CD4 T cells are important in the protective immune response against tuberculosis. Two mouse models deficient in CD4 T cells were used to examine the mechanism by which these cells participate in protection against Mycobacterium tuberculosis challenge. Transgenic mice deficient in either MHC class II or CD4 molecules demonstrated increased susceptibility to M. tuberculosis, compared with wild-type mice. MHC class II−/− mice were more susceptible than CD4−/− mice, as measured by survival following M. tuberculosis challenge, but the relative resistance of CD4−/− mice did not appear to be due to increased numbers of CD4− 8−(double-negative) T cells. Analysis of in vivo IFN-γ production in the lungs of infected mice revealed that both mutant mouse strains were only transiently impaired in their ability to produce IFN-γ following infection. At 2 wk postinfection, IFN-γ production, assessed by RT-PCR and intracellular cytokine staining, in the mutant mice was reduced by> 50% compared with that in wild-type mice. However, by 4 wk postinfection, both mutant and wild-type mice had similar levels of IFN-γ mRNA and protein production. In CD4 T cell-deficient mice, IFN-γ production was due to CD8 T cells. Thus, the importance of IFN-γ production by CD4 T cells appears to be early in infection, lending support to the hypothesis that early events in M. tuberculosis infection are crucial determinants of the course of infection.