TCRs from Mtb-specific GLIPH2 groups vary in their capacity to recognize infected macrophages. (A–E) Bar graphs of mean (±SD) %CD69⁺ cells by flow cytometry (gated on CD4⁺ TCRβ⁺ Live-Dead^Lo) for SKW-3 cells transduced with representative TCRs from each of the indicated GLIPH2 groups 18 h after coculture with macrophages infected at MOI 1 or 4 (± α-HLA-DR/DP/DQ blockade), or left uninfected ± treatment with lysate or anti-CD3/CD28 bead stimulation. (F and G) Representative flow cytometry plots of CD69 and TCRβ expression of CD4⁺ SKW-3 cells transduced with the (F) R%SGGEAKNI or (G) RKQG%E TCRs in response to Mtb-infected or lysate-treated macrophages in separate experiments. (H and I) Bar graphs of mean (±SD) CD69 expression (of CD4⁺ TCRβ⁺) for the (H) RKQG%E and (I) SLRT%ET TCRs in response to infected macrophages. Data are representative of two to three independent experiments containing two to five replicates per condition. Statistical significance was determined using a Welch one-way ANOVA and Dunnett’s T3 post hoc test corrected for multiple comparisons; *P < 0.05, **P < 0.01, ***P < 0.001, n.s., not significant.