Expression data from comparison of wild type and cereblon knock out murine T-cells

Thalidomide and other immunomodulatory drugs are effective therapies for certain hematological cancers where they kill tumor cells and stimulate T-cell immunity. These compounds mediate their functions by binding to and altering substrate specificity of cereblon (CRBN), an E3-ubiquitin ligase component of the DDB1/Cul4/Rbx1 complex3-5. Here, we report the results of crbn genetic deficiency in a mouse, which revealed CRBN functions as a suppressor of T-cell receptor (TCR) activation. We used microarrays to analyze the cereblon regulated gene expression changes following activation of wild type or cereblon knock out murine T-cells. Splenic T-cells were isolated from C57BL6 wild type or cereblon knock out mice using negative magnetic bead selection (Pan T cell Kit II, Miltenyi) and grown in complete RPMI-10 media. Purified T-cells were then collected in their naïve, unactivated state or treated with anti-CD3 (5µg/ml plate bound) and anti-CD28 (1µg/ml soluble) for 12 hours. In total, 11 biological samples were divided into four groups based on two genotypes and two activation states: (1) wild type naïve n=3, (2) crbn-/- naïve n=3, (3) wild type activated n=2, (4) crbn-/- activated n=3.