Transcriptome analysis of blood V?9 Vd2 gdT cells upon TCR and Notch signalling

We utilized RNA-seq to identify differential gene expression in V?9 Vd2 gdT cells isolated from human PBMCs upon TCR stimulation. We find that ~30% genes are commonly up- and down-regulated between different phosphoantigen treatments (p<0.05). Inhibition of Notch signaling alongside TCR activation results in downregulation of many effector genes in V?9 Vd2 T cells. Overall design: PBMCs from healthy donors were subjected to MACS based gdT cell isolation. The isolated cells (~1 million each) were either untreated, treated with rIL2 only, or a combination of rIL2+IPP, rIL2+HDMAPP and rIL2+anti-CD3 for T cell activation. Notch signaling inhibitor treatment was done using GSI-X along with TCR activation. Three replicates were sequenced for all the 8 treatment samples.