Neutrophil elastase targets virulence factors of enterobacteria


Shigellae cause bacillary dysentery, a bloody form of diarrhoea that affects almost 200 million people and causes nearly 2 million deaths per year1. Shigella invades the colonic mucosa, where it initiates an acute inflammation, rich in neutrophils, that initially contributes to tissue damage and eventually resolves the infection2. Neutrophils are phagocytic cells that kill microorganisms3,4 but it is unclear how neutrophils control pathogenic bacteria expressing virulence factors that manipulate host cells. In contrast to other cells, neutrophils prevent the escape of Shigella from phagocytic vacuoles in which the bacteria are killed5. Here we identify human neutrophil elastase (NE) as a key host defence protein: NE degrades Shigella virulence factors at a 1,000-fold lower concentration than that needed to degrade other bacterial proteins. In neutrophils in which NE is inactivated pharmacologically or genetically, Shigella escapes from phagosomes, increasing bacterial survival. NE also preferentially cleaves virulence factors of Salmonella and Yersinia. These findings establish NE as the first neutrophil factor that targets bacterial virulence proteins.

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Figure 1: Human neutrophil extract enriched in granule proteins (hNEGP) targets virulence proteins of Shigella.
Figure 2: NE degrades Shigella virulence proteins.
Figure 3: NE degrades secreted virulence proteins of Gram-negative bacterial pathogens.
Figure 4: Abrogation of NE permits Shigella to escape the phagosome of neutrophils.


  1. 1

    Kotloff, K. L. et al. Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. Bull. World Health Org. 77, 651–666 (1999).

    CAS  PubMed  Google Scholar 

  2. 2

    Perdomo, O. J. et al. Acute inflammation causes epithelial invasion and mucosal destruction in experimental shigellosis. J. Exp. Med. 180, 1307–1319 (1994).

    CAS  Article  Google Scholar 

  3. 3

    Klebanoff, S. J. in Inflammation: Basic Principles and Clinical Correlates (eds Gallin, J. I. & Snyderman, R.) 721–768 (Lippincott Williams & Wilkins, Philadelphia, 1999).

    Google Scholar 

  4. 4

    Elsbach, P., Weiss, J. & Levy, O. in Inflammation: Basic Principles and Clinical Correlates (eds Gallin, J. J. & Snyderman, R.) 801–817 (Lippincott Williams & Wilkins, Philadelphia, 1999).

    Google Scholar 

  5. 5

    Mandic-Mulec, I., Weiss, J. & Zychlinsky, A. Shigella flexneri is trapped in polymorphnuclear leukocyte vacuoles and efficiently killed. Infect. Immun. 65, 110–115 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  6. 6

    Hale, T. L. Genetic basis of virulence in Shigella species. Microbiol. Rev. 55, 206–224 (1991).

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7

    Cornelis, G. R. & Van Gijsegem, F. Assembly and function of type III secretory systems. Annu. Rev. Microbiol. 54, 735–774 (2000).

    CAS  Article  Google Scholar 

  8. 8

    Suzuki, T., Lett, M. C. & Sasakawa, C. Extracellular transport of VirG protein in Shigella. J. Biol. Chem. 270, 30874–30880 (1995).

    CAS  Article  Google Scholar 

  9. 9

    Bahrani, F. K., Sansonetti, P. J. & Parsot, C. Secretion of Ipa proteins by Shigella flexneri: inducer molecules and kinetics of activation. Infect. Immun. 65, 4005–4010 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. 10

    Owen, C. A. & Campbell, E. J. The cell biology of leukocyte-mediated proteolysis. J. Leukoc. Biol. 65, 137–150 (1999).

    CAS  Article  Google Scholar 

  11. 11

    Campbell, E. J., Silverman, E. K. & Campbell, M. A. Elastase and cathepsin G of human monocytes. Quantification of cellular content, release in response to stimuli, and heterogeneity in elastase-mediated proteolytic activity. J. Immunol. 143, 2961–2968 (1989).

    CAS  PubMed  Google Scholar 

  12. 12

    Veale, C. A. et al. Orally active trifluoromethyl ketone inhibitors of human leukocyte elastase. J. Med. Chem. 40, 3173–3181 (1997).

    CAS  Article  Google Scholar 

  13. 13

    Wright, C. D., Kennedy, J. A., Zitnik, R. J. & Kashem, M. A. Inhibition of murine neutrophil serine proteinases by human and murine secretory leukocyte protease inhibitor. Biochem. Biophys. Res. Commun. 254, 614–617 (1999).

    CAS  Article  Google Scholar 

  14. 14

    Belaaouaj, A., Kim, K. S. & Shapiro, S. D. Degradation of outer membrane protein A in Escherichia coli killing by neutrophil elastase. Science 289, 1185–1188 (2000).

