“…At the end of the experiment, mice were killed by cervical dislocation, spleens were removed and single cell suspensions were prepared by teasing the tissue apart. Erythrocytes were lysed by incubating the cells for 1 min in lysis buffer (155 mm NH 4 Cl, 10 mm KHCO 3 , 100 m EDTA-disodium, pH 7.4). After extensive washing splenocytes were passed through a 100-m cell strainer (Becton Dickinson Labware, Franklin Lakes, NJ, USA) to remove cell debris and were subsequently analyzed for cytokine production.…”