Panel A shows scatter plots of various transcripts in gamma delta T cells of healthy controls versus P1. The y-axis represents Log2 CPM (count per million reads), and significant differences are indicated by asterisks. Panel B presents similar scatter plots for ex vivo TCR gamma delta cells. Panel C displays histograms of intracellular staining for various cytokines (IL-4, IL-10, IL-13, IL-17A, IL-22, IL-2, IFN-, TNF-, and GM-CSF) produced by TCR V delta 2 cells from healthy controls and P1 under different stimulation conditions. Panel D summarizes the intracellular cytokine production, showing MFI values.
Comparison of gene expression in TCR γδ cells and cytokine profiling in P1 vs. HCs. (A) mRNA expression level of RORC, MAF, SOX13, LEF1, IL-23R, IL-9R, ADAM12, IL-17RE, and CCR6 from P1 (red circles)– and HC (black circles)–derived TCR γδ cells stimulated with IL-2 (100 U/ml) for 4 h. Expression levels are shown as Log2 CPM. Normality was assessed using the Shapiro–Wilk test. For normally distributed data, an unpaired t test was used. For data that were not normally distributed, the Mann–Whitney test was used. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. (B) mRNA expression level of RORC, MAF, SOX13, LEF1, IL-23R, IL-9R, ADAM12, IL-17RE, and CCR6 from ex vivo P1 (red circles)– and HC (black circles)–derived TCR γδ cells. Expression levels are shown as Log2 CPM. (C) Activation of TCR Vδ2 cell lines challenged for 12 h with THP-1 cells exposed to a vehicle (left plots) or 4 nM of HMBPP (middle plots). T cells were also exposed to PMA/ionomycin to induce strong cell activation (right plots). Histograms of intracellular staining of IL-4, IL-10, IL-13, IL-17A, IL-22, IL-2, IFN-γ, TNF-α, and GM-CSF cytokines produced by TCR Vδ2 cells from HC105 (dashed black line), HC106 (black line), and P1 (red line). (D) Summary of intracellular cytokine production of IL-4, IL-10, IL-17A, IL-22, and GM-CSF by TCR Vδ2 cells from P1 (red circles) and HCs (black circles). MFI values are represented for IL-4, IL-10, IL-17A, IL-22, and IL-2. Percentages are represented for IL-13+, IFN-γ+, TNF-α+, and GM-CSF+ cells. The data correspond to one experiment. CPM, count per million reads.