Granted that NETosis and NE release was significantly elevated in GDM, our next task was to gain insight into the factors promoting this change in neutrophil activity. A number of reports have indicated that circulatory levels are elevated in cases with GDM and that the potent pro-inflammatory cytokine TNF-α may assist in promoting a pro-diabetic state (45, 46). Our results indicated that circulatory TNF-α levels were indeed elevated in cases with GDM when compared to matching controls (Figure 2C). Since TNF-α is a potent factor for neutrophil activation and NETosis (47), we examined the effect of plasma from GDM cases and normal healthy control pregnancies. This analysis indicated that plasma from GDM cases promoted a greater degree of NETosis on isolated control neutrophils than comparable plasma from healthy pregnant women (Figure 2B). To confirm that TNF-α played a key role in mediating this pro-NETotic effect, we used infliximab, a specific TNF-α antagonist. Such treatment significantly diminished the pro-NETotic effect of GDM plasma on isolated normal neutrophils (Figure 2B).