Panel A shows a volcano plot with x-axis: Log2 fold change and y-axis: negative Log10 P-value. Differentially expressed genes are compared between P1 and healthy controls. Panel B shows a heatmap with x-axis: samples and y-axis: Th17-related genes. Expression patterns of Th17-associated genes are compared between P1 and healthy controls. Panel C shows scatter plots with x-axis: P1 and healthy controls and y-axis: Log2 count per million reads. Expression levels of RORC, MAF, SOX13, LEF1, IL23R, IL9R, ADAM12, IL17RE, and CCR6 are compared between groups. Panel D shows a heatmap with x-axis: samples and y-axis: Th9-related genes. Expression profiles of Th9-associated genes are compared between P1 and healthy controls. Panel E shows histograms with x-axis: interleukin-9 expression and y-axis: normalized to mode. Interleukin-9 production is assessed under unstimulated, HMBPP-stimulated, and PMA/Ionomycin-stimulated conditions. Panel F shows a scatter plot with x-axis: Vehicle, HMBPP, and PMA/Ionomycin conditions and y-axis: interleukin-9 mean fluorescence intensity. Interleukin-9 production is compared between P1 and healthy controls.
P1-derived TCR γδ cells express fewer Th17-related genes than HCs and have a Th1 profile. (A) Volcano plot of genes differentially expressed between TCR γδ cells derived from P1 or HCs. Genes represented as red dots have a P <0.05, and genes represented as blue dots have a P >0.05. Marked genes are linked to TH17 differentiation (RORC, MAF, SOX13, IL23R, IL12RB1, IL17RE, KIT, CCR6, LEF1, TIAM1, ITGA2) or to STAT pathways (STAT1, STAT6, SOCS1, SOCS2, IL-9R). (B) Heatmap of genes involved in Th17 differentiation and STAT5B regulation (RORC, MAF, CCR6) differentially expressed between P1 and HCs. (C) mRNA expression level of RORC, MAF, SOX13, LEF1, IL-23R, IL-9R, ADAM12, IL-17RE, and CCR6 from overnight-rested P1 (red circles)– and HC (black circles)–derived TCR γδ cells. 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 without a normal distribution, the Mann–Whitney test was used. **P < 0.01, ***P < 0.001, ****P < 0.0001. (D) Heatmap of genes related to the Th9 profile differentially expressed between P1 and HCs. (E) 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-9 produced by TCR Vδ2 cells from HC105 (dashed black line), HC106 (black line), and P1 (red line). The data correspond to one experiment. (F) Intracellular cytokine production of IL-9 by TCR Vδ2 cells from P1 (red circles) and HCs (black circles). MFI values are represented for IL-9. CPM, count per million reads.