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Prolactin-Induced Activation of Nuclear Factor B in Bovine Mammary Epithelial Cells: Role in Chronic Mastitis

P. Boutet^*,1, J. Sulon^*, R. Closset^*, J. Detilleux, J.-F. Beckers^*, F. Bureau^* and P. Lekeux^*

^* Department for Functional Sciences, and
Department of Animal Production, Faculty of Veterinary Medicine, University of Liège, B-4000 Liège, Belgium

¹ Corresponding author: philippe.boutet{at}ulg.ac.be

We sought to determine whether prolactin (PRL) could influence the neutrophilic inflammation that characterizes chronic mastitis. Most of the genes encoding inflammatory proteins depend on the nuclear factor B (NF-B) for their expression. We addressed the hypothesis that immunomodulatory activities of PRL might arise from an increase in NF-B activity. MAC-T cells, a bovine mammary epithelial cell line, were stimulated with increasing concentrations of bovine PRL (1, 5, 25, 125, and 1,000 ng/mL). Level of NF-B binding activity was measured and mRNA was evaluated for IL-1ß, IL-6, IL-8, granulocyte-macrophage colony-stimulating factor (GMCSF), IFN-, and tumor necrosis factor (TNF)-, cytokines known to require NF-B for their maximal transcription. Prolactin activated NF-B; maximal NF-B activation was weaker with PRL than with TNF- at 30 or 180 min poststimulation. In addition, PRL significantly amplified, in a dose-dependent manner, mRNA expression of IL-1ß, IL-6, IL-8, GMCSF, and TNF-. We measured PRL concentrations in blood and milk from healthy and chronic mastitis-infected cows, and studied the relationship between the PRL concentration and the degree of inflammation in the mammary gland as indirectly assessed by somatic cell counts (SCC). Plasma PRL did not differ significantly between healthy and chronic mastitis-affected cows (63.7 and 67.5 ng/mL, respectively). Milk PRL concentration was significantly increased in chronic mastitis-affected quarters with the highest SCC, and had a positive significant correlation between SCC, as well as between the number of neutrophils present in milk samples. The present findings show that PRL promotes an inflammatory response in bovine mammary epithelial cells via NF-B activation, and suggest a role for PRL in the pathogenesis of chronic mastitis.

Key Words: bovine mastitis • prolactin • nuclear factor B • cytokine