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Deletion of the Type IV Secretion System Effector VceA Promotes Autophagy and Inhibits Apoptosis in Brucella-Infected Human Trophoblast Cells

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Abstract

Brucellosis is the most common zoonotic disease that caused by intracellular parasitic bacteria Brucella. The survival and replication of Brucella in the host depend on the type IV secretion system (T4SS). The T4SS system of Brucella has many components and secreted proteins. But the mechanism helped Brucella to evade the host defense is still not clear. The objective of the present study was to investigate the effects of VceA on autophagy and apoptosis in Brucella-infected embryonic trophoblast cells. We constructed the VceA mutant strain (2308ΔVceA) and complementary strain (2308ΔVceA-C) of Brucella abortus 2308 (S2308). The human trophoblast cells (HPT-8 cells) and mice were infected by S2308, 2308ΔVceA and 2308ΔVceA-C. The cell autophagy and apoptosis were detected. The Atg5, LC3-II and Bcl-2 mRNA expression were significantly increased in 2308ΔVceA group than the S2308 group, and mRNA expression of P62 and Caspase-3 were significantly decreased than the S2308 group. Western blotting, qPCR and flow cytometry analysis showed that 2308ΔVceA promoted autophagy and inhibited apoptosis. Mouse immunohistochemistry experiments showed that P62 protein was scattered coloring and Cytochrome C protein was scarcely in 2308ΔVceA group at the myometrium. These results indicated that 2308ΔVceA promoted autophagy and inhibited apoptosis in HPT-8 cells during Brucella infection.

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References

  1. Arellano-Reynoso B, Lapaque N, Salcedo S, Briones G, Ciocchini A, Ugalde R, Moreno E, Moriyón I, Gorvel J (2005) Cyclic beta-1,2-glucan is a Brucella virulence factor required for intracellular survival. Nat Immunol 6:618–625

    Article  CAS  PubMed  Google Scholar 

  2. Backert S, Meyer T (2006) Type IV secretion systems and their effectors in bacterial pathogenesis. Curr Opin Microbiol 9:207–217

    Article  CAS  PubMed  Google Scholar 

  3. Barrionuevo P, Delpino MV, Pozner RG, Velasquez LN, Cassataro J, Giambartolomei GH (2013) Brucella abortus induces intracellular retention of MHC-I molecules in human macrophages down-modulating cytotoxic CD8(+) T cell responses. Cell Microbiol 15:487–502

    Article  CAS  PubMed  Google Scholar 

  4. de Jong MF, Starr T, Winter MG, den Hartigh AB, Child R, Knodler LA, van Dijl JM, Celli J, Tsolis RM (2013) Sensing of bacterial type IV secretion via the unfolded protein response. MBio 4:e00418–e00412

    PubMed  PubMed Central  Google Scholar 

  5. De Jong MF, Sun YH, Den Hartigh AB, Van Dijl JM, Tsolis RM (2008) Identification of VceA and VceC, two members of the VjbR regulon that are translocated into macrophages by the Brucella type IV secretion system. Mol Microbiol 70:1378–1396

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. de Jong MF, Tsolis RM (2012) Brucellosis and type IV secretion. Future Microbiol 7:47–58

    Article  PubMed  Google Scholar 

  7. Dong H, Liu W, Peng X, Jing Z, Wu Q (2013) The effects of mucr on expression of type IV secretion system, quorum sensing system and stress responses in Brucella melitensis. Vet Microbiol 166:535–542

    Article  CAS  PubMed  Google Scholar 

  8. Franco IS, Shuman HA, Charpentier X (2009) The perplexing functions and surprising origins of legionella pneumophila type IV secretion effectors. Cell Microbiol 11:1435–1443

    Article  CAS  PubMed  Google Scholar 

  9. Frenchick PJ, Markham RJ, Cochrane AH (1985) Inhibition of phagosome-lysosome fusion in macrophages by soluble extracts of virulent Brucella abortus. Am J Vet Res 46:332–335

    CAS  PubMed  Google Scholar 

  10. Hale AN, Ledbetter DJ, Gawriluk TR, Rucker EB 3rd (2013) Autophagy: regulation and role in development. Autophagy 9:951–972

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. He Y, Reichow S, Ramamoorthy S, Ding X, Lathigra R, Craig JC, Sobral BW, Schurig GG, Sriranganathan N, Boyle SM (2006) Brucella melitensis triggers time-dependent modulation of apoptosis and down-regulation of mitochondrion-associated gene expression in mouse macrophages. Infect Immun 74:5035–5046

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Ke Y, Wang Y, Li W, Chen Z (2015) Type IV secretion system of Brucella spp. and its effectors. Front Cell Infect Microbiol 5:72

    Article  PubMed  PubMed Central  Google Scholar 

  13. Kremer L, Estaquier J, Brandt E, Ameisen JC, Locht C (1997) Mycobacterium bovis bacillus calmette guerin infection prevents apoptosis of resting human monocytes. Eur J Immunol 27:2450–2456

