Skip to main content
Springer Nature Link
Log in

Expression of PE_PGRS 62 protein in Mycobacterium smegmatis decrease mRNA expression of proinflammatory cytokines IL-1β, IL-6 in macrophages

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The pathogenesis of tuberculosis causing Mycobacterium bovis is largely due to its successful entry and survival in macrophages. Previous research indicated that mycobacteria-specific PE_PGRS genes code for cell surface proteins which may have role in mediating interactions with macrophages. In this study, we expressed PE_PGRS 62 gene in a non-pathogenic fast growing Mycobacterium smegmatis strain and found that the recombinant Mycobacterium smegmatis decreased macrophages livability in a dosage-dependent manner and time-dependent manner, compared with parental strain containing the vector only. To explore whether PE_PGRS 62 modulates the gene expression profile of macrophages, we stimulated macrophages by the M. smegmatis strain expressing PE_PGRS 62 as well as the control strains, followed by real-time RT–PCR assay for the mRNA expression level of IL-1β, IL-6, and iNOS. The results showed that the expression of IL-1β, IL-6 in macrophages were down-regulated by stimulation with the M. smegmatis strain expressing PE_PGRS 62 compared to the control strains (P < 0.05). In contrast, there were no measurable differences in the expression of iNOS. Overall, we demonstrated that PE_PGRS 62 protein altered the immune environment of the host cells, which suggest that the pathogenic PE_PGRS 62 protein altering the immune mechanism maybe involved in the pathogenesis of mycobacterial disease.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Canada)

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. de la Rua-Domenech R (2006) Human Mycobacterium bovis infection in the United Kingdom: incidence, risks, control measures and review of the zoonotic aspects of bovine tuberculosis. Tuberculosis (Edinb) 86:77–109

    Article  Google Scholar 

  2. O’Reilly LM, Daborn CJ (1995) The epidemiology of Mycobacterium bovis infections in animals and man: a review. Tuber Lung Dis 76(Suppl 1):1–46

    Article  PubMed  Google Scholar 

  3. Monack DM, Mueller A, Falkow S (2004) Persistent bacterial infections: the interface of the pathogen and the host immune system. Nat Rev Microbiol 2:747–765

    Article  CAS  PubMed  Google Scholar 

  4. Stewart GR, Robertson BD, Young DB (2003) Tuberculosis: a problem with persistence. Nat Rev Microbiol 1:97–105

    Article  CAS  PubMed  Google Scholar 

  5. Glickman MS, Jacobs WR Jr (2001) Microbial pathogenesis of Mycobacterium tuberculosis: dawn of a discipline. Cell 104:477–485

    Article  CAS  PubMed  Google Scholar 

  6. Cole ST, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D, Gordon SV, Eiglmeier K, Gas S, Barry CE 3rd, Tekaia F, Badcock K, Basham D, Brown D, Chillingworth T, Connor R, Davies R, Devlin K, Feltwell T, Gentles S, Hamlin N, Holroyd S, Hornsby T, Jagels K, Krogh A, McLean J, Moule S, Murphy L, Oliver K, Osborne J, Quail MA, Rajandream MA, Rogers J, Rutter S, Seeger K, Skelton J, Squares R, Squares S, Sulston JE, Taylor K, Whitehead S, Barrell BG (1998) Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature 393:537–544

    Article  CAS  PubMed  Google Scholar 

  7. Fleischmann RD, Alland D, Eisen JA, Carpenter L, White O, Peterson J, DeBoy R, Dodson R, Gwinn M, Haft D, Hickey E, Kolonay JF, Nelson WC, Umayam LA, Ermolaeva M, Salzberg SL, Delcher A, Utterback T, Weidman J, Khouri H, Gill J, Mikula A, Bishai W, Jacobs WR Jr, Venter JC Jr, Fraser CM (2002) Whole-genome comparison of Mycobacterium tuberculosis clinical and laboratory strains. J Bacteriol 184:5479–5490

    Article  CAS  PubMed  Google Scholar 

  8. Garnier T, Eiglmeier K, Camus JC, Medina N, Mansoor H, Pryor M, Duthoy S, Grondin S, Lacroix C, Monsempe C, Simon S, Harris B, Atkin R, Doggett J, Mayes R, Keating L, Wheeler PR, Parkhill J, Barrell BG, Cole ST, Gordon SV, Hewinson RG (2003) The complete genome sequence of Mycobacterium bovis. Proc Natl Acad Sci USA 100:7877–7882

    Article  CAS  PubMed  Google Scholar 

  9. Voskuil MI, Schnappinger D, Rutherford R, Liu Y, Schoolnik GK (2004) Regulation of the Mycobacterium tuberculosis PE/PPE genes. Tuberculosis (Edinb) 84:256–262

