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Elevated Milk Soluble CD14 in Bovine Mammary Glands Challenged with Escherichia coli Lipopolysaccharide

J.-W. Lee^*, M. J. Paape, T. H. Elsasser and X. Zhao^*

^* Department of Animal Science, McGill University, Ste-Anne-de-Bellevue, Quebec, H9X 3V9, Canada; and
Immunology and Disease, Resistance Laboratory and
Growth Biology Laboratory, USDA-ARS, Beltsville, MD 20705

Corresponding author:
X. Zhao; e-mail:
Zhao{at}macdonald.mcgill.ca.

The purpose of this study was to determine whether soluble CD14 (sCD14) in milk was affected by stage of lactation, milk somatic cell count (SCC), presence of bacteria, or lipopolysaccharide (LPS)-induced inflammation. Milk samples from 100 lactating cows (396 functional quarters) were assayed for sCD14 in milk to determine effects of stage of lactation, SCC, and intramammary infection. The concentration of sCD14 was highest in transitional milk (0 to 4 d postpartum) and in milk with high SCC (> 750,000 cells/ml). Most of the infected quarters (> 80%) were infected by coagulase-negative staphylococci and yeast. No difference was found between noninfected and infected quarters. One quarter of six healthy lactating cows was challenged with 100 µg LPS in order to study the kinetics of sCD14 during an LPS-induced inflammation. Milk samples were collected at various intervals until 72 h after injection. Rectal temperature, milk tumor necrosis factor-, and interleukin-8 increased immediately after challenge. The increase in sCD14 paralleled the increase in SCC, peaked at 12 h, and started to decline after 24 h. Serum leakage, as characterized by the level of bovine serum albumin in milk, peaked at 4 h and then gradually decreased. All parameters remained at basal levels in control quarters throughout the study. In vitro experiments indicated that neutrophils released sCD14 in response to LPS in a dose-dependent manner. The results indicate that the concentration of sCD14 was significantly increased in milk after LPS challenge. The increase was not likely due to serum leakage. Instead, infiltrated neutrophils might be the main source of increased sCD14 in milk during inflammation.

Key Words: CD14 • LPS • Escherichia coli • mastitis

Abbreviation key: BAL = bronchoalveolar lavage, BSA = bovine serum albumin, CSF = cerebrospinal fluid, IL = interleukin, LBP = lipopolysaccharide-binding protein, LPS = lipopolysaccharide, TNF = tumor necrosis factor

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