    Article  CAS  PubMed  Google Scholar 

  14. Li T, Xu Y, Liu L, Huang M, Wang Z, Tong Z, Zhang H, Guo F, Chen C (2016) Brucella melitensis 16 m regulates the effect of air domain on inflammatory factors, autophagy, and apoptosis in mouse macrophage through the ros signaling pathway. PLoS ONE 11:e0167486

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Li Z, Shi J, Fu W, Zhang Y, Zhang J, Wang Z, Li T, Chen C, Guo F, Zhang H (2015) A Brucella melitensis m5-90 wboa deletion strain is attenuated and enhances vaccine efficacy. Mol Immunol 66:276–283

    Article  CAS  PubMed  Google Scholar 

  16. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative pcr and the 2(−δδct) method. Methods 25:402–408

    Article  CAS  PubMed  Google Scholar 

  17. Ma QL, Liu AC, Ma XJ, Wang YB, Hou YT, Wang ZH (2015) Brucella outer membrane protein omp25 induces microglial cells in vitro to secrete inflammatory cytokines and inhibit apoptosis. Int J Clin Exp Med 8:17530–17535

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Marchesini MI, Herrmann CK, Salcedo SP, Gorvel JP, Comerci DJ (2011) In search of Brucella abortus type IV secretion substrates: Screening and identification of four proteins translocated into host cells through virb system. Cell Microbiol 13:1261–1274

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. McQuiston JR, Schurig GG, Sriranganathan N, Boyle SM (1995) Transformation of Brucella species with suicide and broad host-range plasmids. Methods Mol Biol 47:143–148

    CAS  PubMed  Google Scholar 

  20. Nambiar PR, Kirchain SM, Courmier K, Xu S, Taylor NS, Theve EJ, Patterson MM, Fox JG (2006) Progressive proliferative and dysplastic typhlocolitis in aging syrian hamsters naturally infected with helicobacter spp.: A spontaneous model of inflammatory bowel disease. Vet Pathol 43:2–14

    Article  CAS  PubMed  Google Scholar 

  21. Wang Y, Bai Y, Qu Q, Xu J, Chen Y, Zhong Z, Qiu Y, Wang T, Du X, Wang Z, Yu S, Fu S, Yuan J, Zhen Q, Yu Y, Chen Z, Huang L (2011) The 16mδvjbr as an ideal live attenuated vaccine candidate for differentiation between Brucella vaccination and infection. Vet Microbiol 151:354–362

    Article  CAS  PubMed  Google Scholar 

  22. Wang Y, Chen Z, Qiao F, Ying T, Yuan J, Zhong Z, Zhou L, Du X, Wang Z, Zhao J, Dong S, Jia L, Yuan X, Yang R, Sun Y, Huang L (2009) Comparative proteomics analyses reveal the virb of b. Melitensis affects expression of intracellular survival related proteins. PLoS One 4:e5368

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Wang Y, Chen Z, Qiao F, Zhong Z, Xu J, Wang Z, Du X, Qu Q, Yuan J, Jia L, Song H, Sun Y, Huang L (2010) The type IV secretion system affects the expression of omp25/omp31 and the outer membrane properties of Brucella melitensis. FEMS Microbiol Lett 303:92–100

    Article  CAS  PubMed  Google Scholar 

  24. Yuan M, Li Y, Zhong C, Li Y, Niu J, Gong J (2015) Overexpression of neuritin in gastric cancer. Oncol Lett 10:3832–3836

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Zhang H, Dou X, Li Z, Zhang Y, Zhang J, Guo F, Wang Y, Wang Z, Li T, Gu X, Chen C (2016) Expression and regulation of the ery operon of Brucella melitensis in human trophoblast cells. Exp Ther Med 12:2723–2728

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Zhang Y, Li T, Zhang J, Li Z, Zhang Y, Wang Z, Feng H, Wang Y, Chen C, Zhang H (2016) The Brucella melitensis m5-90 phosphoglucomutase (pgm) mutant is attenuated and confers protection against wild-type challenge in balb/c mice. World J Microbiol Biotechnol 32:58

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was supported by grants from the National key Research and Development Program of China (2017YFD0500304), the Training Program for Excellent Young Teachers Colleges and Universities of Corps (CZ027202), the Youth Science and Technology Innovation Leading Talent Program of Corps (2017CB002), and the National Natural Science Foundation of China (31460650, 31602080).

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Correspondence to Yayin Qi or Hui Zhang.

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Zhang, J., Li, M., Li, Z. et al. Deletion of the Type IV Secretion System Effector VceA Promotes Autophagy and Inhibits Apoptosis in Brucella-Infected Human Trophoblast Cells. Curr Microbiol 76, 510–519 (2019). https://doi.org/10.1007/s00284-019-01651-6

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