Cutaneous T-cell Lymphoma cells included 2 MF (HH and MYLA) and 2 SS (HUT78 and SeAx) cell lines that were tested as mycoplasma free. Patient-derived SS samples (identified as SZ #1–4) were obtained from fresh peripheral blood mononuclear cells of selected SZ patients with high tumor burden (representing 90–95% of CD4+ SZ cell population according to current morphologic and/or phenotypic diagnostic criteria). Mononuclear cells were recovered by Ficoll (GE Healthcare, Princeton, NJ) gradient separation. CD4+ T cells were purified by positive selection using immune-magnetic beads (Miltenyi Biotec, Bergisch Gladbach, Germany), cultured at a density of 1 × 106 cells/ml and then activated and expanded in vitro using anti-CD3/CD28 beads (Life Technologies, Norway).
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