Lipopolysaccharide primes macrophages to increase nitric oxide production in response to Staphylococcus aureus.

Sepsis, the leading cause of death in intensive care units, is associated with overproduction of nitric oxide (NO). The mechanism concerning the NO production in the sepsis caused by both Gram-negative and Gram-positive bacteria is largely unknown. The present study examines the effect of lipopolysaccharide (LPS) on Staphylococcus aureus-induced NO production in macrophages. In the naïve murine macrophage cell line RAW264.7, heat-killed Staphylococcus aureus (HKSa) induced a significant NO production at a high concentration (100 microg/ml). However, pretreatment of the cells with increasing concentration of LPS (10-50 ng/ml) resulted in induction of NO production by HKSa even at the doses of 1 and 10 microg/ml. The expression of inducible NO synthase (iNOS) in response to HKSa was also enhanced by LPS pretreatment, suggesting that LPS priming NO production is due to the enhancement of iNOS expression. We examined whether protein kinase C (PKC), mitogen-activated protein kinases (MAPKs) and calcineurin signaling pathways are involved in the priming effects of LPS. It was found that the PKC inhibitor Gö6976, the p38 inhibitor SB203580 and the calcineurin inhibitor cyclosporine A significantly reversed the priming effects of LPS on HKSa-induced NO production and iNOS expression. In contrast, the ERK1/2 inhibitor PD98059 did not block the induction of priming by LPS. These data support the hypothesis that LPS primes macrophages for enhancement of HKSa-induced NO production, and indicate that PKC, p38 and calcineurin might be involved in the LPS-induced priming.