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

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.

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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.

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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.

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Correspondence to Charles L. Bevins.

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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

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