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Infection and Immunity, November 2006, p. 6206-6212, Vol. 74, No. 11
0019-9567/06/$08.00+0     doi:10.1128/IAI.00238-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Transgenic Cows That Produce Recombinant Human Lactoferrin in Milk Are Not Protected from Experimental Escherichia coli Intramammary Infection{triangledown}

P. Hyvönen,1* L. Suojala,2 T. Orro,2 J. Haaranen,1 O. Simola,2 C. Røntved,3 and S. Pyörälä2

Institute of Applied Biotechnology, University of Kuopio, P.O. Box 1627, FI-70211 Kuopio, Finland,1 Department of Clinical Veterinary Science, Saari Unit, Faculty of Veterinary Medicine, University of Helsinki, FI-04920 Saarentaus, Finland,2 Department of Animal Health, Welfare and Nutrition, Research Centre Foulum, Danish Institute of Agricultural Science, DK-8830 Tjele, Denmark3

Received 13 February 2006/ Returned for modification 23 March 2006/ Accepted 8 August 2006

This is the first study describing an experimental mastitis model using transgenic cows expressing recombinant human lactoferrin (rhLf) in their milk. The aim of the study was to investigate the concentrations in milk and protective effects of bovine and recombinant human lactoferrin in experimental Escherichia coli mastitis. Experimental intramammary infection was induced in one udder quarter of seven first-lactating rhLf-transgenic cows and six normal cows, using an E. coli strain isolated from cows with clinical mastitis and known to be susceptible to Lf in vitro. Clinical signs were recorded during the experimental period, concentrations of human and bovine Lf and indicators of inflammation and bacterial counts were determined for milk, and concentrations of acute-phase proteins and tumor necrosis factor alpha were determined for sera and milk. Serum cortisol and blood hematological and biochemical parameters were also determined. Expression levels of rhLf in the milk of transgenic cows remained constant throughout the experiment (mean, 2.9 mg/ml). The high Lf concentrations in the milk of transgenic cows did not protect them from intramammary infection. All cows became infected and developed clinical mastitis. The rhLf-transgenic cows showed milder systemic signs and lower serum cortisol and haptoglobin concentrations than did controls. This may be explained by lipopolysaccharide-neutralizing and immunomodulatory effects of the high Lf concentrations in their milk. However, Lf does not seem to be a very efficient protein for genetic engineering to enhance the mastitis resistance of dairy cows.

* Corresponding author. Mailing address: Institute of Applied Biotechnology, University of Kuopio, P.O. Box 1627, FI-70211 Kuopio, Finland. Phone: 358 17 163564. Fax: 358 17 163322. E-mail: paula.hyvonen{at}uku.fi.

{triangledown} Published ahead of print on 5 September 2006.

Editor: J. L. Flynn

Infection and Immunity, November 2006, p. 6206-6212, Vol. 74, No. 11
0019-9567/06/$08.00+0     doi:10.1128/IAI.00238-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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