We focused on the major peripheral blood lymphocyte populations that may be involved in anti-tumor responses and negatively impacted by cancer, specifically CD8 T cells, CD4 T cells, B cells and CD56dim natural killer cells. The pure cell subsets were stringently sorted by flow cytometry from PBMC samples. Gene expression profiles of these cell populations from melanoma patients were compared to healthy controls. Keywords: gene expression profile
Overall design
The raw data set contained 48 arrays: 6 healthy and 6 melanoma arrays for each of the 4 cell types. Two of the arrays had quality issues due to background noise and were excluded, leaving us with 46 arrays.
On each array we used Cy3 to label a pool of two RNA samples from a pair of age and gender matched stage IV (American Joint Committee on Cancer) melanoma patients or from a pair of age and gender matched healthy donors. We used Cy5 to label the Total Lymphocyte Reference (TLR) RNA. The TLR is a common reference specifically created for this study using the total peripheral lymphocyte fraction from 20 healthy donors.
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