Hexokinase 2 is dispensable for T cell-dependent immunity

Hexokinase 2 (HK2) is one the most highly upregulated enzymes in glycolysis in activated T cells and cancer cells. We genetically abolished HK2 expression in T cells in vivo and infected mice with LCMV to induce T cell proliferation. CD8+ T cells from were sorted from spleens of WT and HK2 null mice 8 days after infection. Additionally, WT CD8+ T cells were compared to WT primary T cell acute lymphoblastic leukemia (T-ALL). T-ALL cells were sorted from spleens of mice 6 weeks after adoptive transfer of bone marrow that overexpresses mutant Notch1 (Notch1-deltaE). Whole RNA was extracted from 100k primary cells from sorted T cells and T-ALL using the RNAeasy kit (Qiagen) according to the manufacturer’s protocol. SMART-Seq v4 Ultra Low Input RNA Kit was used to generate full-length cDNA and Nextera XT DNA sample preparation kit was used to prepare final library. Libraries were sequenced on the NextSeq 500 with single-end sequencing (1x75bp). Differential gene expression analysis was performed between WT and and HK2 null (KO) T cells. Additionally, WT T cells were compared to WT T-ALL. Analysis was performed using edgeR package.