Molecular characterization of the involvement of phospholipase A2 (PLA2) in the immune response to mastitis pathogens in dairy cattle in vivo and in vitro

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Zhongde Wang


Mastitis is the single most costly disease of dairy cattle, causing substantial losses through reduction of milk production, milk wastage, treatment costs and loss of animals. Development of more effective treatments than are currently available for bovine mastitis is needed. The mammalian enzyme phospholipase A2 (PLA2) has been shown in a mouse model of mastitis to reduce mammary gland inflammation and damage after bacterial induced mastitis, yet the importance of PLA2 in the bovine response to mastitis remain unknown. This study will examine PLA2 expression by the bovine mammary gland in vivo as a result of mastitis, as well as the effects of exogenously administered PLA2 on bovine mammary epithelial cells in vitro. Expression levels of PLA2 and cytokines IL-1alpha, IL-1beta, IL-6, IL-8, IL-10, TNF-alpha and CCL5 will be measured by qRTPCR and compared between mastitic and unaffected mammary gland quarters within individual cows. Additionally, expression of these cytokines and PLA2 by bovine mammary epithelial cells in vitro will be compared before and after challenge by lipopolysaccharide from Escherichia coli, with and without concurrent exposure to exogenous PLA2. We hypothesize that in vivo and in vitro expression of PLA2 will vary consistently depending on infection status of mammary gland tissue and in vitro cultures, and that exogenously administered PLA2 will result in a reduced inflammatory cytokine profile in cultured bovine mammary epithelial cells. Results will be used to determine whether PLA2 should be further pursued as a potential therapeutic agent for the treatment of bovine mastitis.

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