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Macrophage elastase kills bacteria within murine macrophages

Abstract

Macrophages are aptly positioned to function as the primary line of defence against invading pathogens in many organs, including the lung and peritoneum. Their ability to phagocytose and clear microorganisms has been well documented1,2. Macrophages possess several substances with which they can kill bacteria, including reactive oxygen species, nitric oxide, and antimicrobial proteins3,4,5,6,7,8,9. We proposed that macrophage-derived proteinases may contribute to the antimicrobial properties of macrophages. Macrophage elastase (also known as matrix metalloproteinase 12 or MMP12) is an enzyme predominantly expressed in mature tissue macrophages10 and is implicated in several disease processes, including emphysema11. Physiological functions for MMP12 have not been described. Here we show that Mmp12-/- mice exhibit impaired bacterial clearance and increased mortality when challenged with both Gram-negative and Gram-positive bacteria at macrophage-rich portals of entry, such as the peritoneum and lung. Intracellular stores of MMP12 are mobilized to macrophage phagolysosomes after the ingestion of bacterial pathogens. Once inside phagolysosomes, MMP12 adheres to bacterial cell walls where it disrupts cellular membranes resulting in bacterial death. The antimicrobial properties of MMP12 do not reside within its catalytic domain, but rather within the carboxy-terminal domain. This domain contains a unique four amino acid sequence on an exposed β loop of the protein that is required for the observed antimicrobial activity. The present study represents, to our knowledge, the first report of direct antimicrobial activity by a matrix metallopeptidase, and describes a new antimicrobial peptide that is sequentially and structurally unique in nature.

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Figure 1: MMP12 provides survival advantage in Gram-positive and Gram-negative infection.
Figure 2: MMP12 improves bacterial clearance and survival in S. aureus pneumonia.
Figure 3: MMP12 is required for early intracellular bacterial killing by macrophages.
Figure 4: MMP12 CTD possesses bactericidal activity.

References

  1. 1

    Green, G. M. & Kass, E. H. The role of the alveolar macrophage in the clearance of bacteria from the lung. J. Exp. Med. 119, 167–176 (1964)

    CAS  Article  Google Scholar 

  2. 2

    Gordon, S. The macrophage: Past, present and future. Eur. J. Immunol. 37, S9–S17 (2007)

    CAS  Article  Google Scholar 

  3. 3

    Shiloh, M. U. et al. Phenotype of mice and macrophages deficient in both phagocyte oxidase and inducible nitric oxide synthase. Immunity 10, 29–38 (1999)

    CAS  Article  Google Scholar 

  4. 4

    Ganz, T. Oxygen-independent microbicidal mechanisms of phagocytes. Proc. Am. Assoc. Physicians 111, 390–395 (1999)

    CAS  Article  Google Scholar 

  5. 5

    Selsted, M. E. & Ouellette, A. J. Defensins in granules of phagocytic and non-phagocytic cells. Trends Cell Biol. 5, 114–119 (1995)

    CAS  Article  Google Scholar 

  6. 6

    Biggar, W. D. & Sturgess, J. M. Role of lysozyme in the microbicidal activity of rat alveolar macrophages. Infect. Immun. 16, 974–982 (1977)

    CAS  PubMed  PubMed Central  Google Scholar 

  7. 7

    del Cerro-Vadillo, E. et al. Cutting edge: a novel nonoxidative phagosomal mechanism exerted by cathepsin-D controls Listeria monocytogenes intracellular growth. J. Immunol. 176, 1321–1325 (2006)

    Article  Google Scholar 

  8. 8

    Hiemstra, P. S., van den Barselaar, M. T., Roest, M., Nibbering, P. H. & van Furth, R. Ubiquicidin, a novel murine microbicidal protein present in the cytosolic fraction of macrophages. J. Leukoc. Biol. 66, 423–428 (1999)

    CAS  Article  Google Scholar 

  9. 9

    Hiemstra, P. S. et al. Antimicrobial proteins of murine macrophages. Infect. Immun. 61, 3038–3046 (1993)

    CAS  PubMed  PubMed Central  Google Scholar 

  10. 10

    Shipley, J. M., Wesselschmidt, R. L., Kobayashi, D. K., Ley, T. J. & Shapiro, S. D. Metalloelastase is required for macrophage-mediated proteolysis and matrix invasion in mice. Proc. Natl Acad. Sci. USA 93, 3942–3946 (1996)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Hautamaki, R. D., Kobayashi, D. K., Senior, R. M. & Shapiro, S. D. Macrophage elastase is required for cigarette smoke-induced emphysema in mice. Science 277, 2002–2004 (1997)

    CAS  Article  Google Scholar 

  12. 12

    Brinckerhoff, C. E. & Matrisian, L. M. Matrix metalloproteinases: a tail of a frog that became a prince. Nature Rev. Mol. Cell Biol. 3, 207–214 (2002)

    CAS  Article  Google Scholar 

  13. 13

    Raza, S. L., Nehring, L. C., Shapiro, S. D. & Cornelius, L. A. Proteinase-activated receptor-1 regulation of macrophage elastase (MMP-12) secretion by serine proteinases. J. Biol. Chem. 275, 41243–41250 (2000)

