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* Immunology and Disease Resistance Laboratory and
Veterinary Services, USDA-Agricultural Research Service, Beltsville, MD 20705
Department of Animal and Avian Sciences, University of Maryland, College Park 20742
Corresponding author: D. D. Bannerman; e-mail: dbanner{at}anri.barc.usda.gov.
Several species of gram-negative bacteria, including Escherichia coli, Klebsiella pneumoniae, and various species of Enterobacter, are common mastitis pathogens. All of these bacteria are characterized by the presence of endotoxin or lipopolysaccharide (LPS) in their outer membrane. The bovine mammary gland is highly sensitive to LPS, and LPS has been implicated, in part, in the pathogenesis of gram-negative mastitis. Recognition of LPS is a key event in the innate immune response to gram-negative infection and is mediated by the accessory molecules CD14 and LPS-binding protein (LBP). The objective of the current study was to determine whether LBP levels increased in the blood and mammary gland following LPS challenge. The left and right quarters of five midlactating Holstein cows were challenged with either saline or LPS (100 µg), respectively, and milk and blood samples collected. Basal levels of plasma and milk LBP were 38 and 6 µg/ml, respectively. Plasma LBP levels increased as early as 8 h post-LPS challenge and reached maximal levels of 138 µg/ml by 24 h. Analysis of whey samples derived from LPS-treated quarters revealed an increase in milk LBP by 12 h. Similar to plasma, maximal levels of milk LBP (34 µg/ml) were detected 24 h following the initial LPS challenge. Increments in milk LBP levels paralleled a rise in soluble CD14 (sCD14) levels and initial rises in the levels of these proteins were temporally coincident with maximal neutrophil recruitment to the inflamed gland. Because LBP and sCD14 are known to enhance LPS-induced host cell activation and to facilitate detoxification of LPS, these data are consistent with a role for these molecules in mediating mammary gland responses to LPS.
Key Words: lipopolysaccharide-binding protein • soluble CD14 • endotoxin • mastitis
Abbreviation key: HRP = horseradish peroxidase, LBP = LPS-binding protein, mCD14 = membrane-bound CD14, MSCC = milk somatic cell count, sCD14 = soluble CD14, TBS = Tris-buffered saline, Tlr = Toll-like receptor
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