    Article  CAS  Google Scholar 

  10. Brennan MJ, Delogu G (2002) The PE multigene family: a ‘molecular mantra’ for mycobacteria. Trends Microbiol 10:246–249

    Article  CAS  PubMed  Google Scholar 

  11. Dheenadhayalan V, Delogu G, Brennan MJ (2006) Expression of the PE_PGRS 33 protein in Mycobacterium smegmatis triggers necrosis in macrophages and enhanced mycobacterial survival. Microbes Infect 8:262–272

    Article  CAS  PubMed  Google Scholar 

  12. Delogu G, Pusceddu C, Bua A, Fadda G, Brennan MJ, Zanetti S (2004) Rv1818c-encoded PE_PGRS protein of Mycobacterium tuberculosis is surface exposed and influences bacterial cell structure. Mol Microbiol 52:725–733

    Article  CAS  PubMed  Google Scholar 

  13. Banu S, Honore N, Saint-Joanis B, Philpott D, Prevost MC, Cole ST (2002) Are the PE-PGRS proteins of Mycobacterium tuberculosis variable surface antigens? Mol Microbiol 44:9–19

    Article  CAS  PubMed  Google Scholar 

  14. Brennan MJ, Delogu G, Chen Y, Bardarov S, Kriakov J, Alavi M, Jacobs WR Jr (2001) Evidence that mycobacterial PE_PGRS proteins are cell surface constituents that influence interactions with other cells. Infect Immun 69:7326–7333

    Article  CAS  PubMed  Google Scholar 

  15. Chaitra MG, Hariharaputran S, Chandra NR, Shaila MS, Nayak R (2005) Defining putative T cell epitopes from PE and PPE families of proteins of Mycobacterium tuberculosis with vaccine potential. Vaccine 23:1265–1272

    Article  CAS  PubMed  Google Scholar 

  16. Basu S, Pathak SK, Banerjee A, Pathak S, Bhattacharyya A, Yang Z, Talarico S, Kundu M, Basu J (2007) Execution of macrophage apoptosis by PE_PGRS33 of Mycobacterium tuberculosis is mediated by Toll-like receptor 2-dependent release of tumor necrosis factor-alpha. J Biol Chem 282:1039–1050

    Article  CAS  PubMed  Google Scholar 

  17. Delogu G, Sanguinetti M, Pusceddu C, Bua A, Brennan MJ, Zanetti S, Fadda G (2006) PE_PGRS proteins are differentially expressed by Mycobacterium tuberculosis in host tissues. Microbes Infect 8:2061–2067

    Article  CAS  PubMed  Google Scholar 

  18. Dheenadhayalan V, Delogu G, Sanguinetti M, Fadda G, Brennan MJ (2006) Variable expression patterns of Mycobacterium tuberculosis PE_PGRS genes: evidence that PE_PGRS16 and PE_PGRS26 are inversely regulated in vivo. J Bacteriol 188:3721–3725

    Article  CAS  PubMed  Google Scholar 

  19. Betts JC, Lukey PT, Robb LC, McAdam RA, Duncan K (2002) Evaluation of a nutrient starvation model of Mycobacterium tuberculosis persistence by gene and protein expression profiling. Mol Microbiol 43:717–731

    Article  CAS  PubMed  Google Scholar 

  20. Ramakrishnan L, Federspiel NA, Falkow S (2000) Granuloma-specific expression of Mycobacterium virulence proteins from the glycine-rich PE-PGRS family. Science 288:1436–1439

    Article  CAS  PubMed  Google Scholar 

  21. Xu G, Li Y, Yang J, Zhou X, Yin X, Liu M, Zhao D (2007) Effect of recombinant Mce4A protein of Mycobacterium bovis on expression of TNF-alpha, iNOS, IL-6, and IL-12 in bovine alveolar macrophages. Mol Cell Biochem 302:1–7

    Article  CAS  PubMed  Google Scholar 

  22. Snapper SB, Melton RE, Mustafa S, Kieser T, Jacobs WR Jr (1990) Isolation and characterization of efficient plasmid transformation mutants of Mycobacterium smegmatis. Mol Microbiol 4:1911–1919

    Article  CAS  PubMed  Google Scholar 

  23. Stover C-K, Cruz V-F, Fuerst T-R (1991) New use of BCG for recombinant vaccines. Nature 351:456–460

    Article  CAS  PubMed  Google Scholar 

  24. Master SS, Rampini SK, Davis AS, Keller C, Ehlers S, Springer B, Timmins GS, Sander P, Deretic V (2008) Mycobacterium tuberculosis prevents inflammasome activation. Cell Host Microbe 3:224–232

