Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Protection against enteric salmonellosis in transgenic mice expressing a human intestinal defensin

Nature volume 422, pages 522–526 (2003)Cite this article

Abstract

Genetically encoded antibiotic peptides are evolutionarily ancient and widespread effector molecules of immune defence1,2,3. Mammalian defensins, one subset of such peptides, have been implicated in the antimicrobial defence capacity of phagocytic leukocytes and various epithelial cells4, but direct evidence of the magnitude of their in vivo effects have not been clearly demonstrated. Paneth cells, specialized epithelia of the small intestinal crypt, secrete abundant α-defensins and other antimicrobial polypeptides5,6 including human defensin 5 (HD-5; also known as DEFA5)7,8,9. Although antibiotic activity of HD-5 has been demonstrated in vitro9,10, functional studies of HD-5 biology have been limited by the lack of in vivo models. To study the in vivo role of HD-5, we developed a transgenic mouse model using a 2.9-kilobase HD-5 minigene containing two HD-5 exons and 1.4 kilobases of 5′-flanking sequence. Here we show that HD-5 expression in these mice is specific to Paneth cells and reflects endogenous enteric defensin gene expression. The storage and processing of transgenic HD-5 also matches that observed in humans. HD-5 transgenic mice were markedly resistant to oral challenge with virulent Salmonella typhimurium. These findings provide support for a critical in vivo role of epithelial-derived defensins in mammalian host defence.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Germline transmission and characterization of HD-5 transgene expression in mice.
Figure 2: Western blot analysis of ileal HD-5 peptide.
Figure 3: Challenge of HD-5 transgenic and wild-type FvB control mice with virulent S. typhimurium.

Similar content being viewed by others

References

  1. Boman, H. G. Peptide antibiotics and their role in innate immunity. Annu. Rev. Immunol. 13, 61–92 (1995)

    Article  CAS  Google Scholar 

  2. Meister, M., Hetru, C. & Hoffmann, J. A. The antimicrobial host defense of Drosophila. Curr. Top. Microbiol. Immunol. 248, 17–36 (2000)

    CAS  Google Scholar 

  3. Zasloff, M. Antimicrobial peptides of multicellular organisms. Nature 415, 389–395 (2002)

    Article  ADS  CAS  Google Scholar 

  4. Lehrer, R. I., Bevins, C. L. & Ganz, T. in Mucosal Immunology (eds Ogra, P. L., Mestecky, J., Lamm, M. E., Strober, W. M. & Bienstock, J.) 89–99 (Academic, New York, 1998)

    Google Scholar 

  5. Ayabe, T. et al. Secretion of microbicidal α-defensins by intestinal Paneth cells in response to bacteria. Nature Immunol. 1, 113–118 (2000)

    Article  CAS  Google Scholar 

  6. Porter, E. M., Bevins, C. L., Ghosh, D. & Ganz, T. The multifaceted Paneth cell. Cell. Mol. Life Sci. 59, 156–170 (2002)

    Article  CAS  Google Scholar 

  7. Jones, D. E. & Bevins, C. L. Paneth cells of the human small intestine express an antimicrobial peptide gene. J. Biol. Chem. 267, 23216–23225 (1992)

    CAS  PubMed  Google Scholar 

  8. Porter, E., Liu, L., Oren, A., Anton, P. & Ganz, T. Localization of human intestinal defensin 5 in Paneth cell granules. Infect. Immun. 65, 2389–2395 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Ghosh, D. et al. Paneth cell trypsin is the processing enzyme for human defensin-5. Nature Immunol. 3, 583–590 (2002)

    Article  CAS  Google Scholar 

  10. Porter, E., van Dam, E., Valore, E. & Ganz, T. Broad spectrum antimicrobial activity of human intestinal defensin 5. Infect. Immun. 65, 2396–2401 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Ouellette, A. J. & Bevins, C. L. in Development of the Gastrointestinal Tract (eds Sanderson, I. R. & Walker, W. A.) 147–164 (Decker, Hamilton, Ontario, 1999)

    Google Scholar 

  12. Mallow, E. B. et al. Human enteric defensins: gene structure and developmental expression. J. Biol. Chem. 271, 4038–4045 (1996)

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  14. Darmoul, D. & Ouellette, A. J. Positional specificity of defensin gene expression reveals Paneth cell heterogeneity in mouse small intestine. Am. J. Physiol. 271, G68–G74 (1996)

    CAS  PubMed  Google Scholar 

  15. Wang, M. S., Pang, J. S. & Selsted, M. E. Semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels. Anal. Biochem. 253, 225–230 (1997)

    Article  CAS  Google Scholar 

  16. Cole, A. M. & Ganz, T. Human antimicrobial peptides: analysis and application. Biotechniques 29, 822–831 (2000)

    Article  CAS  Google Scholar 

  17. Porter, E. M. et al. Isolation of human intestinal defensins from ileal neobladder urine. FEBS Lett. 434, 272–276 (1998)