    ADS  CAS  Article  Google Scholar 

  15. 15

    Panyutich, A., Shi, J., Boutz, P. L., Zhao, C. & Ganz, T. Porcine polymorphonuclear leukocytes generate extracellular microbicidal activity by elastase-mediated activation of secreted proprotegrins. Infect. Immun. 65, 978–985 (1997).

    CAS  PubMed  PubMed Central  Google Scholar 

  16. 16

    Tamano, K. et al. Supramolecular structure of the Shigella type III secretion machinery: the needle part is changeable in length and essential for delivery of effectors. EMBO J. 19, 3876–3887 (2000).

    CAS  Article  Google Scholar 

  17. 17

    Blocker, A. et al. Structure and composition of the Shigella flexneri ‘needle complex’, a part of its type III secreton. Mol. Microbiol. 39, 652–663 (2001).

    CAS  Article  Google Scholar 

  18. 18

    Hayashi, F. et al. The innate immune response to bacterial flagellin is mediated by Toll-like receptor 5. Nature 410, 1099–1103 (2001).

    ADS  CAS  Article  Google Scholar 

  19. 19

    Eisenhauer, P. B., Harwig, S. S. & Lehrer, R. I. Cryptdins: Antimicrobial defensins of the murine small intestine. Infect. Immun. 60, 3556–3565 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  20. 20

    Francois, M., Le Cabec, V., Dupont, M. A., Sansonetti, P. J. & Maridonneau-Parini, I. Induction of necrosis in human neutrophils by Shigella flexneri requires type III secretion, IpaB and IpaC invasins, and actin polymerization. Infect. Immun. 68, 1289–1296 (2000).

    CAS  Article  Google Scholar 

  21. 21

    Pallen, M. J., Dougan, G. & Frankel, G. Coiled-coil domains in proteins secreted by type III secretion systems. Mol. Microbiol. 25, 423–425 (1997).

    CAS  Article  Google Scholar 

  22. 22

    Miao, E. A. et al. leucine-rich repeat proteins are targeted to the SPI1 and SPI2 type III secretion systems. Mol. Microbiol. 34, 850–864 (1999).

    CAS  Article  Google Scholar 

  23. 23

    Menard, R., Sansonetti, P. J. & Parsot, C. Nonpolar mutagenesis of the ipa genes defines IpaB, IpaC, and IpaD as effectors of Shigella flexneri entry into epithelial cells. J. Bacteriol. 175, 5899–5906 (1993).

    CAS  Article  Google Scholar 

  24. 24

    Weiss, J., Elsbach, P., Olsson, I. & Odeberg, H. Purification and characterization of a potent bactericidal and membrane active protein from the granules of human polymorphonucelar leukocytes. J. Biol. Chem. 253, 2664–2672 (1978).

    CAS  PubMed  Google Scholar 

  25. 25

    Lee, V. T. & Schneewind, O. Type III machines of pathogenic Yersiniae secrete virulence factors into the extracellular milieu. Mol. Microbiol. 31, 1619–1629 (1999).

    CAS  Article  Google Scholar 

  26. 26

    Ginocchio, C. C. & Galan, J. E. Functional conservation among members of the Salmonella typhimurium InvA family of proteins. Infect. Immun. 63, 729–732 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. 27

    Huang, Y. I. et al. Effect of trifluoromethyl ketone-based elastase inhibitors on neutrophil function in vitro. J. Leukoc. Biol. 64, 322–330 (1998).

    CAS  Article  Google Scholar 

  28. 28

    Weiss, J., Kao, L., Victor, M. & Elsbach, P. Oxygen-independent intracellular and oxygen-dependent extracellular killing of Escherichia coli S15 by human polymorphonuclear leukocytes. J. Clin. Invest. 76, 206–212 (1985).

    CAS  Article  Google Scholar 

  29. 29

    Sansonetti, P. J., Arondel, J., Huerre, M., Harada, A. & Matsushima, K. Interleukin-8 controls bacterial transepithelial translocation at the cost of epithelial destruction in experimental shigellosis. Infect. Immun. 67, 1471–1480 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  30. 30

    Gbarah, A. et al. Shigella flexneri transformants expressing type 1 (mannose-specific) fimbriae bind to, activate, and are killed by phagocytic cells. Infect. Immun. 61, 1687–1693 (1993).

    CAS  PubMed  PubMed Central  Google Scholar 

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We thank D. Weiss and C. Scharff for critical reading of the manuscript, T. Nubert and D. Wu for performing MS analysis, and staff of the Rockefeller University EM facility. A.Z. is a recipient of the Irma T. Hirschl Trust Career Scientist Award. Financial support from the National Institutes of Health is acknowledged.

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Correspondence to Arturo Zychlinsky.

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Weinrauch, Y., Drujan, D., Shapiro, S. et al. Neutrophil elastase targets virulence factors of enterobacteria. Nature 417, 91–94 (2002).

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