Effect of Lipopolysaccharide and Muramyl Dipeptide on Apoptosis of Bovine Mammary Gland Lymphocytes

SIMPLE SUMMARY: Inflammation of the mammary gland in dairy cattle is a global problem and causes huge financial loss to dairy farmers. Inflammation is caused by many species of bacteria penetrating through the teat canals into the udder. Those bacteria are usually eliminated by treatment with intramammary injection of antibiotics, while they are also eliminated by the immune cells of the cow. One of the immune cells are lymphocytes which are responsible for specific immunity. When viable, they are able to carry out their normal functions. The present study focused on the investigation of cell death of lymphocytes during bovine mammary gland inflammation. We analyzed apoptosis in mammary gland lymphocytes under the stimulation of lipopolysaccharides and muramyl dipeptide as the endotoxin of Gram-negative bacteria and the natural content of the cell wall of Gram-positive bacteria. We found that they induce lymphocyte apoptosis in the early phase of inflammation, which can be associated with the expression of CD44 receptors on lymphocytes. This receptor is important in many physiological processes, including apoptosis of cells. For a better understanding of immune responses in mammary glands and for developing of immunotherapy without antibiotics, the process of inflammation, including cell death of immune cells necessitates further holistic studies. ABSTRACT: The aim of this study was to evaluate whether apoptosis of lymphocytes is modulated by stimulation by lipopolysaccharide (LPS) of Escherichia coli or muramyl dipeptide (MDP). Cell populations were obtained by lavaging of the mammary glands 24, 48, 72, and 168 h following intramammary induced inflammation. The portion of apoptotic lymphocytes peaked at 48 h after treatment with LPS or MDP. The analysis of CD44 expression of the same cell populations showed a higher percentage of CD44-positive lymphocytes 24- and 48-h following induction of inflammation by LPS or MDP. The results demonstrate that during both experimental infection of bovine mammary glands with LPS or MDP, apoptosis of lymphocytes was induced in the initial phase of the inflammatory response and CD44 was also overexpressed at the beginning of inflammation. These data suggest a connection of lymphocyte apoptosis with the expression of CD44 receptors.