The contribution of lymphotoxin (LT)alpha in the host immune response to virulent Mycobacterium tuberculosis and Mycobacterium bovis bacillus Calmette-Guerin infections was investigated. Despite their ability to induce Th1 cytokine, IFN-gamma, and IL-12 pulmonary response, "conventional" LTalpha(-/-) mice succumb rapidly to virulent M. tuberculosis aerosol infection, with uncontrolled bacilli growth, defective granuloma formation, necrosis, and reduced pulmonary inducible NO synthase expression, similar to TNF(-/-) mice. Contributions from developmental lymphoid abnormalities in LTalpha(-/-) mice were excluded because hematopoietic reconstitution with conventional LTalpha(-/-) bone marrow conferred enhanced susceptibility to wild-type mice, comparable to conventional LTalpha(-/-) control mice. However, conventional LTalpha(-/-) mice produced reduced levels of TNF after M. bovis bacillus Calmette-Guerin infection, and their lack of control of mycobacterial infection could be due to a defective contribution of either LTalpha or TNF, or both, to the host immune response. To address this point, the response of "neo-free" LTalpha(-/-) mice with unperturbed intrinsic TNF expression to M. tuberculosis infection was investigated in a direct comparative study with conventional LTalpha(-/-) mice. Strikingly, although conventional LTalpha(-/-) mice were highly sensitive, similar to TNF(-/-) mice, neo-free LTalpha(-/-) mice controlled acute M. tuberculosis infection essentially as wild-type mice. Pulmonary bacterial burden and inflammation was, however, slightly increased in neo-free LTalpha(-/-) mice 4-5 mo postinfection, but importantly, they did not succumb to infection. Our findings revise the notion that LTalpha might have a critical role in host defense to acute mycobacterial infection, independent of TNF, but suggest a contribution of LTalpha in the control of chronic M. tuberculosis infection.