Quorum Sensing pp 133-145 | Cite as

Modulation of Mammalian Cell Processes by Bacterial Quorum Sensing Molecules

  • Vladimir V. Kravchenko
  • Richard J. Ulevitch
  • Gunnar F. Kaufmann
Part of the Methods in Molecular Biology book series (MIMB, volume 692)


Microbial pathogens use a wide repertoire of pathogen-associated molecular patterns (PAMPs) that affect host cell responses through activation of intracellular signaling events in a PAMP-specific manner. Here we describe a set of western blot-based methodologies for the evaluation of biochemical effects specifically induced by N-(3-oxo-acyl) homoserine lactones (3-oxo-AHLs) small molecules secreted by a number of Gram-negative bacteria, including the opportunistic human pathogen Pseudomonas aeruginosa. First, we will highlight the AHL-mediated effects on proapoptotic and stress pathways. Secondly, we will demonstrate that AHLs possess the ability to alter stimulus-induced NF-κB signaling, a key biochemical marker of inflammation and innate immune responses.

Key words

Acyl homoserine lactones Macrophages NF-kappa B Pseudomonas aeruginosa MAP kinase Apoptosis 



The authors would like to thank Colin A. Lowery for critical reading the manuscript. This work was supported by the National Institutes of Health (AI079436 to VVK and AI080715 to GFK). This is manuscript number 20550 from The Scripps Research Institute.


