2001).

    Article  CAS  Google Scholar 

  10. Pancholi, P., Mirza, A., Bhardwaj, N. & Steinman, R.M. Sequestration from immune CD4+ T cells of mycobacteria growing in human macrophages. Science 260, 984–986 (1993).

    Article  CAS  Google Scholar 

  11. Harth, G., Lee, B.Y., Wang, J., Clemens, D.L. & Horwitz, M.A. Novel insights into the genetics, biochemistry, and immunocytochemistry of the 30-kilodalton major extracellular protein of Mycobacterium tuberculosis. Infect. Immun. 64, 3038–3047 (1996).

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Singh, C.R. et al. Processing and presentation of a mycobacterial antigen 85B epitope by murine macrophages is dependent on the phagosomal acquisition of vacuolar proton ATPase and in situ activation of cathepsin D. J. Immunol. 177, 3250–3259 (2006).

    Article  CAS  Google Scholar 

  13. Soualhine, H. et al. Mycobacterium bovis bacillus calmette-guerin secreting active cathepsin s stimulates expression of mature MHC class II molecules and antigen presentation in human macrophages. J. Immunol. 179, 5137–5145 (2007).

    Article  CAS  Google Scholar 

  14. Katti, M.K. et al. The ΔfbpA mutant derived from Mycobacterium tuberculosis H37Rv has an enhanced susceptibility to intracellular antimicrobial oxidative mechanisms, undergoes limited phagosome maturation and activates macrophages and dendritic cells. Cell. Microbiol. 10, 1286–1303 (2008).

    Article  CAS  Google Scholar 

  15. Schmid, D. & Munz, C. Innate and adaptive immunity through autophagy. Immunity 27, 11–21 (2007).

    Article  CAS  Google Scholar 

  16. Schmid, D. & Munz, C. Immune surveillance via self digestion. Autophagy 3, 133–135 (2007).

    Article  CAS  Google Scholar 

  17. Schmid, D., Pypaert, M. & Munz, C. Antigen-loading compartments for major histocompatibility complex class II molecules continuously receive input from autophagosomes. Immunity 26, 79–92 (2007).

    Article  CAS  Google Scholar 

  18. Gutierrez, M.G. et al. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages. Cell 119, 753–766 (2004).

    Article  CAS  Google Scholar 

  19. Amano, A., Nakagawa, I. & Yoshimori, T. Autophagy in innate immunity against intracellular bacteria. J. Biochem. 140, 161–166 (2006).

    Article  CAS  Google Scholar 

  20. Xu, Y. et al. Toll-like receptor 4 is a sensor for autophagy associated with innate immunity. Immunity 27, 135–144 (2007).

    Article  CAS  Google Scholar 

  21. Levine, B. & Deretic, V. Unveiling the roles of autophagy in innate and adaptive immunity. Nat. Rev. Immunol. 7, 767–777 (2007).

    Article  CAS  Google Scholar 

  22. Skeiky, Y.A. & Sadoff, J.C. Advances in tuberculosis vaccine strategies. Nat. Rev. Microbiol. 4, 469–476 (2006).

    Article  CAS  Google Scholar 

  23. Ramachandra, L., Noss, E., Boom, W.H. & Harding, C.V. Processing of Mycobacterium tuberculosis antigen 85B involves intraphagosomal formation of peptide–major histocompatibility complex II complexes and is inhibited by live bacilli that decrease phagosome maturation. J. Exp. Med. 194, 1421–1432 (2001).

    Article  CAS  Google Scholar 

  24. Copenhaver, R.H. et al. A mutant of Mycobacterium tuberculosis H37Rv that lacks expression of antigen 85A is attenuated in mice but retains vaccinogenic potential. Infect. Immun. 72, 7084–7095 (2004).

    Article  CAS  Google Scholar 

  25. Armitige, L.Y., Jagannath, C., Wanger, A.R. & Norris, S.J. Disruption of the genes encoding antigen 85A and antigen 85B of Mycobacterium tuberculosis H37Rv: effect on growth in culture and in macrophages. Infect. Immun. 68, 767–778 (2000).

    Article  CAS  Google Scholar 

  26. Tanida, I., Minematsu-Ikeguchi, N., Ueno, T. & Kominami, E. Lysosomal turnover, but not a cellular level, of endogenous LC3 is a marker for autophagy. Autophagy 1, 84–91 (2005).

    Article  CAS  Google Scholar 

  27. Ullrich, H.J., Beatty, W.L. & Russell, D.G. Interaction of Mycobacterium avium–containing phagosomes with the antigen presentation pathway. J. Immunol. 165, 6073–6080 (2000).

