T-cells' gene expression after high-dose melphalan. T-cells' gene expression after high-dose melphalan

T-cells' transduction efficacy by lentiviral transduction is diminished after high-dose melphalan. To investigate transcriptional changes regarding genes inhibiting viral entry, reverse transcription, nuclear transport and viral genome integration, we performed microarray transcription analysis. Overall design: Peripheral blood mononuclear cells of patients, who were treated with high-dose melphalan and autologous stem-cell transplantation, were isolated from peripheral blood via density centrifugation and subsequently activated by CD3/28-nanomatrix (Human TransAct, Miltenyi Biotech) and Interleukin-2. After 3 days, T-cells were isolated by cell-sorting (MoFlow Astrios, Beckman Coulter), and whole-RNA was extracted by Qiagen RNEasy Kit.