Expression data from C57/Bl6 Wt mice CD4+ T cells after activation with anti-CD3 plus Wt APC or OX40L-Tg APC

OX40 is a T cell costimulatory molecule in the tumor necrosis factor receptor (TNFR) superfamily. Unlike other costimulatory molecules, OX40 is not expressed by naive T cells, but its expression is rapidly induced following T cell activation, and OX40 costimulation plays a critical role in cell survival, proliferation, and generation of memory cells. We used microarrays to detail the global programme of gene expression of activated CD4 T cells underlying OX40 costimulation and identified distinct classes of up-regulated genes after OX40 costimulation. Naive CD4+ T cells (CD62LhighCD44lowFoxp3−) were sorted from C57/Bl6 Foxp3-eGFP reporter mice, and activated in vitro with soluble anti-CD3 mAb and mitomycin C-treated Wt APCs or OX40L-Tg APCs in 96-well round bottom tissue-culture plates. Forty-eight hours later, living CD4+ T cells were sorted from cultures by flow cytometry. Total RNA was extracted with RNeasy mini kit (Qiagen). Complementary DNAs (cDNAs) were then synthesized using Superscript III kits (Invitrogen), hybridization of the samples with Affymetrix mouse arrays was then performed at the Harvard Medical School Facility according to the manufacturer's protocol.