Regulation of T cell gene expression and function by the steroid hormone precursor Pregnenolone

We report the transcriptional changes in primary T cells expanded from peripheral blood mononuclear cells (PBMCs) upon treatemnt with the steroid precursor, pregnenolone. Overall design: Expanded T cells were non-activated or activated through the T cell receptor with CD3/CD28 Dyna Beads for 24 hours. They were subsequently treated or untreated with 200 µM of pregnenolone for 2 or 24 hours. We additionally included a "basal" condition that involved no activation or treatment at 0 hours (time of activation). All nineconditions included 2-4 biological replicates, whereby the same expanded T cells were activated/non-actived and treated/untreated in separate wells per condition, yielding a total of 33 samples. RNA was extracted from each sample, and library preparation was performed, whereby some samples had library preparation done in duplicates (technical rep). cDNA libraries were then seuqenced.