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Autophagy favors Brucella melitensis survival in infected macrophages

Cellular & Molecular Biology Letters volume 17, pages 249–257 (2012)Cite this article

Abstract

This study investigated the role of autophagy in the survival of the invasive bacterium Brucella melitensis strain 16M in murine macrophages. Here, Brucella melitensis 16M was found to trigger autophagosome formation, enhance autophagy flux and increase the expression level of the autophagy marker protein LC3-II. When autophagy was pharmacologically inhibited by 3-methyladenine (3-MA), Brucella replication efficiency was significantly decreased (p < 0.05). These results suggest that autophagy favors Brucella melitensis 16M survival in murine macrophages.

Abbreviations

3-MA:

3-methyladenine

BCV:

Brucella-containing vacuole

CFU:

colony forming units

DMEM:

Dulbecco’s Modified Eagle’s Medium

ER:

endoplasmic reticulum

FBS:

fetal bovine serum

GFP:

green fluorescent protein

MOI:

multiplicity of infection

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

PBS:

phosphate buffered saline

TBST:

Tris buffered saline with Tween

References

  1. Capasso, L. Bacteria in two-millennia-old cheese, and related epizoonoses in Rome populations. J. Infect. 45 (2002) 122–127.

    Article  PubMed  CAS  Google Scholar 

  2. Aroian, R. and Goot, F.G. Pore-forming toxins and cellular non-immune defenses (CNIDs). Curr. Opin. Microbiol. 10 (2007) 57–61.

    Article  PubMed  CAS  Google Scholar 

  3. Sarinas, P.S. and Chitkara, R.K. Brucellosis. Semin. Respir. Infect. 18 (2003) 168–182.

    PubMed  Google Scholar 

  4. Gibbs, E.P. Emerging zoonotic epidemics in the interconnected global community. Vet. Rec. 157 (2005) 673–679.

    Article  PubMed  CAS  Google Scholar 

  5. Otto, G.P., Wu, M.Y., Clarke, M., Lu, H., Anderson, O.R., Hilbi, H., Shuman, H.A. and Kessin, R.H. Macroautophagy is dispensable for intracellular replication of Legionella pneumophila in Dictyostelium discoideum. Mol. Microbiol. 51 (2004) 63–71.

    Article  PubMed  CAS  Google Scholar 

  6. Romano, P.S., Gutierrez, M.G., Beron, W., Rabinovitch, M. and Colombo, M.I. The autophagic pathway is actively modulated by phase II Coxiella burnetii to efficiently replicate in the host cell. Cell. Microbiol. 9 (2007) 891–897.

    Article  PubMed  CAS  Google Scholar 

  7. Gutierrez, M.G., Master, S.S., Singh, S.B., Taylor, G.A., Colombo, M.I. and Deretic, V. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages. Cell 119 (2004) 753–765.

    Article  PubMed  CAS  Google Scholar 

  8. Py, B.F., Lipinski, M.M. and Yuan, J. Autophagy limits Listeria monocytogenes intracellular growth in the early phase of primary infection. Autophagy 3 (2007) 117–126.

    PubMed  CAS  Google Scholar 

  9. Jones, S.M. and Winter, A.J. Survival of virulent and attenuated strains of Brucella abortus in normal and gamma interferon-activated murine peritoneal macrophages. Infect. Immun. 60 (1992) 3011–3014.

    PubMed  CAS  Google Scholar 

  10. Smith, R., Adams, L.G., Sowa, B.A. and Ficht, T.A. Induction of lymphocyte responsiveness by the outer membrane-peptidoglycan complex of rough strains of Brucella abortus. Vet. Immunol. Immunopathol. 26 (1990) 31–48.

    Article  PubMed  Google Scholar 

  11. Gorvel, J.P. and Moreno, E. Brucella intracellular life: from invasion to intracellular replication. Vet. Microbiol. 90 (2002) 281–297.

    Article  PubMed  CAS  Google Scholar 

  12. Starr, T., Ng, T.W., Wehrly, T.D., Knodler, L.A. and Celli, J. Brucella intracellular replication requires trafficking through the late endosomal/lysosomal compartment. Traffic 9 (2008) 678–694.

    Article  PubMed  CAS  Google Scholar 

  13. Celli, J., de Chastellier, C., Franchini, D.M., Javier P.C., Moreno, E. and Gorvel, J.P. Brucella evades macrophage killing via VirB-dependent sustained interactions with the endoplasmic reticulum. J. Exp. Med. 198 (2003) 545–556.

