Imiquimod

TALL-104 human leukemic CTLs were maintained as described previously ^{9 (link)}. Early experiments were conducted in complete culture medium. Later experiments were in culture medium without IL-2. Complete culture medium contains 40mL heat-activated fetal bovine serum, 4mL Pen-Strep, 8mL L-Glutamine, 80 μL IL-2, and 348mL Iscove’s modified Dulbecco’s medium. Dynabeads polystyrene beads coated with anti-CD3 or anti-CD8 monoclonal antibody (Thermo Fisher, Waltham, MA) were washed in medium according to the manufacturer’s protocol, and were added to the cell suspension at a ratio of ~one bead per cell. Alexa647-conjugated anti-LAMP1 antibody (0.5 μg/mL final concentration) was purchased from BD Pharmingen (San Jose, CA). Phorbol 12-myristate 13-acetate (PMA) was from Alexis Biochemicals (San Diego, CA). Thapsigargin (TG) was from AdipoGen (San Diego, CA). Cells were incubated with imiquimod (Adipogen) San Diego, California) at 10 uM or PMA at 100 nM for indicated time. The National Cancer Institute’s Diversity Set V (https://wiki.nci.nih.gov/display/NCIDTPdata/Compound+Sets) was used for screening, and powder resupply was from NCI. Compounds were prepared in DMSO at a final concentration of 10μM.