    Article  CAS  PubMed  Google Scholar 

  25. Saunders BM, Cooper AM (2000) Restraining mycobacteria: role of granulomas in mycobacterial infections. Immunol Cell Biol 78:334–341

    Article  CAS  PubMed  Google Scholar 

  26. Hirano T, Taga T, Nakano N, Yasukawa K, Kashiwamura S, Shimizu K, Nakajima K, Pyun KH, Kishimoto T (1985) Purification to homogeneity and characterization of human B-cell differentiation factor (BCDF or BSFp-2). Proc Natl Acad Sci USA 82:5490–5494

    Article  CAS  PubMed  Google Scholar 

  27. Corbel C, Melchers F (1984) The synergism of accessory cells and of soluble alpha-factors derived from them in the activation of B cells to proliferation. Immunol Rev 78:51–74

    Article  CAS  PubMed  Google Scholar 

  28. Weissenbach J, Chernajovsky Y, Zeevi M, Shulman L, Soreq H, Nir U, Wallach D, Perricaudet M, Tiollais P, Revel M (1980) Two interferon mRNAs in human fibroblasts: in vitro translation and Escherichia coli cloning studies. Proc Natl Acad Sci USA 77:7152–7156

    Article  CAS  PubMed  Google Scholar 

  29. Pasare C, Medzhitov R (2003) Toll pathway-dependent blockade of CD4+CD25+ T cell-mediated suppression by dendritic cells. Science 299:1033–1036

    Article  CAS  PubMed  Google Scholar 

  30. Fang FC (1997) Perspectives series: host/pathogen interactions. Mechanisms of nitric oxide-related antimicrobial activity. J Clin Invest 99:2818–2825

    Article  CAS  PubMed  Google Scholar 

  31. John D. MacMicking RJN, LaCourse R, Mudgett JS, Shah SK, Nathan CF (1997) Identification of nitric oxide synthase as a protective locus against tuberculosis. Proc Natl Acad Sci USA 94: 5243-5248

    Google Scholar 

  32. Chan J, Xing Y, Magliozzo RS, Bloom BR (1992) Killing of virulent Mycobacterium tuberculosis by reactive nitrogen intermediates produced by activated murine macrophages. J Exp Med 175:1111–1122

    Article  CAS  PubMed  Google Scholar 

  33. Campuzano J, Aguilar D, Arriaga K, Leon JC, Salas-Rangel LP, Gonzalez-y-Merchand J, Hernandez-Pando R, Espitia C (2007) The PGRS domain of Mycobacterium tuberculosis PE_PGRS Rv1759c antigen is an efficient subunit vaccine to prevent reactivation in a murine model of chronic tuberculosis. Vaccine 25:3722–3729

    Article  CAS  PubMed  Google Scholar 

  34. Cockle PJ, Gordon SV, Lalvani A, Buddle BM, Hewinson RG, Vordermeier HM (2002) Identification of novel Mycobacterium tuberculosis antigens with potential as diagnostic reagents or subunit vaccine candidates by comparative genomics. Infect Immun 70:6996–7003

    Article  CAS  PubMed  Google Scholar 

  35. Chaitra MG, Nayak R, Shaila MS (2007) Modulation of immune responses in mice to recombinant antigens from PE and PPE families of proteins of Mycobacterium tuberculosis by the Ribi adjuvant. Vaccine 25:7168–7176

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by Ministry of Agriculture Key Program, China (Project No. 2009ZX08008-010B), Ministry of Agriculture Key Program, China (Project No. 2009ZX08007-008B), P Ministry of Agriculture Key Program, China (Project No. 2009ZX08009-183B), Natural Science Foundation of China (Project No. 30871854) and National Science and Technology Supporting Program of China (Project No. 2006BAD06A13).

Author information

Authors and Affiliations

  1. National Animal Transmissible Spongiform Encephalopathy Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, People’s Republic of China

    Ying Huang, Yang Wang, Yu Bai, Lifeng Yang & Deming Zhao

  2. China Animal Disease Control Center, Beijing, 100125, People’s Republic of China

    Zhi Gang Wang

Authors
  1. Ying Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yu Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhi Gang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Lifeng Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Deming Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Deming Zhao.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Y., Wang, Y., Bai, Y. et al. Expression of PE_PGRS 62 protein in Mycobacterium smegmatis decrease mRNA expression of proinflammatory cytokines IL-1β, IL-6 in macrophages. Mol Cell Biochem 340, 223–229 (2010). https://doi.org/10.1007/s11010-010-0421-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-010-0421-x

Keywords