  1. 1.
    Janeway, C. A., Jr., and Medzhitov, R. (2002) Innate immune recognition, Annu Rev Immunol 20, 197–216.PubMedCrossRefGoogle Scholar
  2. 2.
    Hoffmann, A., and Baltimore, D. (2006) Circuitry of nuclear factor kappaB signaling, Immunol Rev 210, 171–186.PubMedCrossRefGoogle Scholar
  3. 3.
    Schmitz, M. L., and de la Vega, L. (2008) A bacterial small molecule undermining immune response signaling, Chembiochem 9, 2575–2577.PubMedCrossRefGoogle Scholar
  4. 4.
    DiMango, E., Ratner, A. J., Bryan, R., Tabibi, S., and Prince, A. (1998) Activation of NF-kappaB by adherent Pseudomonas aeruginosa in normal and cystic fibrosis respiratory epithelial cells, J Clin Invest 101, 2598–2605.PubMedCrossRefGoogle Scholar
  5. 5.
    Pearson, J. P., Gray, K. M., Passador, L., Tucker, K. D., Eberhard, A., Iglewski, B. H., and Greenberg, E. P. (1994) Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes, Proc Natl Acad Sci U S A 91, 197–201.PubMedCrossRefGoogle Scholar
  6. 6.
    Smith, R. S., and Iglewski, B. H. (2003) Pseudomonas aeruginosa quorum sensing as a potential antimicrobial target, J Clin Invest 112, 1460–1465.PubMedGoogle Scholar
  7. 7.
    Shiner, E. K., Rumbaugh, K. P., and Williams, S. C. (2005) Inter-kingdom signaling: deciphering the language of acyl homoserine lactones, FEMS Microbiol Rev 29, 935–947.PubMedCrossRefGoogle Scholar
  8. 8.
    DiMango, E., Zar, H. J., Bryan, R., and Prince, A. (1995) Diverse Pseudomonas aeruginosa gene products stimulate respiratory epithelial cells to produce interleukin-8, J Clin Invest 96, 2204–2210.PubMedCrossRefGoogle Scholar
  9. 9.
    Palfreyman, R. W., Watson, M. L., Eden, C., and Smith, A. W. (1997) Induction of biologically active interleukin-8 from lung epithelial cells by Burkholderia (Pseudomonas) cepacia products, Infect Immun 65, 617–622.PubMedGoogle Scholar
  10. 10.
    Telford, G., Wheeler, D., Williams, P., Tomkins, P. T., Appleby, P., Sewell, H., Stewart, G. S., Bycroft, B. W., and Pritchard, D. I. (1998) The Pseudomonas aeruginosa quorum-sensing signal molecule N-(3-oxododecanoyl)-l-homoserine lactone has immunomodulatory activity, Infect Immun 66, 36–42.PubMedGoogle Scholar
  11. 11.
    Smith, R. S., Fedyk, E. R., Springer, T. A., Mukaida, N., Iglewski, B. H., and Phipps, R. P. (2001) IL-8 production in human lung fibroblasts and epithelial cells activated by the Pseudomonas autoinducer N-3-oxododecanoyl homoserine lactone is transcriptionally regulated by NF-kappa B and activator protein-2, J Immunol 167, 366–374.PubMedGoogle Scholar
  12. 12.
    Tateda, K., Ishii, Y., Horikawa, M., Matsumoto, T., Miyairi, S., Pechere, J. C., Standiford, T. J., Ishiguro, M., and Yamaguchi, K. (2003) The Pseudomonas aeruginosa autoinducer N-3-oxododecanoyl homoserine lactone accelerates apoptosis in macrophages and neutrophils, Infect Immun 71, 5785–5793.PubMedCrossRefGoogle Scholar
  13. 13.
    Chhabra, S. R., Harty, C., Hooi, D. S., Daykin, M., Williams, P., Telford, G., Pritchard, D. I., and Bycroft, B. W. (2003) Synthetic analogues of the bacterial signal (quorum sensing) molecule N-(3-oxododecanoyl)-l-homoserine lactone as immune modulators, J Med Chem 46, 97–104.PubMedCrossRefGoogle Scholar
  14. 14.
    Ritchie, A. J., Yam, A. O., Tanabe, K. M., Rice, S. A., and Cooley, M. A. (2003) Modification of in vivo and in vitro T- and B-cell-mediated immune responses by the Pseudomonas aeruginosa quorum-sensing molecule N-(3-oxododecanoyl)-l-homoserine lactone, Infect Immun 71, 4421–4431.PubMedCrossRefGoogle Scholar
  15. 15.
    Li, L., Hooi, D., Chhabra, S. R., Pritchard, D., and Shaw, P. E. (2004) Bacterial N-acylhomoserine lactone-induced apoptosis in breast carcinoma cells correlated with down-modulation of STAT3, Oncogene 23, 4894–4902.PubMedCrossRefGoogle Scholar
  16. 16.
    Shiner, E. K., Terentyev, D., Bryan, A., Sennoune, S., Martinez-Zaguilan, R., Li, G., Gyorke, S., Williams, S. C., and Rumbaugh, K. P. (2006) Pseudomonas aeruginosa autoinducer modulates host cell responses through calcium signalling, Cell Microbiol 8, 1601–1610.PubMedCrossRefGoogle Scholar
  17. 17.
    Kravchenko, V. V., Kaufmann, G. F., Mathison, J. C., Scott, D. A., Katz, A. Z., Wood, M. R., Brogan, A. P., Lehmann, M., Mee, J. M., Iwata, K., Pan, Q., Fearns, C., Knaus, U. G., Meijler, M. M., Janda, K. D., and Ulevitch, R. J. (2006) N-(3-oxo-acyl)homoserine lactones signal cell activation through a mechanism distinct from the canonical pathogen-associated molecular pattern recognition receptor pathways, J Biol Chem 281, 28822–28830.PubMedCrossRefGoogle Scholar
  18. 18.
    Jahoor, A., Patel, R., Bryan, A., Do, C., Krier, J., Watters, C., Wahli, W., Li, G., Williams, S. C., and Rumbaugh, K. P. (2008) Peroxisome proliferator-activated receptors mediate host cell proinflammatory responses to Pseudomonas aeruginosa autoinducer, J Bacteriol 190, 4408–4415.PubMedCrossRefGoogle Scholar
  19. 19.
    Cooley, M., Chhabra, S. R., and Williams, P. (2008) N-Acylhomoserine lactone-mediated quorum sensing: a twist in the tail and a blow for host immunity, Chem Biol 15, 1141–1147.PubMedCrossRefGoogle Scholar
  20. 20.
    Williams, S. C., Patterson, E. K., Carty, N. L., Griswold, J. A., Hamood, A. N., and Rumbaugh, K. P. (2004) Pseudomonas aeruginosa autoinducer enters and functions in mammalian cells, J Bacteriol 186, 2281–2287.PubMedCrossRefGoogle Scholar
  21. 21.
    Ritchie, A. J., Whittall, C., Lazenby, J. J., Chhabra, S. R., Pritchard, D. I., and Cooley, M. A. (2007) The immunomodulatory Pseudomonas aeruginosa signalling molecule N-(3-oxododecanoyl)-l-homoserine lactone enters mammalian cells in an unregulated fashion, Immunol Cell Biol 85, 596–602.PubMedCrossRefGoogle Scholar
  22. 22.
    Yates, E. A., Philipp, B., Buckley, C., Atkinson, S., Chhabra, S. R., Sockett, R. E., Goldner, M., Dessaux, Y., Camara, M., Smith, H., and Williams, P. (2002) N-acylhomoserine lactones undergo lactonolysis in a pH-, temperature-, and acyl chain length-dependent manner during growth of Yersinia pseudotuberculosis and Pseudomonas aeruginosa, Infect Immun 70, 5635–5646.PubMedCrossRefGoogle Scholar
  23. 23.
    Kaufmann, G. F., Sartorio, R., Lee, S. H., Rogers, C. J., Meijler, M. M., Moss, J. A., Clapham, B., Brogan, A. P., Dickerson, T. J., and Janda, K. D. (2005) Revisiting quorum sensing: discovery of additional chemical and biological functions for 3-oxo-N-acylhomoserine lactones, Proc Natl Acad Sci U S A 102, 309–314.PubMedCrossRefGoogle Scholar
  24. 24.
    Ozer, E. A., Pezzulo, A., Shih, D. M., Chun, C., Furlong, C., Lusis, A. J., Greenberg, E. P., and Zabner, J. (2005) Human and murine paraoxonase 1 are host modulators of Pseudomonas aeruginosa quorum-sensing, FEMS Microbiol Lett 253, 29–37.PubMedCrossRefGoogle Scholar
  25. 25.
    Kravchenko, V. V., Kaufmann, G. F., Mathison, J. C., Scott, D. A., Katz, A. Z., Grauer, D. C., Lehmann, M., Meijler, M. M., Janda, K. D., and Ulevitch, R. J. (2008) Modulation of gene expression via disruption of NF-kappaB signaling by a bacterial small molecule, Science 321, 259–263.PubMedCrossRefGoogle Scholar
  26. 26.
    Charlton, T. S., de Nys, R., Netting, A., Kumar, N., Hentzer, M., Givskov, M., and Kjelleberg, S. (2000) A novel and sensitive method for the quantification of N-3-oxoacyl homoserine lactones using gas chromatography–mass spectrometry: application to a model bacterial biofilm, Environ Microbiol 2, 530–541.PubMedCrossRefGoogle Scholar
  27. 27.
    Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227, 680–685.PubMedCrossRefGoogle Scholar
  28. 28.
    Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, Vol. 3, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Vladimir V. Kravchenko
    • 1
  • Richard J. Ulevitch
    • 1
  • Gunnar F. Kaufmann
    • 2
  1. 1.Department of Immunology & Microbial ScienceThe Scripps Research InstituteLa JollaUSA
  2. 2.Departments of Chemistry and Immunology & Microbial ScienceThe Scripps Research InstituteLa JollaUSA

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