In macrophages, mitogen-activated protein kinases (MAPK) are critical regulators of both, mycobacterial replication and mycobacteria-induced cytokine formation. To segregate direct effects of MAPK function on mycobacterial replication from indirect, cytokine-mediated effects, we studied the growth of Mycobacterium avium strains in wild-type and tumor necrosis factor (TNF)-α- or interleukin (IL)-10-deficient bone marrow-derived murine macrophages. Using specific inhibitors of the p38- and the ERK1/2-MAPK pathways, we found that the use of SB203580 always reduced, whereas the presence of PD98059 always promoted, bacterial replication of highly virulent and intermediately virulent M. avium strains, independent of endogenous TNF-α or IL-10. The exogenous addition of TNF-α to TNF-α-deficient and wild-type M. avium-infected macrophages overrode the replication-reducing effect of SB203580, but not the replication-promoting effect of PD98059. In summary, our data demonstrate that a proper balance of MAPK activity is essential for macrophage control of M. avium growth, and that the ratio of the cytokines TNF-α and IL-10 can additionally modulate replication. Our findings indicate a novel therapeutic avenue for treating mycobacterial infections in particular by stimulating ERK1/2 or activating ERK1/2-dependent mechanisms in infected macrophages.