    CAS  Article  Google Scholar 

  14. 14

    Lijnen, H. R. et al. Temporal and topographic matrix metalloproteinase expression after vascular injury in mice. Thromb. Haemost. 81, 799–807 (1999)

    CAS  Article  Google Scholar 

  15. 15

    Knauper, V. et al. The role of the C-terminal domain of human collagenase-3 (MMP-13) in the activation of procollagenase-3, substrate specificity, and tissue inhibitor of metalloproteinase interaction. J. Biol. Chem. 272, 7608–7616 (1997)

    CAS  Article  Google Scholar 

  16. 16

    Tam, E. M., Moore, T. R., Butler, G. S. & Overall, C. M. Characterization of the distinct collagen binding, helicase and cleavage mechanisms of matrix metalloproteinase 2 and 14 (gelatinase A and MT1-MMP): the differential roles of the MMP hemopexin c domains and the MMP-2 fibronectin type II modules in collagen triple helicase activities. J. Biol. Chem. 279, 43336–43344 (2004)

    CAS  Article  Google Scholar 

  17. 17

    McQuibban, G. A. et al. Matrix metalloproteinase activity inactivates the CXC chemokine stromal cell-derived factor-1. J. Biol. Chem. 276, 43503–43508 (2001)

    CAS  Article  Google Scholar 

  18. 18

    McQuibban, G. A. et al. Inflammation dampened by gelatinase A cleavage of monocyte chemoattractant protein-3. Science 289, 1202–1206 (2000)

    ADS  CAS  Article  Google Scholar 

  19. 19

    Gronski, T. J. et al. Hydrolysis of a broad spectrum of extracellular matrix proteins by human macrophage elastase. J. Biol. Chem. 272, 12189–12194 (1997)

    CAS  Article  Google Scholar 

  20. 20

    Belaaouaj, A. et al. Mice lacking neutrophil elastase reveal impaired host defense against gram-negative bacterial sepsis. Nature Med. 4, 615–618 (1998)

    CAS  Article  Google Scholar 

  21. 21

    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 

  22. 22

    Weinrauch, Y., Drujan, D., Shapiro, S., Weiss, J. & Zychlinsky, A. Neutrophil elastase targets virulence factors of enterobacteria. Nature 417, 91–94 (2002)

    ADS  CAS  Article  Google Scholar 

  23. 23

    Lehrer, R. I. & Ganz, T. Antimicrobial polypeptides of human neutrophils. Blood 76, 2169–2181 (1990)

    CAS  PubMed  Google Scholar 

  24. 24

    Wilson, C. L. et al. Regulation of intestinal alpha-defensin activation by the metalloproteinase matrilysin in innate host defense. Science 286, 113–117 (1999)

    CAS  Article  Google Scholar 

  25. 25

    Vu, T. H. et al. MMP-9/gelatinase B is a key regulator of growth plate angiogenesis and apoptosis of hypertrophic chondrocytes. Cell 93, 411–422 (1998)

    CAS  Article  Google Scholar 

  26. 26

    Li, Q., Park, P., Wilson, C. & Parks, W. Matrilysin shedding of syndecan-1 regulates chemokine mobilization and transepithelial efflux of neutrophils in acute lung injury. Cell 111, 635–646 (2002)

    CAS  Article  Google Scholar 

  27. 27

    Curci, J. A., Liao, S., Huffman, M. D., Shapiro, S. D. & Thompson, R. W. Expression and localization of macrophage elastase (matrix metalloproteinase-12) in abdominal aortic aneurysms. J. Clin. Invest. 102, 1900–1910 (1998)

    CAS  Article  Google Scholar 

  28. 28

    Heymans, S. et al. Inhibition of plasminogen activators or matrix metalloproteinases prevents cardiac rupture by impairs therapeutic angiogenesis and causes cardiac failure. Nature Med. 5, 1135–1142 (1999)

    CAS  Article  Google Scholar 

  29. 29

    Golubkov, V. S. et al. Membrane type-1 matrix metalloproteinase (MT1-MMP- exhibits an important intracellular cleavage function and causes chromosome instability. J. Biol. Chem. 280, 25079–25086 (2005)

    CAS  Article  Google Scholar 

  30. 30

    Limb, G. A. et al. Matrix metalloprteinase-1 associates with intracellular organelles and confers resistance to laminin a/c degradation during apoptosis. Am. J. Pathol. 166, 1555–1563 (2005)

    CAS  Article  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the National Institutes of Health (NIH) National Heart, Lung and Blood Institute (NHLBI) (S.D.S.) and Spanish and European public agencies (F.X.G.-R.).

Author Contributions A.M.H. performed in vivo and in vitro studies, contributed to data interpretation and mechanistic advance, prepared the manuscript and figures, and performed all revisions. W.O.H. performed in vivo and in vitro studies and contributed to study design, data analysis, and mechanistic advance. C.S.R. performed CTD processing studies. F.X.G.-R. constructed the three-dimensional homology model of MMP12 CTD. S.D.S. generated Mmp12-/- mice and all recombinant proteins, was responsible for study design, data interpretation, and mechanistic advance, and assisted with manuscript preparation.

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Correspondence to A. McGarry Houghton.

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Houghton, A., Hartzell, W., Robbins, C. et al. Macrophage elastase kills bacteria within murine macrophages. Nature 460, 637–641 (2009). https://doi.org/10.1038/nature08181

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