Expression data from pmel1 CD8+ T cells stimulated with agonist anti-CD27, blockade of PD-1/L1 or both together

CD8+ T cells are regulated by inhibitory and activatory receptors. We have investigated the influence of enforced CD27 stimulation compared with blockade of PD-1 and the combination of both agents together. Anti-CD27 and PD-1 blockade combined to enhance CD8+ T-cell accumulation, effector protein expression and tumor therapy. This array was used to further characterize the transcriptional changes in CD8+ T cells that drive this differentiation. Two million Thy1.1+CD8+ T cells from pmel1 mice were adoptively transferred to naive congenic recipient mice. Recipients (in groups of 3) were then treated with 100ug hgp100 peptide i.v. delivered with a total of 400ug antibodies comprising 200ug agonist anti-CD27 (AT124), with 100ug anti-PD-1 (RMP1-14) and 100ug anti-PD-L1 (10F.9G2) (termed 'combination'); alternatively mice were treated with peptide+ AT124 (anti-CD27), or peptide + anti-PD-1/anti-PD-L1 (anti-PD-1/L1) mixed with 200ug appropriate isotype control antibodies. On day 4 relative to peptide injection CD8+ cells from spleens were isolated from individual mice using negative selection MACs kits, and then Thy1.1+CD8+ cells enriched to >99% purity by FACs. RNA was extracted from a minimum of 0.4x10^6 cells using RNeasy kits and gDNA eliminator columns (Qiagen). Samples were stored at -80C and subsequently processed for array by AROS (Applied Biotechnology (Denmark).