[show abstract][hide abstract] ABSTRACT: This study was conducted to evaluate the anti Mycobacterium tuberculosis effects of Ocimum sanctum directly and in macrophage culture which were infected with Mycobacterium tuberculosis before treatment with different concentrations of this plant. Suspensions of bacteria were prepared in 7H9 broth and after Macrophage culture, cell suspensions of the M.tuberculosis were added to the attached macrophages. Adherent monolayers was disrupted and bacterial suspensions were serially diluted and plated onto Middle brook 7H10 agar plates. Colonies were counted under a dissecting microscope and reported as CFU. For each culture dilution, six replicate samples were plated and the mean number of colonies was calculated and then Intracellular and extra cellular killing of Ocimum sanctum extracts were measured by colony counting. Present findings showed that in a defined laboratory and macrophage culture, Ocimum sanctum has a potent anti- Mycobacterium tuberculosis effects both directly and in infected macrophage culture. In this study we confirmed anti-tuberculosis effects of different concentration of O. sanctum extracts in vitro and in Macrophage culture but key components of anti tuberculosis action of these extracts and their mechanisms of actions must be discovered in future researches.