Activation of phagocytes during initiation and resolution of mammary gland injury induced by lipopolysaccharide in heifers

The object of the study was the comparative assessment of phagocyte activation during initiation and resolution of mammary gland injury induced by lipopolysaccharide (LPS) or buffered salt solution (PBS) on the basis of the CD14 receptor positivity. The experiments were carried out in 15 clinically normal Holstein x Bohemian Red Pied crossbred heifers, aged 14 to 18 months. Noninflammatory and inflammatory mammary gland injury were induced by intramammary administration of PBS (10 mL) and LPS (10 mL, 1 microg/mL), respectively. Samples of the cell populations were obtained by mammary lavages at 24 h intervals. Flow cytometry was used to determine the CD14+ neutrophils, monocytes, and macrophages. The percentage of CD14+ neutrophils was only 1.2% and 1.3% 24 h after the treatment with PBS and LPS, respectively. The resolution was accompanied by an increase in proportion of CD14+ neutrophils. The proportion of CD14+ neutrophils returned to initial values in the PBS-treated, but not in the LPS-treated mammary glands till 96 h. Percentage of CD14+ monocytes increased after 24 h and the effect was more pronounced in the LPS-treated than in the PBS treated mammary glands (P < 0.05). The percentage of CD14+ macrophages decreased highly significantly at 24 h in the LPS-treated, but not in the PBS-treated mammary glands (P < 0.01). The resolution of mammary gland injury (48 to 96 h) was characterised by an increase in CD14+ macrophages proportion, which was greater in the LPS-treated than PBS-treated mammary glands (P < 0.01). The activation of macrophages during resolution of mammary gland injury can be interpreted as an important mechanism of restitution.