Figure 5.

ThGM cells express a unique set of transcription factors. Human ThGM, Th1, and GM-CSF⁺Th1 (Th1-GM⁺) cells were isolated from total T_M cells of PB of healthy donors (n = 6 subjects) using a combination of FACS sorting and cytokine capture assay. RNA was extracted and analyzed using RNA-seq analysis. (A) Bar-graph showing enriched pathways for ThGM-specific genes. (B) Heatmap showing expression levels of ThGM- and Th1-specific TFs, chemokines, and surface molecules. (C) RNA expression of TWIST1, TRERF1, MSC, and PPARG by FACS-sorted naïve CD4⁺, Th1, and ThGM cells were determined by RT-PCR (n = 4 subjects). (D) Representative flow cytometry histograms for TWIST1, TRERF1, MSC, and PPARG expression by FACS-sorted naïve CD4⁺, Th1, and ThGM cells. (E) Human naïve CD4⁺ T cells were differentiated into ThGM cells and RNA was extracted at several time points. mRNA levels for TWIST1, TRERF1, MSC, and PPARG were quantified by RT-PCR. Data shown are mean ± SEM. P-values were calculated using unpaired Student’s t-test with Bonferroni’s correction; *p < 0.05, **p < 0.01.