Down-regulation of NOD1 in neutrophils of periparturient dairy cows

Abstract

Increased susceptibility of periparturient dairy cows to infectious diseases has been linked to neutrophil dysfunction. Cytoplasmic nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs) mediate numerous aspects of innate immunity in humans and experimental animals. The authors hypothesized that neutrophils in periparturient cows have reduced NLR expression, resulting in declined immune responses to NLR activation. In this study, blood neutrophils from periparturient and mid-lactating cows were isolated and assessed for mRNA and protein expression of NOD1 and NOD2. Neutrophil phagocytosis and oxidative burst activity following NLR agonist-induced activation were also investigated. Periparturient cows demonstrated weak-to-absent NOD1 mRNA expression along with a significant reduction in NOD1 protein in their neutrophils. However, no difference in NOD2 protein expression was observed between groups although NOD2 mRNA abundance in periparturient cows was significantly reduced. Phagocytosis and oxidative burst activity measured by production of reactive oxygen species (ROS) were diminished in periparturient cows. NOD1 activation enhanced phagocytosis and ROS production in both isolated cell populations, regardless of the lactation stage. Nevertheless, NOD1-activated neutrophils showed reduced phagocytosis and ROS generation in periparturient cows with respect to mid-lactating subjects. Collectively, our findings suggest that NOD1 down-regulation in neutrophils may be a mechanism involved in periparturient neutrophil dysregulation.