Skip to main content
. 2022 Aug 26;17(9):2559–2571. doi: 10.1021/acschembio.2c00497

Figure 2.

Figure 2

Combination of cGAMP and CL075 drove IL-12-dependent Th1 polarization in human neonatal T cells. (A) Neonatal CBMCs were cultured in vitro for 96 h in the presence of polyclonal T cell activator α-CD3 (1 μg/mL) with or without CL075 (5 μM), cGAMP (25 μg/mL), or cGAMP + CL075, followed by IFNγ production evaluation by ELISA. (B) Example gating strategy for the quantification of CD4+ and CD8+ T cells. (C–H) CBMCs were cultured in vitro as in panel (A) but with the addition of a mitogen and cytokine blocker for the last 6 h. After stimulation, cells were harvested, stained (intracellular cytokine staining), and analyzed by flow cytometry to quantify the percentage of T cells producing IL-4, IL-17, and IFNγ. (I) CBMCs were cultured in vitro as in panel (A) but with the addition of human blocking Abs, αIFNAR2, or (J) αIL-12p40/70. After stimulation, the collected supernatant was evaluated via ELISA for IFNγ. Data were representative of two independent experiments. Statistical comparison employed test one or two-way ANOVA corrected for multiple comparisons; *p < 0.033, **p < 0.002 (n = 7 per group).