We have investigated the relationship between cortisol and dehydroepiandrosterone (DHEA) levels and the immune response to mycobacterial antigens in peripheral venous blood, from a male population of active tuberculosis patients and age‐matched healthy controls of the same sex (HCo). Peripheral blood mononuclear cells were cultured for 36 or 96 h with whole sonicated Mycobacterium tuberculosis (WSA) for measurement of proliferation, interferon gamma (IFN‐γ) and interleukin‐10 (IL‐10) in culture supernatants. Comparisons on the in vitro mycobacterial‐driven immune responses demonstrated that TB patients had a higher IL‐10 production, a decreased lymphoproliferation and a trend to reduced IFN‐γ synthesis, in relation to HCo. Active disease was also characterized by increases in the plasma levels of glucocorticoids (GC) and reduced concentrations of DHEA which resulted in a higher cortisol/DHEA ratio respect the HCo group. Plasma DHEA levels were positively correlated with IFN‐γ values. An inverse correlation was found between the cortisol/DHEA ratio and IFN‐γ levels. Novel evidence is provided showing that the balance between cortisol and DHEA is partly responsible for the immune perturbations seen in TB patients.