A proteome-wide screen of 20,610 Mtb-derived peptides in 21 patients mid-treatment for active tuberculosis (ATB) identified 137 unique T cell epitopes recognized by IFNγ. Of these, 16% were recognized by multiple participants, predominantly derived from cell wall and cell processes antigens. The study found differential recognition of antigens between ATB participants and interferon-gamma release assay (IGRA +/−) individuals. An ATB-specific peptide pool (ATB116) was developed, consisting of epitopes exclusively recognized by ATB participants. This pool can distinguish patients with pulmonary ATB from IGRA +/− individuals from various geographical locations with a sensitivity of over 60% and a specificity exceeding 80%. The study identified infection stage-specific epitopes and antigens for potential use in diagnostics and measuring Mtb-specific immune responses. The results suggest that different patterns of antigenic ORFs might be associated with ATB vs. healthy IGRA+ participants. The study also found that the ATB-specific T cell epitope pool, ATB116, can distinguish ATB from healthy IGRA+ participants, with high sensitivity and specificity. The study also showed that the response to ATB116 changes during treatment for ATB, indicating that the response against ATB116 is differentially affected depending on the specific cytokine measured. The study highlights the diagnostic potential of ATB116 as a useful tool in identifying cases of active tuberculosis. The results suggest that the ATB-specific T cell epitope pool, ATB116, may be useful in identifying individuals at risk of developing active TB. The study also found that the response to ATB116 changes during treatment for ATB, indicating that the response against ATB116 is differentially affected depending on the specific cytokine measured. The study highlights the diagnostic potential of ATB116 as a useful tool in identifying cases of active tuberculosis.A proteome-wide screen of 20,610 Mtb-derived peptides in 21 patients mid-treatment for active tuberculosis (ATB) identified 137 unique T cell epitopes recognized by IFNγ. Of these, 16% were recognized by multiple participants, predominantly derived from cell wall and cell processes antigens. The study found differential recognition of antigens between ATB participants and interferon-gamma release assay (IGRA +/−) individuals. An ATB-specific peptide pool (ATB116) was developed, consisting of epitopes exclusively recognized by ATB participants. This pool can distinguish patients with pulmonary ATB from IGRA +/− individuals from various geographical locations with a sensitivity of over 60% and a specificity exceeding 80%. The study identified infection stage-specific epitopes and antigens for potential use in diagnostics and measuring Mtb-specific immune responses. The results suggest that different patterns of antigenic ORFs might be associated with ATB vs. healthy IGRA+ participants. The study also found that the ATB-specific T cell epitope pool, ATB116, can distinguish ATB from healthy IGRA+ participants, with high sensitivity and specificity. The study also showed that the response to ATB116 changes during treatment for ATB, indicating that the response against ATB116 is differentially affected depending on the specific cytokine measured. The study highlights the diagnostic potential of ATB116 as a useful tool in identifying cases of active tuberculosis. The results suggest that the ATB-specific T cell epitope pool, ATB116, may be useful in identifying individuals at risk of developing active TB. The study also found that the response to ATB116 changes during treatment for ATB, indicating that the response against ATB116 is differentially affected depending on the specific cytokine measured. The study highlights the diagnostic potential of ATB116 as a useful tool in identifying cases of active tuberculosis.