Baseline expression of lymphatic and inflammatory genes in CD14+ blood monocytes from healthy donors

Blood was donated by four healthy volunteers with no known illness or malignancies. Mononuclear cells were separated from the blood by density gradient and CD14+ cells were isolated using anti-CD14 IgG-conjugated magnetic beads and magnetic separation (Miltenyi Biotec). We used an in-house array to analyze the gene expression of a total of 270 targets including 67 cytokines, 57 cytokine receptors, 18 chemokines, 11 chemokines receptors, 10 known TLR4 downstream targets, 64 other inflammation-related proteins, 12 genes expressed primarily in lymphatic endothelial cells, and 32 endothelial cell associated proteins. The results show a baseline expression in CD14+ PBMC in healthy individuals which can be used to detect changes in CD14+ PBMC population via transcript expression in individuals with illnesses and malignancies such as breast cancer. Transcript of CD14+ peripheral blood monocytic cells (PBMC) was analyzed by RT-qPCR gene expression profiling. CD14+ PBMC were isolated from four donors. For each specimen total RNA was isolated, cDNA synthesized, and 269 targets were analyzed by RT-qPCR.