Mycobacterial crossreactivity of M. tuberculosis reactive T cell clones from naturally converted PPD positive healthy subjects

Mycobacterium tuberculosis reactive CD4⁺, CD8⁻ T cell clones were established from six naturally converted PPD positive healthy subjects by using whole bacilli as the primary stimulation antigen in vitro. Antigen specificity of the T cell clones was mapped by testing their proliferative response against a panel of pathogenic and environmental mycobacterial species. The crossreactivity patterns obtained showed that the T cell clones distributed along a spectrum from reactivity restricted to the M. tuberculosis complex to broadly crossreactive clones recognizing all mycobacterial species tested. Two of the T cell clones were able to discriminate between M. tuberculosis and M. bovis BCG, and importantly one of these clones was exclusively specific to M. tuberculosis. All of the CD4⁺ T cell clones tested, displayed MHC class II restricted cytotoxicity against macrophages pulsed with M. tuberculosis. In addition, some of these clones secreted GM-CSF upon antigen stimulation. The T cell clones described here represent relevant tools to identify and characterize target antigens of the immune response against M. tuberculosis with relevance to diagnosis and subunit vaccine design.