Pluto Bioinformatics

GSE150120: Transcriptom profiling of in vitro induced exhauted CD8+ T cells

Bulk RNA sequencing

By comparing transcriptome profiling of in vitro repeated stimulated cells with without stimulation and single peptide stimulated cells, to evaluated if the transcriptome profiling of repeated stimulated cells meet the features of exhausted cells. CD8+ T cells purified from OT-I mice were stimulated daily for five days with 10ng/ml OVA(257-264) peptide in the presence of IL-15 and IL-7. As controls, cells were either left without stimulation or stimulated only once with OVA(257-264) peptide for 48 hours and then washed and cultured without peptide for an additional 3 days. On day 5, cells were harvested. Live CD8+ T cells were sorted and RNAseq were performed after RNA isolation. It was found that the transcriptional prolife of the repeat peptide stimulated cells is clearly distinct to those of the single and non-stimulated cells. Among the upregulated genes, multiple inhibitory receptors encoding genes were presented on the top of this list. The genes for the markers of the terminal differentiated effector cells, like GzmB, GzmC and Prf1 were also significantly upregulated on the repeat peptide stimulated cells. The expression pattern of exhaustion specific transcription factors were identified in repeat stimulated cells. In vitro repeat peptide stimulation results in the transcriptional changes of in vivo exhausted CD8+ T cells from chronic viral infections which serve as the benchmark of CTL exhaustion. SOURCE: Peter,D,Katsikis (p.katsikis@erasmusmc.nl) - IIG Erasmus University Medical Center