Dysfunction of neutrophil during parturition and proteome analysis of placenta and serum in cow

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The purpose of this project is to identify and characterize internal factors to reduce neutrophil functions after parturion in cows, since bovine mastitis sometimes occurs during parturition. In the experiments, the analysis of signal transduction pathways for activation of bovine neutrophils was performed. As important molecules for activation of NADPH oxidase in bovine neutrophils, cPKC, p38MAPK, ERK, PI3K and nPKC(pkcδ) were identified, but these factors did not changes during parturition in the cow. Furthermore, I tried to identify causative factors to reduce immune functions of cow after parturition by using two dimensional electrophoresis and other proteome technique. The proteome analysis showed that lactoferrin, transferrin, bovine pregnancy related glycoprotein (bPAG) and keton body in serum and placenta playd as suppressive factors for NADPH oxidase activation of bovine neutrophils. Moreover, to clarify the mechanism of this suppressive effects on neutrophil function, the effects of lactoferrin on LPS-or conA-induced expression of mRNA of INF-gamma, IL-1beta, TNF-alpha, IL12p4O and iNOS in mouse macrophage and feline monocyte was examined. The results indicated that the treatment of lactoferrin decreased LPS-or conA-induced expression of mRNA of INF-gamma and IL-1beta but did not affect that of TNF-alpha, IL12p4O and iNOS. The addition of lactoferrin to culture medium was showed to reduce differentiation form myeloblast to neutrophil in HL60 cells. These results suggest that lactoferrin was associated with regulation of inflammation and immune system mammary gland.