Expression data ex vivo CD4 T cells systemic lupus erythematosus patients

Compare ex vivo unstimulated CD4 T cells from SLE and HC female patients in an ethnically mixed cohort for transcriptional differences. Collection of blood and negative isolation for CD4+ T cells from 8 patients of varying disease activity, all females, and matched healthy controls. 3 tubes of 10mL each were taken and isolated for CD4 T cells by Rosette Sep antibodies and density centrifugation. These antibodies deplete CD8, CD16, CD19, CD36, CD56, CD66b, gamma/delta TCRs, by crosslinking to glycophorin A on red blood cells. Isolated cells were immediately lysed in Qiagen's buffer RLT with 0.1% 2-mercaptoethanol and stored at -80C until further isolation with Qiagen RNeasy mini kit. RNA was quantified with a nanodrop machine and 200 ng for each sample was used for fragmenting and labeling targets according to affymetrix recommendations for the HTA 2.0 microarrays. Scanning was performed with GeneChip Scanner 3000 7G, and washed and stained with the GeneChip Fluidics Station 450.