    Article  CAS  Google Scholar 

  18. Cunliffe, R. N. et al. Human defensin 5 is stored in precursor form in normal Paneth cells and is expressed by some villous epithelial cells and by metaplastic Paneth cells in the colon in inflammatory bowel disease. Gut 48, 176–185 (2001)

    Article  CAS  Google Scholar 

  19. Satoh, Y., Habara, Y., Ono, K. & Kanno, T. Carbamylcholine- and catecholamine-induced intracellular calcium dynamics of epithelial cells in mouse ileal crypts. Gastroenterology 108, 1345–1356 (1995)

    Article  CAS  Google Scholar 

  20. Ouellette, A. J., Satchell, D. P., Hsieh, M. M., Hagen, S. J. & Selsted, M. E. Characterization of luminal Paneth cell α-defensins in mouse small intestine: attenuated antimicrobial activities of peptides with truncated amino termini. J. Biol. Chem. 275, 33969–33973 (2000)

    Article  CAS  Google Scholar 

  21. Selsted, M. E., Miller, S. I., Henschen, A. H. & Ouellette, A. J. Enteric defensins: antibiotic peptide components of intestine host defense. J. Cell Biol. 118, 929–936 (1992)

    Article  CAS  Google Scholar 

  22. Putsep, K. et al. Germ-free and colonized mice generate the same products from enteric prodefensins. J. Biol. Chem. 275, 40478–40482 (2000)

    Article  CAS  Google Scholar 

  23. Nizet, V. et al. Innate antimicrobial peptide protects the skin from invasive bacterial infection. Nature 414, 454–457 (2001)

    Article  ADS  CAS  Google Scholar 

  24. Couto, M. A., Liu, L., Lehrer, R. I. & Ganz, T. Inhibition of intracellular Histoplasma capsulatum replication by murine macrophages that produce human defensin. Infect. Immun. 62, 2375–2378 (1994)

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Bals, R., Weiner, D. J., Moscioni, A. D., Meegalla, R. L. & Wilson, J. M. Augmentation of innate host defense by expression of a cathelicidin antimicrobial peptide. Infect. Immun. 67, 6084–6089 (1999)

    CAS  PubMed  PubMed Central  Google Scholar 

  26. Kisich, K. O., Heifets, L., Higgins, M. & Diamond, G. Antimycobacterial agent based on mRNA encoding human β-defensin 2 enables primary macrophages to restrict growth of Mycobacterium tuberculosis. Infect. Immun. 69, 2692–2699 (2001)

    Article  CAS  Google Scholar 

  27. Taketo, M. et al. FVB/N: an inbred mouse strain preferable for transgenic analyses. Proc. Natl Acad. Sci. USA 88, 2065–2069 (1991)

    Article  ADS  CAS  Google Scholar 

  28. Harper, M. E., Barrera-Saldana, H. A. & Saunders, G. F. Chromosomal localization of the human placental lactogen-growth hormone gene cluster to 17q22-24. Am. J. Hum. Genet. 34, 227–234 (1982)

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Bevins, C. L., Jones, D. E., Dutra, A., Schaffzin, J. & Muenke, M. M. Human enteric defensin genes: chromosomal map position and a model of possible evolutionary relationships. Genomics 31, 95–106 (1996)

    Article  CAS  Google Scholar 

  30. Iwabuchi, Y., Katagiri, M. & Masuhara, T. Salivary secretion and histopathological effects after single administration of the muscarinic agonist SNI-2011 in MRL/lpr mice. Arch. Int. Pharmacodyn. Ther. 328, 315–325 (1994)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We acknowledge the technical assistance of J. Russell, S. Lee, D. Wilk, H. deJong and K. Carver. We thank J. Crabb and S. Yadav for their help with peptide analysis, and T. Ganz and E. Porter for discussions and for providing us with recombinant peptides and the polyclonal anti-HD-5 antibody. We also acknowledge discussions with S. Miller and A. Ouellette. This work was supported by grants from the National Institutes of Health (to C.L.B. and N.H.S.) and The Crohn's and Colitis Foundation of America (to N.H.S.). A portion of this work was completed while the indicated authors were at the Departments of Microbiology (N.H.S.) and Pediatrics (N.H.S., C.L.B.), University of Pennsylvania, School of Medicine and the Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.

Author information

Author notes

  1. Nita H. Salzman, Dipankar Ghosh and Yvonne Paterson: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Pediatrics, Division of Gastroenterology, Medical College of Wisconsin, Milwaukee, Wisconsin, 53226, USA

    Nita H. Salzman

  2. Department of Immunology, Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio, 44195, USA

    Dipankar Ghosh & Charles L. Bevins

  3. Division of Neonatology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, 02114, USA

    Kenneth M. Huttner

  4. Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104, USA

    Yvonne Paterson

Authors
  1. Nita H. Salzman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dipankar Ghosh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kenneth M. Huttner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yvonne Paterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Charles L. Bevins
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Charles L. Bevins.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Salzman, N., Ghosh, D., Huttner, K. et al. Protection against enteric salmonellosis in transgenic mice expressing a human intestinal defensin. Nature 422, 522–526 (2003). https://doi.org/10.1038/nature01520

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature01520

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing