CTCL and T-ALL cell lines were treated with various concentrations of SGN-CD70A or PBS for 72 hours. The cell proliferation was measured by CellTiter-Glo (Promega, Leiden, Netherlands), and apoptosis was analyzed by the annexin-V/PI assay.
Primary tumor cells from PDX mice were cultured in complete media, RPMI1640 with 20% human AB serum (MP, Solon, OH). For the proliferation assays, primary tumor cells from PDXs were stimulated to grow with CD2/CD3/CD28 activation beads and incubated with hIL-2 (500 U/mL) and hIL-7 (1000 U/mL) (Miltenyi Biotec) at an optimal cell density of 5 to 10 × 105 cells/mL. After 24 or 48 hours of activation, the cells were treated with various concentrations of SGN-CD70A as indicated. The proliferation of primary tumor cells was determined by Real-Time Glo (Promega) at 24, 48, and 72 hours after drug treatment. Apoptosis assays using primary tumor cells were performed as described previously.20 (link) Briefly, primary tumor cells were harvested from the spleens of PDX mice and cultured in a complete medium with 250 U/mL of hIL-2 without activation beads. At 24 hours, cells were treated with various concentrations of SGN-CD70A or ADC-IgG control. After 72 hours of treatment, cells were analyzed by the annexin-V/PI assay.
Primary tumor cells from PDX mice were cultured in complete media, RPMI1640 with 20% human AB serum (MP, Solon, OH). For the proliferation assays, primary tumor cells from PDXs were stimulated to grow with CD2/CD3/CD28 activation beads and incubated with hIL-2 (500 U/mL) and hIL-7 (1000 U/mL) (Miltenyi Biotec) at an optimal cell density of 5 to 10 × 105 cells/mL. After 24 or 48 hours of activation, the cells were treated with various concentrations of SGN-CD70A as indicated. The proliferation of primary tumor cells was determined by Real-Time Glo (Promega) at 24, 48, and 72 hours after drug treatment. Apoptosis assays using primary tumor cells were performed as described previously.20 (link) Briefly, primary tumor cells were harvested from the spleens of PDX mice and cultured in a complete medium with 250 U/mL of hIL-2 without activation beads. At 24 hours, cells were treated with various concentrations of SGN-CD70A or ADC-IgG control. After 72 hours of treatment, cells were analyzed by the annexin-V/PI assay.
