Polyunsaturated fatty acids (PUFAs), known modulators of the immune response, are the source of essential fatty acids in total parenteral nutrition-dependent patients. Critically ill infants on TPN have an increased incidence of sepsis, and lipid emulsions depress various immune functions. Recent studies have demonstrated that PUFAs induce apoptosis in various tissue cells in vitro and ex vivo. The susceptibility of neonatal monocytes, as major early effector cells in the host response to sepsis, to PUFA-mediated apoptosis and the mechanisms associated with PUFA-induced apoptosis were investigated. Both n-3 and n-6 PUFAs induced rapid, dose-dependent cell death in purified monocytes. Polyunsaturated fatty acids induced significant activation of upstream caspases 8 and 9 as well as caspase 3. The PUFA treatment resulted in a 4-fold increase in oxidative stress and a loss of monocyte mitochondrial potential compared with carrier controls (P < .05). The addition of cyclosporin, which blocks the development of mitochondrial transition pores, completely abolished the proapoptotic effects of PUFAs. Although Trolox (Sigma Aldrich) reduced PUFA-induced intracellular oxidative stress in neonatal monocytes, apoptosis was not blocked by this potent antioxidant. The data identify PUFAs as potent inducers of monocyte apoptosis, which can occur independently of the induction of oxidative stress, by using a mitochondrial dependent pathway. The TPN-dependent infant may be particularly sensitive to such PUFA effects, having a relatively poor capacity to both use and clear PUFAs.