Memory T cells from poopled male and female CD57BL/6 mouse
Total CD8 T cells were negatively enriched using Dynabeads Untouched mouse CD8 cells kit (Invitrogen) from inguinal, brachial, axillary and cervical lymph nodes. For in vitro differentiation, naive or total CD8 T cells or OT-I CD8 T cells (1 x 1e+6 cells/ml) were activated in vitro for 48 h with 50 ng/ml (memory) and 1 ug/ml (effector) of plate-bound anti-CD3 plus 1 ug/ml of antiCD28 (both from BD PharmingenTM).