Exosomes Derived from Bovine Mammary Epithelial Cells Treated with Transforming Growth Factor-β1 Inhibit the Proliferation of Bovine Macrophages

Transforming growth factor (TGF)-beta 1 is a multifunctional cytokine that plays an important role in regulating immune cell proliferation. We speculate that high expression of TGF-beta 1 may affect the immunity of dairy cows. In this study, untreated exosomes (un-exo) derived from an untreated bovine mammary epithelial cell line (MAC-T) and TGF-beta 1-treated exosomes (t-exo) derived from TGF-beta 1-treated MAC-T cells were isolated by ultracentrifugation and identified by electron microscopy and Western blotting. Then, un-exo and t-exo were used to treat a bovine macrophage cell line (BOSMAC), and the proliferative ability of BOSMAC cells was detected by an 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay and flow cytometry. The expression and phosphorylation levels of p38 were analyzed by q-PCR and Western blotting. The results showed that both exosome types exhibited the basic characteristics of exosomes. In BOSMAC cells treated with t-exo, significant inhibition of cell proliferation was observed, and the cell cycle progression was inhibited, while no difference was found between the un-exo and control groups. Only treatment with t-exo increased the expression and phosphorylation of p38, and the addition of the p38 inhibitor SB203580 abrogated the inhibition of BOSMAC cell proliferation by t-exo. Our results demonstrated that t-exo inhibited the proliferation of bovine macrophages by stimulating p38 MAPK and might interfere with immunity in dairy cattle. This finding may provide a new strategy for improving immunity and preventing breast-related diseases in dairy cows.