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TNFα-mediated plasminogen activation on neutrophils is involved in the high plasmin activity in mammary secretion of drying-off cows

Published online by Cambridge University Press:  29 July 2009

Wen K Chou
Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan ROC
Ting C Yu
Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan ROC
Shuen E Chen
Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan ROC
Ho C Peh
Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan ROC
Wen B Liu
Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan ROC
Ming T Chen
Department of Bioindustry Technology, Da Yeh University, Chung Hwa 555, Taiwan ROC
Hajime Nagahata
Department of Animal Health, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501, Japan
Chai J Chang*
Department of Animal Science, National Chung Hsing University, Taichung 402, Taiwan ROC
*For correspondence; e-mail:


Interactions between inflammatory cytokines and plasminogen (Pg) activation system on immune cells are yet to be established. In previous studies we reported a somatic cell-associated elevation of proteolytic activity in mammary secretion of drying-off goats and cows. The purposes of the present study were to examine the role of TNF-α in polymorphonuclear neutrophil (PMN)-associated Pg activation, and the significance of this activation pathway for overall plasmin (Pm) activity in mammary secretion of drying-off cows. Results of experiments in vitro showed that the spontaneous Pg activation observed on fresh preparations of bovine blood PMN was completely blocked by anti bovine TNF-α antibody, and was further up-regulated by exogenous bovine TNF-α. Monitoring the parameters of mammary secretion of drying-off cows revealed that both somatic cell counts and differential PMN ratio was significantly elevated at weeks 1, 2 and 3 of milk stasis. Nevertheless, specific activity of soluble Pm in mammary secretion increased and the level of 17-kDa TNF-α decreased immediately following milk stasis. Iimmunoblotting revealed that although both 26-kDa pro-TNF-α and 17-kDa TNF-α were consistently present in somatic cells of mammary secretion collected at weeks 0, 1, 2 and 3 of milk stasis, only 26-kDa pro-TNF-α was present in somatic cells of milk during lactation. In-vitro assay indicated that cell-free mammary secretion of drying-off cows exerted no Pg activation bioactivity towards bovine blood PMN. Altogether, the current study suggests the existence of an active TNF-α-Pg-Pm autocrine/paracrine loop on the massively infiltrated PMN inside udders of drying-off cows, which involves extensive binding and internalization of 17-kDa TNF-α on PMN and consequently activation of Pg, resulting in high Pm activity and low 17-kDa TNF-α level in mammary secretion. These coordinated mechanisms may play a role in the defence of drying-off mammary gland.

Research Article
Copyright © Proprietors of Journal of Dairy Research 2009

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