NADPH oxidase-derived reactive oxygen species production activates the ERK1/2 pathway in neutrophil extracellular traps formation by Streptococcus agalactiae isolated from clinical mastitis bovine

Abstract

Streptococcus agalactiae (S. agalactiae) continues to be challenging for milk quality in some countries and leads to huge economic losses. A large number of neutrophils are recruited into inflammatory foci when S. agalactiae infection occurs, and most studies have focused on the interaction between neutrophil extracellular traps (NETs) and this bacterium in the context of human pathogenicity. However, there is little information on the NET formation mechanism induced by S. agalactiae in the context of bovine mastitis. Here, neutrophils isolated from BALB/c mice were infected with S. agalactiae SAG-FX17, and NET formation was evaluated. SAG-FX17 could induce NADPH oxidase-derived reactive oxygen species (NOX-ROS)-dependent NET formation, and 21.8% of bacteria could be eliminated by NETs via NET DNA and associated proteins. SAG-FX17 could induce the phosphorylation of p38 MAPK, ERK1/2 MAPK, and JNK/SAPK in neutrophils. However, only ERK1/2 MAPK was shown to play an important role in SAG-FX17-induced NET formation. Importantly, NOX-ROS production occurs upstream of ERK1/2 MAPK activation and then induces NET release. ERK1/2 MAPK phosphorylation can, in turn, enhance NOX-ROS generation, which further contributes to NET release and bacterial elimination. This study provides evidence of the molecular mechanism underlying serotype Ia S. agalactiae SAG-FX17-induced NET formation and the interaction between bacteria and NETs, and these findings will increase our knowledge about bacterial mastitis in dairy cattle and contribute to the prevention and clinical treatment of bovine mastitis.