The aim of the present study was to determine the NO production by human cultured macrophages (m phi) and to compare the NO production between healthy subjects and patients with active pulmonary tuberculosis. The bioassay method was used for assessment of validation. Lipopolysaccharide (125 ng ml(-1))-activated m phi from healthy and diseased subjects released a substantial amount of NO. NO synthase inhibitor, N-G-nitro-L-arginine methyl ester, (0.1 mmol l(-1)) suppressed NO synthesis significantly in m phi of healthy subjects. Nitrite formation measured by the diazotization method in the supernatants taken from cultured m phi of tuberculous patients were significantly lower than the healthy subjects. The supernatants obtained in both subjects caused relaxations of guinea-pig aorta reversed by methylene blue (10 mu mol l(-1)). There was a significant difference between relaxations of healthy and diseased supernatants. Nitrite formation measured by the bioassay method in the supernatants taken from cultured m phi of tuberculous patients was significantly higher than the healthy subjects. It was concluded that NO production appeared to be decreased in tuberculosis. The reason for decreased production of NO in tuberculosis may be related to the interaction of several cytokines and/or eicosanoids by means of the disease related induction of immune reactions. (C) 1998 The Italian Pharmacological Society.