    Article  PubMed  CAS  Google Scholar 

  14. Pizarro-Cerda, J., Meresse, S., Parton, R.G., van der, G.G., Sola-Landa, A., Lopez-Goni, I., Moreno, E. and Orvel, J.P. Brucella abortus transits through the autophagic pathway and replicates in the endoplasmic reticulum of nonprofessional phagocytes. Infect. Immun. 66 (1998) 5711–5724.

    PubMed  CAS  Google Scholar 

  15. Pizarro-Cerda, J. and Cossart, P. Bacterial adhesion and entry into host cells. Cell. 124 (2006) 715–727.

    Article  PubMed  CAS  Google Scholar 

  16. Li, M., Jiang, X., Liu, D., Na, Y., Gao, G.F. and Xi, Z. Autophagy protects LNCaP cells under androgen deprivation conditions. Autophagy 4 (2008) 54–60.

    PubMed  CAS  Google Scholar 

  17. Yuk, J.M., Shin, D.M., Lee, H.M., Yang, C.S. and Jin, H.S. Vitamin D3 induces autophagy in human monocytes/macrophages via cathelicidin. Cell Host Microbe 6 (2009) 231–243.

    Article  PubMed  CAS  Google Scholar 

  18. Espert, L., Denizot, M., Grimaldi, M., Robert, H.V., Gay, B. and Varbanov, M. Autophagy is involved in T cell death after binding of HIV-1 envelope proteins to CXCR4. J. Clin. Invest. 116 (2006) 2161–2172.

    Article  PubMed  CAS  Google Scholar 

  19. Mizushima, N. Methods for monitoring autophagy. Int. J. Biochem. Cell Biol. 36 (2004) 2491–2502.

    Article  PubMed  CAS  Google Scholar 

  20. Kabeya, Y., Mizushima, N., Ueno, T., Yamamoto, A. and Kirisako, T. LC3 a mammalian homologue of yeast Apg8p is localized in autophagosome membranes after processing. EMBO J. 19 (2000) 5720–5728.

    Article  PubMed  CAS  Google Scholar 

  21. Gutierrez, M.G., Master, S.S., Singh, S.B., Taylor, G.A., Colombo, M.I. and Deretic, V. Autophagy is a defense mechanism inhibiting BCG and Mycobacterium tuberculosis survival in infected macrophages. Cell 119 (2004) 753–766.

    Article  PubMed  CAS  Google Scholar 

  22. Ogawa, M., Yoshimori, T., Suzuki, T., Sagara, H., Mizushima, N. and Sasakawa, C. Escape of intracellular Shigella from autophagy. Science 307 (2005) 727–737.

    Article  PubMed  CAS  Google Scholar 

  23. Horwitz, M.A. The Legionnaires’ disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes. J. Exp. Med. 158 (1983) 2108–2126.

    Article  PubMed  CAS  Google Scholar 

  24. Swanson, M.S. and Isberg, R.R. Association of Legionella pneumophila with the macrophage endoplasmic reticulum. Infect. Immun. 63 (1995) 3609–3620.

    PubMed  CAS  Google Scholar 

  25. Zhu, C., Wang, X., Xu, F., Bahr, B.A., Shibata, M., Uchiyama, Y., Hagberg, H. and Blomgren, K. The influence of age on apoptotic and other mechanisms of cell death after cerebral hypoxia-ischemia. Cell Death Diff. 12 (2005) 162–176.

    Article  CAS  Google Scholar 

  26. Arenas, G.N., Staskevich, A.S., Aballay, A. and Mayorga, L.S. Intracellular trafficking of Brucella abortus in J774 macrophages. Infect. Immun. 68 (2000) 4255–4263.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. College of Animal Science and Technology, Shihezi University, Shihezi, 832003, China

    Fei Guo, Chuangfu Chen, Jun Qiao, Ke Zhang, Yong Wang & Guoqing Du

  2. Key Laboratory of Xijiang Endemic and Ethnic Disease, Shihezi University, Shihezi, 832003, China

    Hui Zhang & Shengwei Hu

  3. College of Medicine, Shihezi University, Shihezi, 832003, China

    Yuanzhi Wang & Yan Ren

Authors
  1. Fei Guo
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  2. Hui Zhang
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  3. Chuangfu Chen
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  4. Shengwei Hu
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  5. Yuanzhi Wang
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  6. Jun Qiao
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  7. Yan Ren
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  8. Ke Zhang
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  9. Yong Wang
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  10. Guoqing Du
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Correspondence to Chuangfu Chen.

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Both authors contributed equally to the work

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Guo, F., Zhang, H., Chen, C. et al. Autophagy favors Brucella melitensis survival in infected macrophages. Cell Mol Biol Lett 17, 249–257 (2012). https://doi.org/10.2478/s11658-012-0009-4

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  • DOI: https://doi.org/10.2478/s11658-012-0009-4

Key words

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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