Bovine aorta endothelial cells (BAECs) were used to determine the effect of ketone bodies and glucose on in vitro interferon (IFN), tumor necrosis factor (TNF) and nitric oxide (NO) production. BAECs were incubated for 4 and 24h with the ketone bodies: 3.8mmol/l beta-hydroxybutyrate (BHB), 1mmol/l acetoacetate (AcAc) and 5. 2mmol/l acetone (Ac), used separately or in a mixture together with cytokine inducers: Newcastle disease virus (NDV) and lipopolysaccharide (LPS). BHB alone (but not AcAc or Ac) and a mixture of ketone bodies caused a significant decrease in IFN titers induced by NDV and LPS and in TNF titers induced by LPS. Glucose used at concentrations of 5.55, 3.33 and 1.66mmol/l did not influence cytokine production.NO measured by the nitrite content in culture medium was released spontaneously from BAECs. A slight enhancement of NO release was observed after infection of BAECs with NDV; however, treatment with LPS caused inhibition of the release. The mixture of ketone bodies used with NDV or LPS enhanced NO release. However, when cells were incubated in the medium with 1. 66mmol/l glucose (mimicking low plasma glucose level in ketotic cows) a significant decrease in NO release was observed. This enhancing effect of ketone bodies and inhibition by low glucose in the final effect balanced each other, and the amounts of NO released in the medium with 1.66mmol/l glucose and with the mixture of ketone bodies resembled those produced at 3.33mmol/l glucose without ketone bodies. The significance of these effects of ketone bodies and glucose concentrations on cytokine and NO production in the immunity of ketotic cows has been discussed.