GLIPH2 refines estimates of Mtb-specific CD4 ⁺ T cell recognition of infected macrophages. (A) Venn diagram indicating the number of GLIPH2 groups containing TCRs linked to responses to infected macrophages (green, stars), MTB300 (added to infected macrophages or to PBMCs) (blue, squares), control peptide megapools added to PBMCs (red), or lysate (added to infected macrophages) (yellow, triangle). (B) Pie charts of percentage (and number) of remaining GLIPH2 groups that respond to infected macrophages (green; sum of star groups from A) or MTB300 only (blue; sum of square groups from A). (C) Pie charts of GLIPH2 groups (left) and corresponding unique TCRβs (right) after removing GLIPH2 groups containing TCRs linked to viral antigen responses. (D) Bar graph of GLIPH2 groups (x axis) estimated to be Mtb-specific, rank-ordered by the sum of the highest number of TCR copies per experimental condition (y axis). Responses to infected macrophages (green), MTB300 only (blue), or both (blue stripes) are indicated. “%” indicates any amino acid substitution. (E) Pie charts comparing responses to infected macrophages (green) or MTB300 peptides only (blue) for GLIPH2 groups (left) containing unique TCRβs (right) contributed by ≥2 participants. Data were generated from a combined list of expanded TCRs from all experimental conditions (10 experiments; 10 LTBI and 6 non-LTBI participants).