Extended Data Fig. 3: Disruption of the association of 2A1 B30.2 to 3A1 20.2 impairs γδ T cell responses to DMAPP and IPP.
From: Phosphoantigens glue butyrophilin 3A1 and 2A1 to activate Vγ9Vδ2 T cells
a, Cytotoxicity of Vγ9Vδ2 T cells towards BTN2A1WT and BTN2A1−/− MIA PaCa-2 cells (n = 6), which were pretreated with zoledronate (Zol) for 24 h. Data analysis: Two-way ANOVA with Šídák’s multiple comparisons test. P relative to the control. Error bars: SEM. b, ITC results for DMAPP and IPP binding to 3A1 B30.2. c, Cytotoxicity of Vγ9Vδ2 T cells towards BTN2A−/− MIA PaCa-2 (2A1/2A2 KO) cells stably expressing 2A1 WT or ΔC mutant variants (n = 3), treated with different concentration HMBPP/DMAPP/IPP. Data analysis: Two-way ANOVA with Šídák’s multiple comparisons test. P indicated the comparison between two groups at equal concentrations. Error bars: SEM. Representative of three independent experiments in HMBPP/DMAPP group. Representative of two independent experiments in IPP group. d, Analysis of the interactions between 2A1 B30.2 A chain and B chain reveal residues affecting γδ T cell responses. e, Flow cytometry analysis of His-tagged 2A1 mutant variants in BTN2A−/− 293T cells (n = 4, representative of two independent experiments), as detected by anti-His mAb. Error bars: SEM. f, SEC-MALS analysis of 2A1 B30.2R449A/R469A. The 2A1 B30.2R449A/R469A exists mainly as a monomer in solution. g, ITC results indicate that 2A1 B30.2R449A/R469A does not bind to 3A1 B30.2 in the presence of HMBPP.
