The anti-mycobacterial activities of IFN-gamma and TNF-alpha-treated murine peritoneal macrophages were determined. Resident macrophages pretreated with IFN-gamma or TNF-alpha for 2 days were infected with test organisms and subsequently cultured for up to 7 days. First, the early-phase growth of Mycobacterium tuberculosis (days 0-3) was strongly suppressed in IFN-gamma-treated macrophages, and progressive bacterial elimination was subsequently observed. Although TNF-alpha treatment of macrophages did not affect the early phase growth of organisms, bacterial killing was observed in the later phase of cultivation. Second, although IFN-gamma-treated macrophages killed M. avium during the first 3 days of culture, regrowth of the intracellular organisms was subsequently observed. TNF-alpha treatment of macrophages did not influence the mode of intracellular growth of M. avium. Third, IFN-gamma but not TNF-alpha enhanced production of reactive nitrogen intermediates (RNI) by macrophages infected with M. tuberculosis or M. avium, whereas both cytokines increased macrophage release of reactive oxygen intermediates (ROI). The present findings therefore show that IFN-gamma and TNF-alpha potentiated the anti-mycobacterial activity of murine peritoneal macrophages in different fashions. They also suggest that RNI played more important roles than did ROI in the expression of macrophage anti-mycobacterial, particularly anti-M. avium, activity.