High fructose consumption aggravates inflammation by promoting Th1 and Th17 cell generations through the glutamine metabolism-dependent mTORC1 activation (PRJCA029873)

To investigate how fructose regulates Th1 and Th17 cell differentiation, we used bulk RNA sequencing (RNA-seq). Complete DMEM (cDMEM) was prepared for all cell cultures by adding 25 mM glucose or fructose to the glucose-free DMEM. CD4+CD25-CD62L+ naive T cells isolated from C57BL/6 mice were cultured in 48-well plates (2 x 10^5 per well) in the presence of plate-bound anti-mouse CD3 (1.5 µg/mL) and soluble anti-mouse CD28 (1.5 µg/mL), mouse IL-6 (50 ng/mL), TGF-ß1 (2 ng/mL), and mouse IL-12 (10 ng/mL). Cells were cultured at 37?C , 5% CO2, and collected for sequencing analysis three days later.