Nitric oxide (NO) and IL-12 are important mediators of the immune response to Leishmania major. In this study, the effects of L. major promastigotes, crude antigenic fraction (CAF) and its subfractions on NO production and IL-12 secretion by BALB/c mice peritoneal macrophages is investigated. The subfractions of CAF, namely, fractions 1, 2 and 3, that were in the molecular weight range of 97.4-66, 66-45 and below 45 kDa, respectively, were separated by SDS-PAGE. NO production was determined by using Griess reagent and IL-12 was measured by ELISA. It was found that NO production was stimulated by promastigotes but not by CAF or its subfractions. IL-12 secretion was stimulated by promastigotes, CAF and fraction 1 while fractions 2 and 3 did not have any effect.