Polymicrobial infections are common in wound infections, especially in diabetic lower-limb wounds. Wounds infected with multiple pathogens are often associated with worse prognoses, delayed recovery, and higher complexity treatments. Despite their clinical significance, little is known about the mechanistic interactions between the polymicrobial community and host immune cells. Using Staphylococcus aureus and Enterococcus faecalis, two pathogens commonly isolated from diabetic wounds, we established in vitro and in vivo models to investigate the responses of neutrophils upon polymicrobial infection and their impact on bacterial clearance. Upon exposure to E. faecalis in vitro, neutrophils increased intracellular ROS production but did not induce neutrophil extracellular trap (NET) formation, and chemical inhibition of ROS leads to significantly impaired E. faecalis killing. Co-infection of S. aureus with E. faecalis resulted in a significant reduction both intracellular ROS levels and E. faecalis clearance, suggesting that S. aureus can inhibit intracellular ROS production which promotes E. faecalis survival. Conversely, upon exposure to S. aureus, neutrophils underwent NETosis in the absence of detectable intracellular ROS. Co-infection with E. faecalis resulted in a significant reduction of S. aureus-induced NET formation, indicating that E. faecalis can interfere with S. aureus-induced NETosis. Using an in vivo mouse wound model, we also observed that co-infection is associated with augmented bacterial burdens for both E. faecalis and S. aureus, compared to either single species infection. Ongoing work will determine whether the in vivo synergy in bacterial burden that we observe upon co-infection arises via cross-species neutrophil modulation. Together these results suggest that optimal pathogen-specific neutrophil responses can be subverted in a reciprocal manner during mixed species infection, thus resulting in attenuated neutrophil-mediated clearance and augmented bacterial growth of both co-infecting organisms.