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Lysostaphin expression in mammary glands confers protection against staphylococcal infection in transgenic mice

Nature Biotechnology volume 19, pages 66–70 (2001)Cite this article

Abstract

Infection of the mammary gland, in addition to causing animal distress, is a major economic burden of the dairy industry. Staphylococcus aureus is the major contagious mastitis pathogen, accounting for approximately 15–30% of infections, and has proved difficult to control using standard management practices. As a first step toward enhancing mastitis resistance of dairy animals, we report the generation of transgenic mice that secrete a potent anti-staphylococcal protein into milk. The protein, lysostaphin, is a peptidoglycan hydrolase normally produced by Staphylococcus simulans. When the native form is secreted by transfected eukaryotic cells it becomes glycosylated and inactive. However, removal of two glycosylation motifs through engineering asparagine to glutamine codon substitutions enables secretion of Gln125,232-lysostaphin, a bioactive variant. Three lines of transgenic mice, in which the 5′-flanking region of the ovine β-lactoglobulin gene directed the secretion of Gln125,232-lysostaphin into milk, exhibit substantial resistance to an intramammary challenge of 104 colony-forming units (c.f.u.) of S. aureus, with the highest expressing line being completely resistant. Milk protein content and profiles of transgenic and nontransgenic mice are similar. These results clearly demonstrate the potential of genetic engineering to combat the most prevalent disease of dairy cattle.

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Figure 1: Western blot analysis of lysostaphin production by transfected COS-7 cells.
Figure 2: Bacterial plate assay for staphylolytic activity.
Figure 3: Proteins in milk collected from mice on day 10 of lactation.
Figure 4: Western blot analysis of lysostaphin in milk collected from mice on day 10 of lactation.
Figure 5: Hematoxylin and eosin stained sections of lactating mouse mammary tissue obtained 24 h after an intramammary infusion of S. aureus (104 c.f.u.).

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Acknowledgements

We thank Juli Foster, Leah Schulman, Barbara Hughes, and Francis Kinghorn for excellent technical assistance. The work was funded in part by grants from The Northeast Dairy Foods Research Center, the Vermont Dairy Promotion Council, and the New England Dairy Promotion Board.

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Author notes

  1. Christine M. Williamson

    Present address: MRC Mammalian Genetics Unit, Harwell, Didcot, Oxfordshire, OX11 0RD, UK

  2. Alistair J. Lax

    Present address: Oral Microbiology, King's College London, London, SE1 9RT, UK

  3. Karen Moore

    Present address: Department of Animal Sciences, University of Florida, Gainesville, FL, 32611

Authors and Affiliations

  1. Department of Animal Sciences, University of Vermont, Burlington, 05405, VT

    David E. Kerr, Karen Plaut & A. John Bramley

  2. Institute for Animal Health, Compton Laboratory, Berks, RG20 7NN, UK

    A. John Bramley, Christine M. Williamson & Alistair J. Lax

  3. United States Department of Agriculture, Gene Evaluation and Mapping Laboratory, Agricultural Research Service, Beltsville, 20705, MD

    Karen Moore, Kevin D. Wells & Robert J. Wall

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  1. David E. Kerr
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Correspondence to David E. Kerr.

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Kerr, D., Plaut, K., Bramley, A. et al. Lysostaphin expression in mammary glands confers protection against staphylococcal infection in transgenic mice. Nat Biotechnol 19, 66–70 (2001). https://doi.org/10.1038/83540

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