    Article  CAS  Google Scholar 

  28. Horwitz, M.A., Harth, G., Dillon, B.J. & Maslesa-Galic, S. A novel live recombinant mycobacterial vaccine against bovine tuberculosis more potent than BCG. Vaccine 24, 1593–1600 (2006).

    Article  CAS  Google Scholar 

  29. Moulton, R.A. et al. Complement C5a anaphylatoxin is an innate determinant of dendritic cell–induced TH1 immunity to Mycobacterium bovis BCG infection in mice. J. Leukoc. Biol. 82, 956–967 (2007).

    Article  CAS  Google Scholar 

  30. Gupta, U.D., Katoch, V.M. & McMurray, D.N. Current status of TB vaccines. Vaccine 25, 3742–3751 (2007).

    Article  CAS  Google Scholar 

  31. Levine, T.P. & Chain, B.M. The cell biology of antigen processing. Crit. Rev. Biochem. Mol. Biol. 26, 439–473 (1991).

    Article  CAS  Google Scholar 

  32. Medd, P.G. & Chain, B.M. Protein degradation in MHC class II antigen presentation: opportunities for immunomodulation. Semin. Cell Dev. Biol. 11, 203–210 (2000).

    Article  CAS  Google Scholar 

  33. Mazzaccaro, R.J. et al. Cytotoxic T lymphocytes in resistance to tuberculosis. Adv. Exp. Med. Biol. 452, 85–101 (1998).

    Article  CAS  Google Scholar 

  34. Dhandayuthapani, S. et al. Green fluorescent protein as a marker for gene expression and cell biology of mycobacterial interactions with macrophages. Mol. Microbiol. 17, 901–912 (1995).

    Article  CAS  Google Scholar 

  35. Am, A.O., Byrne, B.G. & Swanson, M.S. Macrophages rapidly transfer pathogens from lipid raft vacuoles to autophagosomes. Autophagy 1, 53–58 (2005).

    Article  Google Scholar 

  36. Griffin, J.P. & Orme, I.M. Evolution of CD4 T cell subsets following infection of naive and memory immune mice with Mycobacterium tuberculosis. Infect. Immun. 62, 1683–1690 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This study was supported by US National Institutes of Health National Institute of Allergy and Infectious Diseases grant AI49534 (C.J.) and National Heart, Lung, and Blood Institute grant HL080205 (N.T.E.). We are grateful to L.Y. Armitige (University of Texas Health Sciences Center–Houston) for the fbpA and fbpB mutant strains, K. Rock (University of Massachusetts–Worcester) for the BMA.A3 cell line and C. Harding and H. Boom (Case Western Reserve University) for BB7 T cells. Recombinant ESAT-6 was kindly provided by A. Arora (Central Drug Research Institute, India), and TD17 mAb was a gift from K. Huygen (Pasteur Institute). Mycobacterial vector pMV206 was kindly provided by W. Jacobs (Albert Einstein Medical College).

Author information

Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, University of Texas Health Sciences Center (UTHSC), 6431, Fannin, Houston, 77030, Texas, USA

    Chinnaswamy Jagannath, Devin R Lindsey & Robert L Hunter Jr

  2. Regional Academic Health Center and Department of Microbiology and Immunology, UTHSC-San Antonio, Edinburg, 78541, Texas, USA

    Subramanian Dhandayuthapani

  3. Department of Medicine, Baylor College of Medicine, 1 Baylor Plaza, Houston, 77030, Texas, USA

    Yi Xu & N Tony Eissa

Authors
  1. Chinnaswamy Jagannath
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Devin R Lindsey
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Subramanian Dhandayuthapani
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Yi Xu
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Robert L Hunter Jr
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. N Tony Eissa
    View author publications

    You can also search for this author inPubMed Google Scholar

Contributions

C.J. designed and performed the experiments with the technical assistance of D.R.L. S.D. prepared the overexpression strains of M. smegmatis and BCG. X.Y. and N.T.E. designed and produced RFP-LC3. C.J. and N.T.E. interpreted the data and wrote the manuscript. R.L.H. edited the manuscript.

Corresponding author

Correspondence to Chinnaswamy Jagannath.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–7 (PDF 847 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Jagannath, C., Lindsey, D., Dhandayuthapani, S. et al. Autophagy enhances the efficacy of BCG vaccine by increasing peptide presentation in mouse dendritic cells. Nat Med 15, 267–276 (2009). https://doi.org/10.1038/nm.1928

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.1928

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing