Article

Induction of Nitric Oxide Production Mediated by Tumor Necrosis Factor Alpha on Staphylococcal Enterotoxin C-Stimulated Bovine Mammary Gland Cells

T-Cell Research Institute, Minami-Yoshinari, Aobaku, Japan.
Clinical and Diagnostic Laboratory Immunology (Impact Factor: 2.51). 02/2004; 11(1):203-10. DOI: 10.1128/CDLI.11.1.203-210.2004
Source: PubMed

ABSTRACT

Mammary gland (MG) secretions (MGS) derived from secretory cows infected with coagulase-negative staphylococci (CoNS) showed somatic cell counts and lactoferrin similar to levels found in the MGS of secretory cows infected with Staphylococcus aureus. However, nitrite and nitrate (NOx) and staphylococcal enterotoxin C (SEC) were found in MGS infected with S. aureus at much higher levels than in cows infected with CoNS. These results suggested that NOx could be intimately correlated with the production of SEC in secretory cows infected with S. aureus. Therefore, we examined the production of NOx and the expression of proinflammatory cytokines and microsomal cytochrome P450 (CYP450) after injection of SEC into the MGS of secretory cows. We were able to detect NOx and the proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) on MG cells of SEC-injected MGS. It was also found that CYP450 in the MG cells from SEC-injected MGS was down-regulated by approximately one-third in comparison with the cells from phosphate-buffered saline-injected MGS. This in vitro system also showed that NOx could be induced in the culture of bovine macrophage-lined cells (FBM-17) with the supernatants of SEC-stimulated bovine peripheral blood lymphocytes (BoPBLs) but not in the culture of peripheral mononuclear cells with SEC-stimulated BoPBLs. The expression of the mRNA for both inducible nitric oxide synthase and TNF-alpha in FBM-17 was enhanced by culturing with the supernatant of SEC-stimulated BoPBLs, although CYP450 was down-regulated. These results indicate that the down-regulation of CYP450 was caused by the production of TNF-alpha in SEC-stimulating MG cells containing macrophages and via NOx production. Therefore, we suggest that NOx released from activated MG cells via the superantigenic activity of SEC caused oxidative damage to the MG in S. aureus-induced mastitis.

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    • "NOx is a free radical that mediates physiological and pathological changes in the body, is synthesized from L-arginine by inducible nitric oxide synthase (iNOS; Archer, 1993) and mediated from endothelial, epithelial, neuronal and phagocytic cells (Palmer et al., 1987; Onado and Inano, 1998). However, NOx becomes a cytotoxic mediator when it is released in excessive amounts during Escherichia coli (E. coli ) infections and in response to intramammary injection of lipopolysaccharide (LPS; Boulanger et al., 2001; Burvenich et al., 2003; Komine et al., 2004). The objective of this study was to detect NOx concentrations in mammary secretions from infected and non-infected mammary quarters of heifers before and after calving, and to evaluate the relationship of NOx with bacterial species and somatic cell count (SCC). "
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